Metode HPLC Knauer PDF

Applications Journal
V7801
Contents
ENVIRONMENTAL APPLICATIONS 4
FOOD APPLICATIONS 27
PHARMACEUTICAL APPLICATIONS 59
CHIRAL APPLICATIONS 90
CHIRAL APPLICATIONS 127
SUBSTANCE AND KEYWORD INDEX 134
This Applications Journal was designed to provide you with detailed

information on the chromatographic conditions used with a variety of
HPLC applications. With our extensive line of HPLC products and
expert technical service we are positioned to fulfill all of your
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V 09/2008 3
Environmental Applications
Page
Determination of multiple Additives in cosmetics 5
Determination of Aldehydes and Ketones as DNPH derivates 6
Ion pair chromatography of Alkyl Sulphates and Alkyl Sulphosuccinates 7
Anion determination according to U.S. EPA Method 300.0 / 317.0 and 326.0
with suppressed conductivity 8
Determination of Anions with Ion pair chromatography and indirect UV detection 9
Determination of Carbamate Insecticides by HPLC with post-column
derivatization 10
Determination of monovalent Cations 11
Determination of divalent Cations 12
Separation of Explosives 13
Biomonitoring of Hydroxypyrene 14
Determination of Ketones as DNPH Derivates 15
Analysis of Nitro Polycyclic Aromatic Hydrocarbons 16
Separation of ortho-, nitro- para Nitroaniline 17
Fast Determination of PAH 18
Fast Separation of PAHs on a 2 mm ID column 19
Separation of Parabens 20
Chromatographic determination of Pesticides 21
Determination of Phenoxy acid herbicides 22
Determination of Priority Pollutant Phenols 23
Determination of non-ionic Surfactants using HPLC 24
Analysis of Surfactants in toothpastes by HPLC 25
Determination of aromatic Hydrocarbon types in middle distillates acc. to prEN
12916:2006 26
4 V 09/2008
1 Determination of multiple Additives in cosmetics
Method Matrix
HPLC cosmetics
Column: Eurospher 100-5 C18, 250 x 4 mm ID Order No. 25DE181ESJ
Phase: Eurospher 100-5 C18
Conditions: Eluent: Methanol / Water (35:65), adjusted to pH 2.5 with Phosphoric Acid
Gradient: 26 min linear gradient to Methanol / Phosphoric Acid (1000:1)
12 min final elution
Flow rate: 1.0 ml/min
Temperature: RT
Volume: 20 µl (50 µg/ml of each component)
Detection: UV at variable wavelengths (205 nm, 230 nm, 255 nm, 285 nm)
Substances: Benzyl nicotinate, Butyl p-hydroxybenzoate, 2,6-Di-tert.-butyl-4-methylphenol, 4-

Isopropyl-3-methylphenol, Methyl p-hydroxybenzoate, Monoammonium
glycyrrhizinate, Pantothenylethylether, Pyridoxine dioctanoate, Salicylic acid, Stearyl
glycyrrhetinate, Trichlorocarbanilide, Tocopheryl acetate
Keywords: Additives
Chromatogram:
3
1. Pantothenyl ethyl
2. Methyl p-
hydroxybenzoate
3. Salicylic acid
4. Benzyl nicotinate
5. 4-Isopropyl-3- 9
methylphenol 7 8
6. Butyl p- 12
hydroxybenzoate 10
7. Monoammonium
glycyrrhizinate 5
8. Trichlorocarbanilide 4
6
9. 2,6-Di-tert.-butyl-4- 2 11
methylphenol 1
10. Pyridoxine dioctanoate
11. Tocopheryl acetate
12. Stearyl glycyrrhetinate
0 10 20 30 40 min
V 09/2008 5
2 Determination of Aldehydes and Ketones as DNPH
derivates
Method Matrix
HPLC natural gas
Collection on a DNPH coated solid phase cartridge, elution with methanol
Conditions: Eluent: Methanol / Water (84:16)

Gradient: isocratic
Temperature: 30 °C
Volume: 10 µl
Detection: UV at 365 nm
Substances: DNPH derivates of Formaldehyde, Acetaldehyde, Acrolein, Propionaldehyde,

Acetone, Butyraldehyde, Methacrolein, Methyl ethyl ketone, Pentanone,
Cyclohexanone
Keywords: Aldehydes, Carbonyl compounds, Ketones
Chromatogram: 1
7
2
1. DNPH
2. Formaldehyde
3. Acetaldehyde
4. Acrolein
5. Propionaldehyde 9
6. Acetone
7. n-Butyraldehyde and
Methacrolein 3
8. Methyl ethyl ketone
6
8
9. 2-Pentanone and 4
Cyclohexanon 5
0 5 10 15 min
6 Environmental Applications
3 Ion pair chromatography of Alkyl Sulphates and
Alkyl Sulphosuccinates
Method Matrix
HPLC
Conditions: Eluent: 0.01 M aqueous Tetrabutylammonium Sulphate / Methanol (23:77),

adjusted to pH 3.0 with 0.01 M HCl
Gradient: isocratic
Temperature: ambient
Volume: 50 µl
Detection: Refractive Index
Substances: Octyl Hydrogen Sulphate, C10 Sulphosuccinate,

C12 Sulphosuccinate, C14 Sulphosuccinate
Keywords: Surfactants
Chromatogram:
1
1. Octyl hydrogen sulphate
2. C10 Sulphosuccinate
0 2 4 6 8 min
V 09/2008 7
4 Anion determination according to U.S. EPA Method
300.0 / 317.0 and 326.0 with suppressed conductivity
Method Matrix
HPLC
Column: Novosep A-2 Anion, 5 µm, 250 x 4 mm ID Order No. B92
Phase: Novosep A-2 Anion
Conditions: Eluent: 3.6 mM Sodium Carbonate (Na2CO3)

Gradient: isocratic
Temperature: 45 °C
50 µl (Chromatogram for a mixture of anions with concentration of
Volume:
0.25 ppm)
Detection: suppressed Conductivity (Alltech Model 650), Range 0.5 µS
Substances: Fluoride, Bromate, Chloride, Nitrite, Bromide, Nitrate, Phosphate, Sulfate
Keywords: Anions
Chromatogram: 1 4
1. Fluoride
2. n.n.
3. Bromate
4. Chloride
5. Nitrite
6. Bromide
7. Nitrate
8. Phosphate
9. Sulfate
7
3 5
6 9
8
2
0 5 10 15 20 25 min
5 Determination of Anions with Ion pair chromatography
and indirect UV detection
Method Matrix
HPLC
Column: Eurospher 100-5 C18, 250 x 4.6 mm ID Order No. 25EE181ESJ
Conditions: Eluent: 0.11 % Acetyl trimethylammonium p-toluenesulphonate +

0.002 M p-Toluenesulphonate in Acetonitrile / Water (39:61),
adjusted to pH 5.5 - 6 with Ammonium hydroxide
Volume: 10 µl
Detection: UV at 236 nm and 254 nm,

Chromatograms for a mixture of anions (1µg each, S.P. = system peak)
Substances: Cl-, ClO3-, SO42-, NO3-, I-, IO3-, SCN-, Br-
Keywords: Anions
Chromatogram:
6
1. IO3-
-
2. Cl
3. Br- 1
7
4. NO3-
5. ClO3-
6. I-
7. SCN-
8. SO42-
4 6
1
5 8
8
3 5 7
3
2
4
2
236 nm 254 nm
V 09/2008 9
6 Determination of Carbamate Insecticides by HPLC
with post-column derivatization
Method Matrix
HPLC
Conditions: Eluent: A: Water B: Methanol

Gradient: 0 – 0.5 isocratic, 82 % A
0.5 – 29 linear, 82 – 30 % A
29 – 31 isocratic, 100 % B, column rinsing
31 – 36 isocratic, 82 % A, re-equalibration
Temperature: 42 °C
Volume: 10 µl
two step reaction
Post-column
1 st Reactor (500 µl): 100 °C for first derivatization step with NaOH
derivatization
2nd Reactor (150 µl): ambient with OPA and 2-mercaptoethanol derivative
Detection: Fluorescence: excitation 330 nm, emission 465 nm
Substances: Aldicarb, Aldicarb sulphoxide, Aldicarb sulphone, BDMC, Carbaryl, Carbofuran, 3-

Hydroxycarbofuran, Methiocarb, Methomyl, 1-Naphtol, Oxamyl, Propoxur
Keywords: Carbamates, Insecticides
Chromatogram:
4
1. Aldicarb sulphoxide 9
2. Aldicarb sulphone
3. Oxamyl
4. Methomyl 2
5. 3-Hydroxycarbofuran 1
3 7
6. Aldicarb 6
7. Propoxur 5
8. Carbofuran 8
9. Carbaryl 10
10. 1- Naphtol 11
11. Methiocarb 12
12. BDMC
0 10 20
7 Determination of monovalent Cations
Method Matrix
HPLC
Column: PRP-X 200, 150 x 4.1 mm with Precolumn Order No. B89
Phase: PRP-X 200, cation exchange resin
Conditions: Eluent: 4 mM Nitric Acid in 30 % MeOH

Gradient: isocratic
Temperature: 35 °C
Volume: 20 µl
Detection: Conductivity (negative Polarity)
Substances: Ammonium, Lithium, Potassium, Sodium
Keywords: Cations, monovalent
Chromatogram:
1. Lithium (5.0 ppm)

2. Sodium (10.0 ppm) 3
2
3. Ammonium (10.0 ppm)
4. Potassium (10.0 ppm)
4
0 2 3 4 5 min
V 09/2008 11
8 Determination of divalent Cations
Method Matrix
HPLC
Column: PRP-X 200, 150 x 4.1 mm with Precolumn Order No. B89
Phase: PRP-X 200, cation exchange resin
Conditions: Eluent: 1.5 mM Ethylene Diammonium Dichloride

Gradient: isocratic
Temperature: 35 °C
Volume: 10 µl
Detection: Conductivity (negative Polarity)
Substances: Barium, Calcium, Magnesium, Strontium
Keywords: Cations, divalent
Chromatogram: 1
1. Magnesium (20.0 ppm)

2. Calcium (20.0 ppm)
3. Strontium (20.0 ppm)
4. Barium (20.0 ppm) 2
3
4
0 2 5 8 min
9 Separation of Explosives
Method Matrix
HPLC
Column: Ultra Sep ES EX, 250 x 3 mm ID Order No. I0034
Conditions: Eluent: A: Methanol B: Water

Gradient: linear, 25-60 % Methanol in 40 min,
5 min hold and equilibrate column
5 min with 25 % A for next injection
Temperature: 40 °C
Volume: 10 µl
Substances: 2-Amino-4,6-dinitrotoluene, 4-Amino-2,6-dinitrotoluene, 2-Amino-4-nitrotoluene, 2-

Amino-6-nitrotoluene, 1,3-Dinitrobenzene, 2,4-Dinitrotoluene, 2,6-Dinitrotoluene,
Hexogen, Nitrobenzene, 2-Nitrotoluene, 4-Nitrotoluene, 3-Nitrotoluene, Octogen,
Tetryl
Keywords: Explosives, EPA 8330
Chromatogram: 13
9
7
1. Octogen 1
2. Hexogen
3. 1,3,5-Trinitrobenzene
4. 2-Amino-6-nitrotoluene 3 6
5. 2-Amino-4-nitrotoluene
6. 1,3-Dinitrobenzene
7. Tetryl
8. Nitrobenzene
9. 2,4,6-Trinitroluene
10. 4-Amino-2,6-dinitrotoluene 12
11. 2-Amino-4,6-dinitrotoluene 10
8
12. 2,6-Dinitrotoluene
13. 2,4-Dinitrotoluene 2 4 5 14
14. 2-Nitrotoluene 11 16
15. 4-Nitrotoluene
16. 3-Nitrotoluene
15
0 5 10 15 20 25 30 35 min
V 09/2008 13
10 Biomonitoring of Hydroxypyrene
Method Matrix
HPLC
Column: Eurospher 100-5 C18, 250 x 4 mm Order No. 25DE181ESJ
Conditions: Eluent: A: ACN B: H2O

Gradient: linear 60 % A in 28 min to 100 % A, 5 min hold, equilibrate column
10 min with 60 % A for next injection
Temperature: 30 °C
Volume: 10 µl
Substances: Hydroxypyrene, Biomarker of PAH contamination
Keywords: Hydroxypyrene
Chromatogram: 1
1. Hydroxypyrene
0 5 10 15 20
11 Determination of Ketones as DNPH Derivates
Method Matrix
HPLC
Column: ProntoSIL 120-3 C18 AQ, 250 x 3 mm ID Order No. 15DF080PSJ
Phase: ProntoSIL 120-3 C18 AQ
Conditions: Eluent: A: Water B: Acetonitrile

Gradient: 10 – 100 % B in 18 min
Temperature: 30 °C
Volume: 2 µl
Substances: DNPH derivates of Formaldehyde, Acetaldehyde, Acrolein, Propionaldehyde,

Acetone, Butyraldehyde, Methacrolein, Methyl ethyl ketone, Pentanone,
Cyclohexanone
Keywords: Aldehydes, Carbonyl compounds, Ketones
Chromatogram: 1
1. Tetramethylketone
2
2. 2-Pentanone
3. 2-Hexanone
4. 2-Heptanone
5. 2-Octanone
6. 2-Nonanone
7. 2-Decanone
3
8. 2-Undecanone
9. 2-Dodecanone
10. 2-Tridecanone
11. 2-Hexadecanone 4
10
5 8 11
6 7 9
0 5 10 15 min
V 09/2008 15
12 Analysis of Nitro Polycyclic Aromatic Hydrocarbons
Method Matrix
HPLC
Column: ProntoSIL 120-3 Phenyl, 125 x 4 mm ID Order No. 12DF050PSG
Phase: ProntoSIL 120-3 Phenyl
Conditions: Eluent: A: Methanol / Water (60:40) B: Methanol

Gradient: 100 % A for 13 min
0 – 100 % B in 29 min
Temperature: 25 °C
Volume: 2 µl
Detection: Fluorescence (wavelength program)
Substances: 2-Nitro-Naphthalene, 2-Nitro-Fluorene, 9-Nitro-Phenanthrene,

1-Nitro-Pyrene, 6-Nitro-Chrysene, 6-Nitro-Benzo(a)pyrene
Keywords: Nitro-PAH
Chromatogram: 2
4
1. Nitro-Naphthalene 5
2. Nitro-Fluorene
3. Nitro-Phenanthrene
4. Nitro-Pyrene
5. Nitro-Chrysene
6. Nitro-Benzo(a)pyrene
6
1
0 10 20 30 min
13 Separation of ortho-, nitro- para Nitroaniline
Method Matrix
HPLC
Column: Eurospher 100-5 Si, 120 x 4 mm ID Order No. 11DE000ESJ
Phase: Eurospher 100-5 Si
Conditions: Eluent: A: Heptan B: Ethanol

Gradient: isocratic (85 % A / 15 % B)
Temperature: 30 °C
Volume: 10 µl
Substances: 2-Nitroaniline, 3-Nitroaniline, 4-Nitroaniline
Keywords: Nitroaniline
Chromatogram: 2
1. 2-Nitroaniline
2. 3-Nitroaniline
3. 4-Nitroaniline
3
1
0 2 4 6 8 min
V 09/2008 17
14 Fast Determination of PAH
Method Matrix
HPLC
Column: UltraSep ES PAH QC, 60 x 2 mm ID Order No. I0019
Phase: UltraSep ES PAH QC
Conditions: Eluent: A: Water / B: Acetonitrile

Gradient: 55 % B at start; 55 % B 0-5 min; 100 % B from 5-12 min, 8 min
hold
Temperature: 30 °C
Detection: UV at 254 nm and/or Fluorescence (wavelength program)
Substances: Naphthalene, Acenaphthylene, Acenaphthene, Fluorene, Phenanthrene, Anthracene,

Fluoranthene, Pyrene, Benzo(a)anthracene, Chrysene, Benzo(b)fluoranthene,
Benzo(k)fluoranthene, Benzo(a)pyrene, Dibenzo(a,h)anthracene,
Benzo(g,h,i)perylene, Indeno(1,2,3-cd)pyren, 1-Methylnaphthalene, 2-
Methylnaphthalene, Benzo(e)pyrene
Keywords: Polyaromatic hydrocarbons
Chromatogram:
1. Naphthalene 11
2. Acenaphthylene
3. Acenaphthene 17
4. Fluorene
5. Phenanthrene 18
1 6 10
6. Anthracene 2
7. Fluoranthene
8. Pyrene 9
9. Benzo(a)anthracene 5 19
10. Chrysene 4 13
11. Benzo(b)fluoranthene 7
12
12. Benzo(k)fluoranthene 16
13. Benzo(a)pyrene 3 8 15
14. Dibenzo(a,h)anthracene 14
15. Benzo(g,h,i)perylene
16. Indeno(1,2,3-cd)pyrene
17. 1-Methylnaphthalene
18. 2-Methylnaphthalene
19. Benzo(e)pyrene min
15 Fast Separation of PAHs on a 2 mm ID column
Method Matrix
HPLC
Column: Eurospher 100-3 C18, 250 x 2 mm Order No. 25BE181ESG
Conditions: Eluent: A: MeOH / Water (80:20) B: ACN / THF (93:7)

Gradient: linear 0-100 % B in 15 min, 5 min hold;
equilibrate column 10 min with 100 % A for next injection
Temperature: 30 °C
Volume: 5 µl
Detection: Fluorescence, wavelength program (excitation / emission):

1. 275/350 nm, 2. 260/390 nm, 3. 260/420 nm, 4. 290/430 nm, 5. 300/500 nm
UV at 254 nm
Substances:
Naphthalene, Acenaphthylene, Acenaphthene, Fluorene, Phenanthrene,
Anthracene, Fluoranthene, Pyrene, Benzo(a)anthracene, Chrysene,
Benzo(b)fluoranthene, Benzo(k)fluoranthene, Benzo(a)pyrene,
Dibenzo(a,h)anthracene, Benzo(g,h,i)perylene, Indeno(1,2,3-cd)pyren
Keywords: 16 PAH´s according to EPA (550.1 / 610 / 8310)
Chromatogram: 7
1. Naphthalene 5
2. Acenaphthylene 6 9
3. Acenaphthene
4. Fluorene
5. Phenanthrene 4
6. Anthracene
7. Fluoranthene 10
14
8. Pyrene
9. Benzo(a)anthracene 12
10. Chrysene 8 11
3
11. Benzo(b)fluoranthene
1 13
12. Benzo(k)fluoranthene
13. Benzo(a)pyrene 2
14. Dibenzo(a,h)anthracene
15. Benzo(g,h,i)perylene
16. Indeno(1,2,3-cd)pyrene 15
16
0 10 20 min
V 09/2008 19
16 Separation of Parabens
Method Matrix
HPLC
Column: ProntoSIL 120-3 Phenyl, 125 x 4 mm ID Order No. 12DF050PSG
Phase: ProntoSIL 120-3 Phenyl
Conditions: Eluent: 20 mM Potassium Hydrogen Phosphate (KHPO4) / Acetonitrile (50:50)

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: Methylparabene, Ethylparabene, Propylparabene, Butylparabene
Keywords: Parabenes
Chromatogram: 2
1. Methylparabene 1
2. Ethylparabene
3. Propylparabene
4. Butylparabene
3
4
1 3 5 min
17 Chromatographic determination of Pesticides
Method Matrix
HPLC
Column: UltraSep ES PEST, 250 x 3 mm ID Order No. I0025
Phase: UltraSep ES PEST
Conditions: Eluent: ACN / Water

Gradient: from 20 to 26 % ACN within 5 min,
26 to 30 % ACN within 19 min
30 to 60 % within 12 min
Temperature: 30 °C
Volume: 10 µl
Detection: UV at 215 nm and 230 nm
Substances: Atrazine, Chlortoluron, Cyanazin, Desethylatrazine, Diuron, Hexazinon,

Isoproturon, Linuron, Metoxuron, Methabenzthiazuron, Metolachlor,
Metobromuron, Metazachlor, Monolinuron, Sebuthylazin, Simazin,
Terbuthylazine
Keywords: Pesticides
Chromatogram: 15
1. Desethylatrazin
2. Hexazinon 1
3. Metoxuron
4. Simazin 4
5. Cyanazin
6. Methabenzthiazuron
7. Atrazine
8. Chlortoluron
5 14
9. Monolinuron 7
10. Isoproturon 6
11. Metobromuron
12. Diuron
13. Metazachlor 3
14. Sebuthylazin 10
15. T erbuthylazine 2 9 17
8
16. Linuron 11 16
17. Metolachlor 12
13
5 10 15 20 25 30 min
V 09/2008 21
18 Determination of Phenoxy acid herbicides
Method Matrix
HPLC
Column: UltraSep ES PHENOXYCARB, 250 x 2 mm ID Order No. I0202
Phase: UltraSep ES PHENOXYCARB
Conditions: Eluent: A: Water (TFA adj. pH 2.5) B: Acetonitrile

Gradient: 0 % B at start; 0-20 % B in 2 min; 20-35 % B in 28 min; 35-100%
B in 15 min; 5 min hold
Temperature: 60 °C
Substances: Clopyralid; Quinmerac ; Nicosulfurone ; Dicamba; Fluroxypyr; Mesotrione;

Sulcotrione; Bentazone; Bromoxynil; DNOC (4,6-Dinitro-o-cresol); 2,4-D (2,4-
Dichlorophenoxy acetic acid); MCPA (4-Chloro-o-tolyloxy acetic acid); Ioxynil ;
Dichlorprop; Mecoprop; 2,4,5-T (2,4,5-Trichlorophenoxy acetic acid); 2,4-DB (4-(2,4-
Dichlorophenoxy)butyric acid); MCPB (4-(4-Chloro-o-tolyloxy)butyric acid); Fluazifop-
p; Fenoxyprop-p; Haloxyfop; Dinoterb
Keywords: Phenoxy acid herbicides
1. Clopyralid
2. Quinmerac
3. Nicosulfurone
4. Dicamba
5. Fluroxypyr
6. Mesotrione
7. Sulcotrione
8. Bentazone
9. Bromoxynil 2
10. DNOC (4,6-Dinitro-o-cresol)
11. 2,4-D (2,4 –Dichlorophenoxy
acetic acid)
12. MCPA (4-Chlor-o-tolyloxy 21
acetic acid) 9 19
20
13. Ioxynil
14. Dichlorprop
22
15. Mecoprop 1 18
5 6
16. 2,4,5-T (2,4,5- 4 8 17
13
Trichlorophenoxy acetic acid) 11
7
17. 2,4-DB (4-(2,4- 12 15
14
10
Dichlorophenoxy)butyric acid) 3 16
18. MCPB (4-(4-Chlor-o-tolyloxy)

butyric acid)
19. Fluazifop-p
20. Fenoxyprop-p
21. Haloxyfop
22. Dinoterb 0 10 20 30 40 min
19 Determination of Priority Pollutant Phenols
Method Matrix
HPLC
Conditions: Eluent: A: MeOH + 1 % Acetic Acid B: Water plus 1 % Acetic Acid

Gradient: 30 % A in 9 min to 40 % A
40 % A in 7 min to 100 % A, 6 min hold
Temperature: 40 °C
Volume: 5 µl
Substances: 2-Chlorophenol, 4-Chloro-3-methylphenol, 2,4-Dichlorophenol,

2,3-Dimethylphenol, 2,4-Dinitrophenol, 2-Methyl-4,6-dinitrophenol,
2-Nitrophenol, 4-Nitrophenol, Pentachlorphenol, Phenol,
2,4,6-Trichlorphenol
Keywords: Phenols
Chromatogram:
9
1. Phenol
2. 4-Nitrophenol
3. 2,4-Dinitrophenol 11
4. 2-Chlorophenol 7
5. 2-Nitrophenol 10
6. 2,3-Dimethylphenol 2
7. 2-Methyl-4,6-dinitrophenol 8
8. 4-Chloro-3-methylphenol 6
9. 2,4-Dichlorphenol 1
4
10. 2,4,6-Trichlorphenol 3 5
11. Pentachlorphenol
0 5 10 15 20 min
V 09/2008 23
20 Determination of non-ionic Surfactants using HPLC
Method Matrix
HPLC
Conditions: Eluent: MeOH / Water (90:10)

Temperature: 25 °C
Volume: 20 µl
Substances: Polyethylene glycol, Polyoxyethylene sorbitane monolaurate, Nonylphenol

polyglycol ether, Laurylalcohol, Laurylalcohol polyglycol ether, Oleic acid
diethanolamide, Oleic amide polyglycol ether, Myristylalcohol, Myristyl alcohol
polyglycol ether, Palmitic acid polyglycol ester, Polyoxyethylene glycerol
monostearate, Stearic acid polyglycol ester
Chromatogram: 1
1. Polyethylene glycols
2. Polyoxyethylene sorbitane monolaurate
3. Nonylphenol polyglycol ether
4. Lauryl alcohol 3
5. Lauryl alcohol polyglycol ether 6
6. Oleic acid diethanolamide
7. Oleic amide polyglycol ether 5
8. Myristyl alcohol
9. Myristyl alcohol polyglycol ether 8
10. Palmitic acid polyglycol ester 4 7
11. Polyoxyethylene glycerol monostearate
12. Stearic acid polyglycol ester. 2 10
12
9 11
0 5 10 15 min
21 Analysis of Surfactants in toothpastes by HPLC
Method Matrix
HPLC toothpaste
Conditions: Eluent: MeOH / Water (90:10) + 0.25 M NaClO4

Gradient: isocratic
Temperature: 25 °C
Volume: 20 µl
Substances: Sodium soap (C8 - C18),

Sulpho fatty acid-α-methyl esters (C10 - C18)
Chromatogram:
Sodium soaps:
1. (C8) 2
2. (C10)
3. (C12)
4. (C14)
5. (C16)
6. (C18 =)
7. (C18)
α-sulpho fatty acid methyl esters: 6

1. (C10)
2. (C12) 3 5
3. (C14) 3
4. (C16)
5. (C18)
4 7
1
4 5
12
5 10 15 min 5 10 15
V 09/2008 25
22 Determination of aromatic Hydrocarbon types in middle
distillates acc. to prEN 12916:2006
Method Matrix diesel fuel

HPLC
Sample preparation: dilution with n-heptane
Column: Ultrasep ES AP, 5 µm, 250 x 4 mm ID Order No. 25DE450USJ
Phase: Ultrasep ES AP 5 µm
Conditions: Eluent: n-Heptane (dried with molecular sieve)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Detection: RI
Substances: MAH: phenyldodecane; 1,2 dimethylbenzene; hexamethylbenzene

DAH: naphthalene, fluorene
T-AH: dibenzothiphene; phenanthrene, 9-methylanthracene
Keywords: MAH (mono-aromatic compounds); DAH (di-aromatic compounds);

T+AH (tri+-aromatic compounds)
Chromatogram: Cyclohexane
1
Retention time standard
1. Cyclohexane MAH 20.14 % [m/m]
2. Phenyldodecane Sample
3. 1,2 Dimethylbenzene 11
4. Hexamethylbenzene
5. Naphthalene 3
Total aromatic hydrocarbons: 22.59 % [m/
8
6. Dibenzothiophene
7. 9-Methylanthracene
Calibration standard 1
8. 1,2 Dimethylbenzene
9. Fluorene DAH 2.33 % [m/m]
12
10. Phenanthrene 9
T+AH 0.11 % [m/m]

Sample 13
11. MAH 20.14% [m/m]
5
12. DAH 2.33% [m/m] 4
0 5 10 15 min
13. T+AH 0.11% [m/m] 2 7

Total aromatic hydrocarbens: 6
10
22.59 % [m/m]
0 5 10 15 min 0 5 10 15 min
Retention time standard Calibration standard
26
Food Applications
Page
Separation of Additives in soft drinks 28
Determination of Aflatoxins by HPLC with post column derivatization 29
Simultaneous determination of Alcohols, Carbohydrates and Organic Acids 30
Determination of Alcohols, Carbohydrates and Organic Acids II 31
Separation of Amines in wine with precolumn derivatization (DANSYL) 32
HPLC determination of Amines in wine with precolumn derivatization (OPA) 33
Determination of Amino Acids in beer with precolumn derivatization (OPA) 34
Inorganic and Organic Anions in beer 35
Anthocyanins in red and rose wine 36
Determination of Biogenic Amines in food products by HPLC in combination
with ninhydrin derivatization 37
Separation of β-Carotene isomers 38
Determination of Diketopiperazine in soft drinks 39
Determination of Fatty acids 40
Analysis of Flavonoids in fruit juice 41
Stilbene and Flavonoids in red wine samples 42
Analysis of Flavonoid Glycosides 43
Separation of Methylfurfural and Hydroxyfurfural in beer 44
Determination of Mono- and Disaccharides 45
Determination of Deoxynivalenol (DON) 46
Determination of Nivalenol (NIV) and Deoxynivalenol (DON) with postcolumn
derivatisation 47
Analysis of Fusarium Mycotoxines 48
Ion chromatography for determination of NO2- and NO3- in beverages 49
Determination of Organic Acids 50
Determination of Organic Acids II 51
Determination of Patulin in apple juice 52
Separation of Preservatives 53
Determination of various Sterols 54
Separation of water soluble Vitamins 55
Separation of Tocopherols and Vitamin D2 56
Separation of fat soluble Vitamins 57
Fat soluble Vitamins by HPLC with electrochemical detection 58
27
1 Separation of Additives in soft drinks
Method Matrix
HPLC
Column: ProntoSIL 120-5 C8 SH, 150 x 4 mm ID Order No. 15DF080PSJ
Phase: ProntoSIL 120-5 C8 SH
Conditions: Eluent: 20 mM KH2PO4 (pH 3) / Acetonitrile (5:1)

Gradient: isocratic
Temperature: 30 °C
Volume: 10 µl
Substances: Acesulfam K, Ascorbic acid, Aspartame, Benzoic acid, Caffeine, Saccharin, Sorbic
acid, Quinine, Vanilin
Keywords: Sweeteners, Additives
Chromatogram:
2
1. Ascorbic acid
2. Acesulfam K
3. Saccharin
4. Caffeine
5. Aspartame
6. Quinine 4
7. Vanilin
8. Sorbic acid
9. Benzoic acid
5
1
3
8
6 7
9
0 5 10 min
28 Food Applications
2 Determination of Aflatoxins by HPLC with
post column derivatization
Method Matrix
HPLC food
Column: UltraSep ES TOX 1 / UltraSep ES TOX 2; 125 x 3 mm ID Order No. I0200 / I0201
Phase: UltraSep ES TOX 1 / UltraSep ES TOX 2
Conditions: Eluent: Methanol / Water (60 / 40) / (35 / 65)

Gradient: isocratic
Volume: 20 µl / 3 µl
Detection: postcolumn derivatization with 1.3 mM bromine solution at a flow rate of 0.18 ml/min,
fluorescence: excitation 375 nm, emission 455 nm to get a detection limit of 0.5 ppm
Substances: Aflatoxin B1, Aflatoxin B2, Aflatoxin G1, Aflatoxin G2
Keywords: Aflatoxins
Chromatogram: UltraSep ES TOX 1 UltraSep ES TOX 2

1. Aflatoxin G2
2. Aflatoxin G1
3. Aflatoxin B2
4. Aflatoxin B1
V 09/2008 29
3 Simultaneous determination of Alcohols,
Carbohydrates and Organic Acids
Method Matrix
HPLC beverage
Column: Eurokat H 10 µm, 300 x 8 mm ID Order No. 30GX340EKN
Phase: Eurokat H 10 µm
Conditions: Eluent: 0.01 M Sulfuric acid

Gradient: isocratic
Temperature: 80 °C
Volume: 10 µl
Substances: Maltose, Glucose, Succinic acid, Lactic acid, Formic acid, Acetic acid, Methanol,
Ethanol
Keywords: Carbohydrates, Organic acids, Alcohols
Chromatogram: 3
1. Higher carbohydrates
2. Maltose
3. Glucose
4. Succinic acid
5. Lactic acid 2
6. Formic acid
7. Acetic acid
8. Methanol
5
9. Ethanol
4
7
1 6
0 10 20 30 min
4 Determination of Alcohols, Carbohydrates and
Organic Acids II
Method Matrix
HPLC
Column: Eurokat H 10 µm, 300 x 8 mm ID Order No. 30GX340EKN
Phase: Eurokat H 10 µm
Conditions: Eluent: 0.00125 N Sulfuric Acid (pH 2)

Gradient: isocratic
Temperature: 90 °C
Volume: 10 µl
Substances: Acetic acid, Butandiol, Citric acid, Ethanol, Fructose, Glucose, Lactic acid, Malic acid,
Succinic acid, Tartaric acid
Keywords: Organic acids
Chromatogram: 3
1. Citric acid
2. Tartaric acid
3. Glucose
4. Malic acid
5. Fructose
6. Sucinic acid
7. Lactic acid
8. Acetic acid
9. Butandiol 1 4
10. Ethanol
2
5
6
7 8
10
9
0 5 10 15 20 25 30 35
V 09/2008 31
5 Separation of Amines in wine with precolumn
derivatization (DANSYL)
Method Matrix
HPLC wine
Conditions: Eluent: ACN / Water (30:70)

Gradient: over 40 min to 60 % ACN and 15 % Ethanol in water, 10 min hold
Volume: 20 µl
Substances: Ethanolamine, Ammonia, Methylamine, Serotonin, Ethylamine, Morpholine,

Isopropylamine, Dimethylamine, Propylamine, Pyrrolidine, Pyrrolidone, Octopamine,
Indole, Butylamine, Diethylamine, Phenethylamine, Methylbutylamine, Isopentylamine,
Pentylamine, Piperidine, Diaminopropane, Diaminobutane, Hexylamine, Diaminopentane,
Histamine
Keywords: Amines
Chromatogram:
10-13
1. Ethanolamine,
22
2. Ammonia,
3. Methylamine,
4. Serotonin, 20,21
5. Ethylamine,
6. Morpholine, 7,8
7. Isopropylamine,
8. Dimethylamine, 19 23,24
9. Propylamine
14 17,18
10. Pyrrolidine, 9 15 16
11. 2-Pyrrolidone, 5
12. Octopamine,
13. Indole 3
14. Butylamine,
6
15. Diethylamine,
1
16. Phenethylamine,
25
17. 2-Methylbutylamine, 2 4
18. Isopentylamine,
19. Pentylamine,
20. Piperidine,
21. 1,3-Diaminopropane,
0
22. 1,4-Diaminobutane, 10 20 30 40 50 min
23. Hexylamine,
24. 1,5-Diaminopentane,
25. Histamine
6 HPLC determination of Amines in wine with
precolumn derivatization (OPA)
Method Matrix
HPLC wine
Conditions: Eluent: 0.08 M Acetic Acid / Acetonitrile (70:30) 3 min

Gradient: in 6.5 min to 65 % Acetonitrile, in 11 min to 72 % Acetonitrile,
in 1 min to 80 % Acetonitrile, 3 min hold
Volume: 20 µl
Detection: Fluorescence: ex 230 nm, em 440 nm
Substances: Cadaverine, Heptylamine, Histamine, Isoamylamine, Pentyl-i-amine, Phenethylamine,

Putrescine
Keywords: Amines
Chromatogram:
1. Histamine 3
2. Tyramine 2
3. Putrescine
4. Phenethylamine
5. Cadaverine
6. Isoamylamine
7. internal Standard IS 7
456
0 5 10 15 20 min
V 09/2008 33
7 Determination of Amino Acids in beer with
precolumn derivatization (OPA)
Method Matrix
HPLC beer
Column: OPA column B801, 300 x 4 mm ID Order No. B801
Phase: OPA Column
Conditions: Eluent: A: 0.05 M Acetate / Methanol (81:19) pH 7.2

B: 0.05 M Acetate / Methanol (25:75)
Gradient: 28 min 100 % A, in 7 min to 70 % A,
in 15 min to 100 % B (5 min hold), in 5 min 100 % A
Temperature: 40 °C
Volume: 10 µl
Substances: Aspartic acid, Glutamic acid, Asparagin, Serine, Glutamin, Histidine, Glycine,
Threonine, Arginin, Alanine, Tyrosine, Methionine, Valine, Phenylalanine, Isoleucine,
Leucine, Lysine
Keywords: Amines
Chromatogram: 14
1. ASP 9 11 13
2. GLU
3. ASN
4. SER
5. GLN
6. HIS 16
7. GLY
8. THR
9. ARG
7
10. ALA 2 10 12
11. TYR 15
12. MET
13. VAL
14. PHE
15. ILE 17
16. LEU
17. LYS 6
1
5
34 8
0 10 20 30 40 50 60 min
8 Inorganic and Organic Anions in beer
Method Matrix
HPLC beer
Column: Allsep Anion, 100 x 4.6 mm ID Order No. B90
Phase: Allsep Anion
Conditions: Eluent: 0.85 mM Sodium Hydrogen Carbonate (NaHCO3)

Gradient: isocratic
Temperature: 35 °C
Volume: 100 µl
Detection: suppressed Conductivity (Alltech Model 650), Range 10 µS
Substances: Chloride, Nitrate, Phosphate, Sulfate, Oxalate
Keywords: Anions, inorganic and organic
Chromatogram: 1
Chromatogram of beer sample
(dilution 1:10)
1. Chloride
2. Nitrate
3. Phosphate
4. Sulfate
5. Oxalate
2
5
5 10 15 min
V 09/2008 35
9 Anthocyanins in red and rose wine
Method Matrix
HPLC wine
ProntoSIL 120-3 C18 SH, 250 x 4 mm

Column: Order No. 25DF180PSG
ID
Conditions: Eluent: A: Water / Formic Acid / Acetonitrile (87:10:3)

B: Water / Formic Acid / Acetonitrile (pH 1.3)
Gradient: 0 min 94 % A 6 % B, 15 min 70 % A 30 % B,
30 min 50 % A 50 % B, 35 min 40 % A 60 % B,
41 min 94 % A 6 % B (2 min hold)
Temperature: 40 °C
Volume: 20 µl
Substances: Group 1: nonacylated anthocyanidin-3-glycosides

Group 2: acylated anthocyanidin-3-glycosides
Group 3: coumarylated anthocyanidin-3-glycosides
Keywords: Anthocyanins
Chromatogram:
Group 1
1. Delphinidol-3-glucosid
2. Cyanidol-3-glucosid 5
3. Petunidol-3-glucosid
4. Peonidol-3-glucosid
5. Malvidol-3-glucosid
Group 2
6. Peonidol-3-acetylglucosid
7. Malvidol-3-acetylglucosid
Group 3
8. Peonidol-3-coumarylglucosid
9. Malvidol-3-coumarylglucosid
4
3
1 2 67 89
0 5 10 15 20 25 30 35 40 45 min
10 Determination of Biogenic Amines in food products by
HPLC in combination with ninhydrin derivatization
Method Matrix
HPLC Food
Conditions: Eluent: 16 g Ninhydrin and 1.2 g Hydrindantin, dissolved in 322 ml Dimethyl

sulphoxide (DMSO) by sonication, plus 350 ml 2.8 M Sodium acetate
buffer pH 5.0, mixed with a solution of 2 g Sodium dodecyl sulphate in
618 ml DMSO - 710 ml Water
Gradient: isocratic
Volume: 10 µl
Detection: UV-VIS at 546 nm
Substances: Tyramine, Histamine, Putrescine, Cadaverine, Tryptamine, Phenylethylamine
Keywords: Amines, Histamine
Chromatogram:
3
1. Tyramine
2. Histamine
1
3. Putrescine
4. Cadaverine 4
5. Tryptamine
6. Phenylethylamine 2
6
5
0 10 20 30 40 50 60 70 min
V 09/2008 37
11 Separation of β-Carotene isomers
Method Matrix
HPLC
Column: ProntoSIL 200-3 C30, 250 x 4.6 mm ID Order No. 25EH300PSG
Phase: ProntoSIL 200-3 C30
Conditions: Eluent: MeOH / TBME (80:20)

Gradient: isocratic
Temperature: 25 °C
Volume: 5 µl
Substances: β-Carotene
Keywords: Vitamins, fat soluble
Chromatogram:
1
1. all-trans β-Carotene
2. 9-cis β-Carotene
0 5 10 15 20 25 min
12 Determination of Diketopiperazine in soft drinks
Method Matrix
HPLC soft drink
Conditions: Eluent: 10 mM KH2PO4 / ACN (85:15), adjusted to pH 4.0

Gradient: isocratic
Temperature: 30 °C
Volume: 10 µl
Substances: Diketopiperazine, Caffeine, Aspartame, Benzoate
Keywords: Additives, Sweeteners
Chromatogram:
1. DKP = diketopiperazine
2. Caf = caffeine 2 3
3. APM = aspartame 4
4. BA = Na benzoate
0 10 20 30 min
V 09/2008 39
13 Determination of Fatty acids
Method Matrix
HPLC
Column: UltraSep ES FS, 250 x 3 mm ID Order No. I0046
Phase: UltraSep ES FS
Conditions: Eluent: A: 10 mM Phosphoric acid pH 2.9 (NaOH)

B: Acetonitrile
Gradient: 70 % B at start; 70-98 % B in 30 min
Temperature: 30 °C
Substances: Capric acid; Lauric acid; Myristic acid; Palmitinic acid; Elaidinic acid; Stearinic acid;
Arachidonic acid
Keywords: Fatty acids
Chromatogram:
2
1. Capric acid
2. Lauric acid
3. Myristic acid
4. Palmitinic acid
5. Elaidinic acid
6. Stearinic acid 3
7. Arachidonic acid
1 6
4 7
0 5 10 15 20 min
14 Analysis of Flavonoids in fruit juice
Method Matrix
HPLC Beverage (orange juice sample)
Conditions: Eluent: 5 mM Ammonium acetate / ACN (75:25)

(pH 4,45; adjusted with conc. acetic acid)
Gradient: isocratic
Temperature: 40 °C
Volume: 5 µl
Substances: Naringin, Hesperidin
Keywords: Flavonoids
Chromatogram:
1. Naringin (28.03 ppm)

2. Hesperidin (79.25 ppm)
0 5 9 min
V 09/2008 41
15 Stilbene and Flavonoids in red wine samples
Method Matrix
HPLC wine
Conditions: Eluent: A: MeOH / H2O (50:50) (pH 3 with TFA)

B: MeOH / H2O (80:20) (pH 3 with TFA)
C: MeOH / H2O (20:80) (pH 3 with TFA)
Gradient: 0 – 5 min 90 % A 10 % C Æ 100 % A
5 – 8 min 100 % A Æ 70 % A 30 % B
8 - 12 min 70 % A 30 % B Æ 20 % A 80 % B
12 - 14 min 20 % A 80 % BÆ 100 % B
14 - 16 min 100 % B isocratic
Temperature: 40 °C
Volume: 20 µl
Detection: UV at different wavelength

(Resveratrol: 305 nm, Myrcetine: 254/365nm, Quercetine: 365 nm, Caempferol:
265/365 nm)
Substances: trans-Resveratrol, cis-Resveratrol, Myrcetine, Quercetine, Caempferol
Keywords: Stilbene, Flavonoids
Chromatogram: 5
1. trans Resveratrol 1
2. Myrcetine
3. cis Resveratrol
4. Quercetine
5. Caempferol
4
3
2
0 5 10 15 20 min
16 Analysis of Flavonoid Glycosides
Method Matrix
HPLC
Conditions: Eluent: ACN / H2O, adjusted to pH 2.5; with Phosphate buffer

Gradient: 17 - 23 % ACN over 35 min
Temperature: 25 °C
Volume: 10 µl
Substances: Eriocitrin, Eriodictyol 7-O-glucoside, Luteolin 7-O-rutinoside, Narirutin, Hesperidin,

Isorhoifolin, Rosmarinic acid, Diosmin
Keywords: Flavonoid Glycosides
Chromatogram:
1
1. Eriocitrin
2. Eriodictyol 7-O-glucoside
3. Luteolin 7-O-rutinoside
4. Narirutin
5. Hesperidin 7
6. Isorhoifolin 3
7. Rosmarinic acid
8. Diosmin
X1-4 unknown 5
4 8
2 x3
x1x2 x4
0 5 10 15 20 25 30 min
V 09/2008 43
17 Separation of Methylfurfural and Hydroxyfurfural in beer
Method Matrix
HPLC beer
Conditions: Eluent: H2O / ACN (90:10)

Gradient: isocratic
Temperature: 30 °C
Volume: 20 µl
Substances: Furfural, Hydroxymethylfurfural (HMF)
Keywords: HMF
Chromatogram: FURFURAL
HMF
HMF
FURFURAL
0 5 10 min 0 5 10 min
18 Determination of Mono- and Disaccharides
Method Matrix
HPLC
Column: Eurospher 100-5 NH2, 125 x 4 mm ID Order No. 12DE190ESJ
Phase: Eurospher 100-5 NH2

Gradient: isocratic
Temperature: 25 °C
Volume: 5 µl
Substances: Fructose, Glucose, Lactose, Maltose, Saccharose
Keywords: Saccharides
Chromatogram: 1
1. Fructose
2. Glucose
3. Saccharose
4. Maltose
5. Lactose
3
2
4 5
0 5 10 15 min
V 09/2008 45
19 Determination of Deoxynivalenol (DON)
Method Matrix
HPLC
Column: Eurospher 100-5 C18, 150 x 4 mm ID Order No. 15EE181ESJ
Conditions: Eluent: Water / Acetonitrile (90:10)

Gradient: isocratic
Temperature: 30 °C
Volume: 20 µl
Substances: Deoxynivalenol (DON)
Keywords: Mycotoxins, Trichotecenes
Chromatogram:
a.)
a) blank wheat sample

b) spiked wheat sample
0 5 10 min
b.)
1
0 5 10 min
20 Determination of Nivalenol (NIV) and Deoxynivalenol
(DON) with postcolumn derivatisation
Method Matrix
HPLC fungus
Column: ProntoSIL 120-5 C18 AQ, 250 x 3 mm ID Order No. 25CF184PSJ
Conditions: Eluent: A: ACN B: 0.01 M Acetic Acid

Gradient: 0-16 min 10 % A
16-22 min 90 % A
22-40 min 10 % A
Temperature: 25 °C
Volume: up to 200 µl
2 channel postcolumn derivatization, flow rate of 0.25 ml/min, temperature of 115 °C
Reactor Coil 1 : 1.2 ml, 0.15 M sodium hydroxide
Reactor Coil 2 : 1.6 ml, 0.03 M methyl acetoacetate and 2 M ammonium acetate
Detection: Fluorescence: ex at 360 nm, em at 470 nm
Substances: Deoxynivalenol (DON), Nivalenol (NIV)
Keywords: Mycotoxins, Trichothecens
Chromatogram:
1. Nivalenol (NIV)
2. Deoxynivalenol (DON)
1
2
0 5 10 15 min
V 09/2008 47
21 Analysis of Fusarium Mycotoxines
Method Matrix
HPLC
Conditions: Eluent: A: ACN B: Water / ACN (90:10)

Gradient: 0 – 28 min 8%A
28 – 32 min 20 % A
32 – 36 min 100 % A
36 – 40 min 99 % A
40 – 45 min 8%A
Flow rate: 0 – 7 min 0.65 ml/min
7 – 40 min 0.75 ml/min
40 – 45 min 0.65 ml/min
Temperature: 25 °C
Volume: 10 µl
Detection: DAD at 220 and 230 nm
Substances: Nivalenol, Deoxynivalenol (DON), 15-o-Acetyl-4-deoxynivalenol, 3-

Acetyldeoxynivalenol
Keywords: Fusarium Mycotoxines
Chromatogram: 2
1. Nivalenol (2.8 ppm)
2. Deoxynivalenol (2.9 ppm)
3. 15-o-Acetyl-4-deoxynivalenol (3.0 ppm) 1
4. 3-Acetyldeoxynivalenol (2.9 ppm)
3 4
0 5 10 15 min
22 Ion chromatography for determination of NO2- and NO3-
in beverages
Method Matrix
HPLC
Column: Novosep A-2 Anion, 250 x 4 mm ID Order No. B92
Conditions: Eluent: 3.6 mM Sodium carbonate (Na2CO3)

Gradient: isocratic
Temperature: 45 °C
Volume: 5 µl, Chromatogram for a mixture of anions with concentration of 1 ppm
Substances: Chloride (Cl-), Nitrite (NO2-), Nitrate (NO3-)
Keywords: Anions
Chromatogram:
1. Chloride 1 ppm 1
2. Nitrite 1 ppm
3. Nitrate 1 ppm 2
3
0 5 10 15 20 min
V 09/2008 49
23 Determination of Organic Acids
Method Matrix
HPLC
Column: ProntoSIL 120-3 C18 AQ, 300 x 3 mm ID Order No. 30CF184PSG
Conditions: Eluent: 5 mM Li2SO4 / H2SO4 (pH 2.81)

Gradient: isocratic
Temperature: 20 °C
Volume: 10 µl
Substances: Acetic acid, Ascorbic acid, Citric acid, Fumaric acid, Lactic acid, Malic acid, Malonic
acid, Oxalic acid, Quinic acid, Shikimic acid, Succinic acid, Tartaric acid
Keywords: Organic Acids
Chromatogram:
1. Oxalic acid 1
2. not identified 3
3. Tartaric acid
8
4. Quinic acid
5. Malic acid
9
6. Malonic acid 2 4
7. Shikimic acid 7
10
8. Lactic acid
9. Ascorbic acid 5 6 12 13
10. Acetic acid
11
11. Citric acid
12. Fumaric acid
13. Succinic acid
0 5 10 min
24 Determination of Organic Acids II
Method Matrix
HPLC
Column: Eurokat H, 300 x 8 mm ID Order No. 30GX340EKN
Phase: Eurokat H (polymer phase)
Conditions: Eluent: 0.01 N Sulfuric Acid

Gradient: isocratic
Temperature: 75 °C
Volume: 20 µl
Substances: Malic Acid, Lactic Acid, Quinic Acid, Shikimic Acid
Keywords: Organic Acids
Chromatogram: 3
1. 1 Malic acid
2. Quinic acid 2
3. Shikimic acid 1
4. Lactic acid
0 10 20 30 min
V 09/2008 51
25 Determination of Patulin in apple juice
Method Matrix
HPLC Apple juice
Sample extraction of apple juice with ethyl acetate (3x), extraction of the combined phases with
preparation: Na2CO3 solution, extraction of the separated aqueous phase with ethyl acetate,
addition of glacial acetic acid, reduction of ethyl acetate, redissolution of the dry
residue in mobile phase

Gradient: Isocratic
Temperature: 25 °C
Volume: 10 µl
Detection: UV (DAD) at 275 nm
Substances: Patulin
Keywords: Mycotoxins
Chromatogram:
H
H
O
H
O
O
H H
OH
Patolin
Patulin
0 5 10 15 min
26 Separation of Preservatives
Method Matrix
HPLC
Conditions: Eluent: A: Buffer / Methanol (50:20) B: Buffer / Methanol (50:70) Buffer:

0.4 ml Formic Acid, 0.8 ml NH3 (25 %) filled with Water to 1l
Gradient: 0 – 15 min: 100 % A to 100 % B
15 – 20 min: 100 % B
Temperature: 30 °C
Volume: 20 µl
Detection: UV at 235 nm and 255 nm
Substances: Benzoic acicd, 4-Hydroxy-benzoic acid, p-Hydroxy methylbenzoate, p-Hydroxy

ethylbenzoate, p-Hydroxy propylbenzoate, p-Hydroxy butylbenzoate, 2-Methoxy
benzoic acid, Sorbic acid
Keywords:
Chromatogram: 4
1. 4-Hydroxy benzoic acid
2. 2-Methoxy benoic acid
3. Benzoic acid
4. Sorbic acid
5. p-Hydroxy methylbenzoate
6. p-Hydroxy ethylbenzoate
7. p-Hydroxy propylbenzoate 1
8. p-Hydroxy butylbenzoate 5
6 7
8
3
2
255 nm / 235 nm
0 5 10 15 20 min
V 09/2008 53
27 Determination of various Sterols
Method Matrix
HPLC
Conditions: Eluent: A: Water B: Acetonitrile / Methanol (99:1)

Gradient: isocratic 10 % A and 90 % B
Temperature: 25 °C
Volume: 2 µl
Substances: Ergosterol, Lanosterol, Cholesterol, Campesterol, Stigmasterol, Sitosterol
Keywords: Steroids, Steroles
Chromatogram: 5
1. Ergosterol 4
2. Lanosterol
3. Cholesterol
4. Campesterol 3
5. Stigmasterol
6. Sitosterol
1
2
0 5 10 15 min
28 Separation of water soluble Vitamins
Method Matrix
HPLC
Column: ProntoSIL 120-3-C18 AQ, 250 x 3 mm Order No. 25CF184PSG
Phase: ProntoSIL 120-3-C18 AQ
Conditions: Eluent: A: 50 mM H3PO4 in Water B: Acetonitrile

Gradient: 0 – 3 min 100 % A
3 – 6.66 min 100 – 70 % A
6.66 - min 70 % A
Temperature: 25 °C
Volume: 5 µl
Substances: Pyridoxamine, Thiamine (Vit. B1), Ascorbic acid (Vit. C), Nicotinic acid (Niacin)
Nicotinamide (Vit. B3), Pyridoxale, Pyridoxine (Vit. B6), Folic acid, Cyanocobalamine
(Vit. B12), Riboflavine (Vit. B2)
Keywords: Vitamins, water soluble Vitamins
Chromatogram:
9
1. Pyridoxamine
2. Thiamine (Vit. B1)
3. Ascorbic acid (Vit. C)
4. Nicotinic acid
5. Nicotinamide (Vit. B3)
6. Pyridoxale
7. Pyridoxine (Vit. B6) 10
8. Folic acid,
9. Cyanocobalamine (Vit. B12) 4 6
10. Riboflavine (Vit. B2) 1
2 3 8
7
5
0 5 10 min
V 09/2008 55
29 Separation of Tocopherols and Vitamin D2
Method Matrix
HPLC
Column: Eurospher 100-5 Si, 150 x 4 mm ID Order No. 15DE000ESJ
Phase: Eurospher 100-5 Si
Conditions: Eluent: Hexan / 2-Butanol (1000:4)

Gradient: isocratic
Temperature: 25 °C
Volume: 20 µl
Substances: Tocopherols (alpha, beta, gamma, delta), Vitamin D2
Chromatogram:
1
1. alpha-Tocopherol
2. beta-Tocopherol 2
3. gamma-tocopherol 3
4. delta-Tocopherol 4
5. Vitamin D2 5
6. trans-Retinol
0 5 10 15 20 25 min
30 Separation of fat soluble Vitamins
Method Matrix
HPLC
Column: ProntoSIL 120-3-C18 SH, 250 x 3 mm ID Order No. 25CF180PSG
Phase: ProntoSIL 120-3-C18 SH
Conditions: Eluent: Methanol

Gradient: isocratic
Volume: 5 µl (50 – 300 ppm each substance)
Detection: Evaporated Light Scattering Detector (T: 33 °C) / UV at 280 nm
Substances: Vitamin A acetate, Vitamin D3, Vitamin K1, Vitamin A alcohol, Vitamin D2, Vitamin E
Chromatogram:
5
1. Vitamin A acetate
2. Vitamin D3
3. Vitamin K1
4. Vitamin A alcohol
5. Vitamin D2
6. Vitamin E
1 2
34
0 5 10 15 20 min
V 09/2008 57
31 Fat soluble Vitamins by HPLC with electrochemical
detection
Method Matrix
HPLC carrot juice
Sample vitamins in carrot juice, extracted with methanol

Preparation:
Conditions: Eluent: Methanol with 2 g/l LiClO4 and 1 g/l Acetic Acid
Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl (20 ng of each standard)
Detection: electrochemical, glassy carbon working electrode,

left +1000 mV, right +800 mV
Substances: Vitamin A Acetate, α-Tocopherol, β-Carotene
Keywords: Vitamins
Chromatogram: 1
1. Vitamin A Acetate
2. α-Tocopherol
3. β-Carotene
0 5 10 15 min
58
Pharmaceutical Applications
Page
Quality control mixture of Acetyl Cystein 60
Separation and determination of Opium Alkaloids 61
HPLC method for the determination of Amineptine and its main Metabolite in
human plasma 62
Determination of Amino Acids with OPA 63
Separation of Aniline derivatives with electrochemical detection 64
Separation of Anilines and Sulfonamides by HPLC with electrochemical
detection 65
Separation of Trimethoprim and Sulfamethoxazol 66
Determination of Anticonvulsant drugs and Methyl Xanthine derivatives in
serum 67
Separation of Antiepileptics in serum 68
Separation of Antirheumatic drugs 69
Separation of Catecholamines 70
Determination of Catecholamines II 71
Clinical analysis of Chloramphenicol 72
Determination of Citrate and Oxalate in human urine 73
Determination of Guaifenesin, Codein and Pseudoefedrin in cough syrup 74
Isocratic separation of the Digoxin and Digitoxin series of Glycosides 75
Separation of Narcotics 76
Analysis of β-Lactam Antibiotic (Moxalactam) 77
Analysis of Bisphosphonate Pamidronate Disodium by precolumn derivatization
and fluorescence detection 78
Determination of Paracetamol and by-products 79
Separation of Penicillin Antibiotics 80
Separation of Peptides 81
Fast separation of Proteins 82
Separation of Sedative 83
Separation of Steroids 84
Separation of Steroids II 85
Separation of Sulfa drugs 86
Separation of Sulfa drugs II 87
Separation of Tricyclic Antidepressant drugs 88
Separation of D,L-Tryptophan and Aromatic Acids 89
59
1 Quality control mixture of Acetyl Cystein
Method Matrix
HPLC
Conditions: Eluent: 1.6 g Heptansulfonic Acid in 960 ml H2O (pH with H3PO4 85%)
and 40 ml MeOH
Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Detection: UV at 240 nm (Ascorbic acid); UV at 216 nm
Substances: Ascorbic acid, Citric acid, Adipic acid, Acetyl cystein (ACC), Cystein
Keywords: Organic Acids, Cystein, ACC
Chromatogram: 1
1. Ascorbic acid 2 4
2. Citric acid 3 5
3. Adipic acid
4. Acetyl cystein
5. Cystein
0 10 20 30 min
60 Pharmaceutical Applications
2 Separation and determination of Opium Alkaloids
Method Matrix
HPLC opium
Column: Eurospher 100-5 C18, 250 x 4mm ID Order No. 25DE181ESJ
Conditions: Eluent: 1 % Ammonium Acetate pH 5.8 / Acetonitrile (65:35)

Gradient: isocratic
Temperature: 25 °C
Volume: 20 µl
Substances: Morphine, Codeine, Cryptopine, Thebaine, Papaverine, Narcotine
Keywords: Alkaloids
Chromatogram: 6
2
1. Morphine
2. Codeine
3. Cryptopine 1 5
4. Thebaine
5. Papaverine
6. Narcotine 3
0 5 10 15 min
V 09/2008 61
3 HPLC method for the determination of Amineptine and its
main Metabolite in human plasma
Method Matrix
HPLC plasma
Conditions: Eluent: Aqueous phase containing 1.2 g/l heptane sulphonate adjusted to
pH 3.0 with H3PO4 / Acetonitrile (62:38)
Gradient: isocratic
Temperature: 30 °C
Volume: 20 µl
Substances: Amineptine
Keywords: Drugs, Antidepressant drugs
Chromatogram:
Chromatograms of extracts from
a) blank plasma a b c
b) plasma spiked with:

1. 250 ng/ml metabolite
2. 250 ng/ml amineptine
3. 500 ng/ml internal standard 1 1
c) a patient plasma sample
obtained 45 min after taking a
100 mg oral dose of 2 3 3
amineptine, spiked with 2
1. 230 ng/ml metabolite
2. 180 ng/ml amineptine
3. 500 ng/ml internal standard
0 5 10 0 5 10 0 5 10 min
4 Determination of Amino Acids with OPA
Method Matrix
HPLC
Column: ProntoSIL 120-3-C18 H, 250 x 3.0 mm ID Order No. 25CF185PSG
Phase: ProntoSIL 120-3-C18 H
Sample Amino Acid standard with OPA precolumn derivatization

Preparation:
Conditions: Eluent: A: 20 mM CH3COONa in H2O / ACN, (97:3)

B: 20 mM CH3COONa in H2O / ACN, (50:50)
Gradient: 5 – 28 % B, 0 – 144 s
28 – 45 % B, 145 – 560 s
45 – 82 % B, 561 – 896 s
82 – 90 % B, 897 – 1215 s
Temperature: 30 °C
Volume: 1 µl
Detection: Fluorescence: excitation 330 nm emission 450 nm

UV at 330 nm
Substances: Aspartic acid, Glutamic acid, Serine, Glutamin, Histidine, Glycine, Threonine, Arginine,
Alanine, Tyrosine, Methionine, Valine, Phenylalanine, Isoleucine, Leucine, Lysine
Keywords: Amino Acids
Chromatogram: 7
4
1. Asp
2. Glu 5 89 15
3. Ser 14
4. Gly 10 12
11 13
5. Thr
6. His 2
7. Ala 3
8. Arg 1
9. Tyr
10. Val
11. Cys 6
12. Met 16
13. Ile
14. Phe
15. Leu
16. Lys
0 5 10 15 20 min
V 09/2008 63
5 Separation of Aniline derivatives with electrochemical
detection
Method Matrix
HPLC
Conditions: Eluent: Methanol / Water (1:1) with 2 g/l KNO3 and 0.05 g/l H2SO4
Gradient: isocratic
Volume: 20 µl
Detection: Amperometric detection, glassy carbon 1000 mV / 800 mV, 100 nA
Substances: Phenylene diamine, Chloroaniline
Keywords: Amines
Chromatogram: 1
1. p-Phenylene diamine (20 ng)
2. 3-Chloroaniline (25 ng)
0 4 8 min
6 Separation of Anilines and Sulfonamides by HPLC with
electrochemical detection
Method Matrix
HPLC
Conditions: Eluent: Methanol / Water (1:1) with 2 g/l KNO3 and 0.05 g/l Sulfuric Acid
Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Detection: Amperometric detection, glassy carbon electrode, range 100 nA

+1000 mV for o-Toluidine / 1200 mV for sulfonamides,
Substances: Toluidine-o, Sulfanilamide, Sulfamethoxydiazine
Keywords: Drugs, Antibiotics
Chromatogram: 2
1. o-Toluidine
2. Sulfanilamide
3. Sulfmethoxydiazone
Upot = 1000 mV Upot = 1200 mV

3
0 4 8 min 0 4 8 min
V 09/2008 65
7 Separation of Trimethoprim and Sulfamethoxazol
Method Matrix
HPLC
Column: ProntoSIL 120-5 C18 SH, 250 x 4 mm ID Order No. 25DF180PSJ
Conditions: Eluent: Methanol / Water with 1 % formic acid (15:85)

Gradient: isocratic
Temperature: 30 °C
Volume: 10 µl
Substances: Trimethoprim, Sulfamethoxazol
Keywords: Drugs, Antibiotic
Chromatogram: 2
1 Trimethoprim
2. Sulfamethoxazol
0 5 10 15 20 min
8 Determination of Anticonvulsant drugs and
Methyl Xanthine derivatives in serum
Method Matrix
HPLC serum
Sample Precolumn loading and washing with Eluent A
preparation:
Conditions: Eluent: A: 14 mM NaH2PO4 - 6mM Na2HPO4

B: Eluent A / Acetonitrile / Methanol (6.5:1.5:2)
Gradient: isocratic
Temperature: 25 °C
Volume: 20 µl
Substances: Primidone, Phenobarbital, Phenytoin, Carbamazepine
Keywords: Drugs, Anticonvulsant drugs
Chromatogram:
1. Primidone (10 µg/ml)
2. Phenobarbital (10 µg/ml
3. Phenytoin (20 µg/ml)
4. Carbamazepine (5 µg/ml)
2 3 4
1
0 10 20 30 min
V 09/2008 67
9 Separation of Antiepileptics in serum
Method Matrix
HPLC
Column: Eurospher 100-5-C18, 100 x 4.0 mm ID Order No. 10DE181ESJ
Phase: Eurospher 100-5-C18
Conditions: Eluent: Recipe eluent: Antiepileptics in serum (Order no. 15010)

Gradient: isocratic
Temperature: 55 °C
Volume: 10 µl
Substances: PEMA, Ethosuximide, Primidone, Carbamazepine-diol, 10-OH-Carbamazepine,

Phenobarbital, Carbamazepine-epoxide, Oxcarbazepine, DPH, Carbamazepine
Keywords: Antiepileptics, Drugs
Chromatogram:
1. PEMA
2. Ethosuximide
3. Primidone
4. Carbamazepine-diol
5. 10-OH-Carbamazepine
6. Phenobarbital
7. Carbamazepine-epoxide
8. Oxcarbazepine 5
9. Internal Standard (IS) 2
10. DPH 9
11. Carbamazepine 1 6
3
8
4 7 10 11
0 10 20 30 40 min
10 Separation of Antirheumatic drugs
Method Matrix
HPLC
Conditions: Eluent: Methanol / Water (70:30), 0.1 % Acetic Acid with

6 g/l Ammonium Acetate and 1.5 g/l Sulfuric Acid
Gradient: isocratic
Temperature: 20 °C
Volume: 20 µl
Substances: Camphor, Ethylenglycolmonosalicylate, Nicotinic acid methyl ester, Nicotinic acid

benzyl ester
Keywords: Antirheumatic drugs
Chromatogram: 3
1. Camphor
2. Nicotinic acid methyl ester
3. Nicotinic acid benzyl ester
4. Ethylenglycolmonosalicylate
0 5 10 min
V 09/2008 69
11 Separation of Catecholamines
Method Matrix
HPLC
Column: UltraSep ES CAQC, 125 x 3 mm ID Order No.I0203
Phase: UltraSep ES CAQC
Conditions: Eluent: prepared standard eluents: CA

Gradient: isocratic
Flow rate: 1 ml/min
Temperature: 25 °C
Volume: 5 µl
Substances: Vanillyl mandelic acid (VMA); Iso-VMA; 3,4-Dihydroxyphenylic acetic acid (DOPAC); 5-
Hydroxyindolyl acetic acid (5-HIAA); Homovaniliic acid (HVA); 5-HICA; Dopamine; 3-
Methoxytyramine (3-MT); 5-Hydroxytryptamine (Serotonine)
Keywords: Catecholamines
Chromatogram:
1. Vanillyl mandelic acid (VMA)
2. Iso-VMA
3. 3,4-Dihydroxyphenylic acetic acid (DOPAC)
4. 5-Hydroxyindolyl acetic acid (5-HIAA)
5. Homovanillic acid (HVA)
6. 5-Hydroxyindol-2-carboxylic acid (5-HICA)
7. 3-Hydroxytyramine (Dopamine)
8. 3-Methoxytyramine (3-MT)
9. 5-Hydroxytryptamine (Serotonine)
12 Determination of Catecholamines II
Method Matrix
HPLC
Conditions: Eluent: prepared standard eluent “Urinary catechoamines”

Gradient: isocratic
Temperature: 60 °C
Detection: Amperometric, Cell potential 0.55 V, 10 nA/V, glassy carbon electrode
Substances: Adrenaline, Noradrenaline, Dihydrjoxybenzylamine (IS); Dopamine
Keywords: Catecholamines
Chromatogram:
1. Noradrenaline
2. Adrenalina
3. Dihydroxybenzylamine (IS)
4. Dopamine
1 4
0 10 20 30
V 09/2008 71
13 Clinical analysis of Chloramphenicol
Method Matrix
HPLC serum
Column: Eurospher 100-5 C18, 100 x 2mm ID Order No. 10BE181ESJ
Conditions: Eluent: 0.1 N Acetate buffer (pH 6.0) / Acetonitrile (8:2)

Gradient: isocratic
Volume: 0.5 µl
Substances: Chloramphenicol
Keywords: Drugs, Antibacterial drugs
Chromatogram:
(a) 10 mg/l Chloramphenicol
(b) 25mg/l Chloramphenicol

2
1. Chloramphenicol
2. Internal Standard
2
(a) (b)
0 5 10 0 5 10 15 min
14 Determination of Citrate and Oxalate in human urine
Method Matrix
HPLC urine
Column: Novosep A-2 Anion, 5µm, 250 x 4 mm ID Order No. B92
Conditions: Eluent: 15 mM Sodium Carbonate (Na2CO3)

Gradient: isocratic
Temperature: 50 °C
Volume: 5 µl, urine sample 1:20 (blue)
Substances: Citrate, Oxalate
Keywords: Organic Anions
Chromatogram: 1
1. Chloride
2. Phosphate
3. Sulfate
4. Oxalate
5. Citrate
4
5
0 5 10 15 20 min
V 09/2008 73
15 Determination of Guaifenesin, Codein and Pseudoefedrin
in cough syrup
Method Matrix
HPLC cough syrup
Column: ProntoSIL 120-3 C18 SH, 250 x 4 mm ID Order No. 25EK185PSJ

25EK185PSJ
Conditions: Eluent: Water / Methanol (70:30) with 0.1 % TFA (pH 1.87)
Gradient: isocratic
Temperature: 40 °C
Volume: 5 µl
Substances: Codein, Guaifenesin, Pseudoefedrin
Keywords: Codein, Guaifenesin, Pseudoefedrin
Chromatogram: 3
1. Codein
2. Pseudoefedrin
3. Guaifenesin
2
1
n.i.
0 5 10 15 20 min
16 Isocratic separation of the Digoxin and Digitoxin series of
Glycosides
Method Matrix
HPLC
Column: Eurospher 100-5 C18, 250 x 4mm ID Order No. 25DE181ESJ
Conditions: Eluent: Acetonitrile / Methanol / Water (30:30:40)

Gradient: isocratic
Temperature: 30 °C
Volume: 10 µl
Substances: Digoxigenin, Digoxigenin monodigitoxoside, Digoxigenin bisdigitoxoside, Digoxin,

Digitoxigenin, Digitoxigenin monodigitoxoside, Digitoxigenin bisdigitoxoside, Digitoxin
Keywords: Drugs, Glycosides, Digitalis Glycosides
Chromatogram:
1
1. Digoxigenin
2. Digoxigenin monodigitoxoside
3. Digoxigenin bisdigitoxoside 2
4. Digoxin
5. Digitoxigenin
6. Digitoxigenin monodigitoxoside 5
7. Digitoxigenin bisdigitoxoside
8. Digitoxin
3
4 6
0 5 10 15 20 min
V 09/2008 75
17 Separation of Narcotics
Method Matrix
HPLC
Column: Eurospher 100-3 C18, 150 x 4 mm ID Order No. 15DE181ESG
Conditions: Eluent: Water / ACN (45:55)

Gradient: isocratic
Flow rate: 1.0 ml/min,
Temperature: 40 °C
Volume: 2 µl
Substances: Veronal, Luminal, Prominal, Revonal, Thiogenal
Keywords: Drugs, Narcotics
Chromatogram: 4
1. Barbital
2. Luminal
3. Prominal
4. Revinal
1
2 3
0 5 10 min
18 Analysis of β-Lactam Antibiotic (Moxalactam)
Method Matrix
HPLC
Conditions: Eluent: Methanol / 0.05 M Kalium phosphate (5:95) adjusted to pH 6.5

Gradient: isocratic
Temperature: 20 °C
Volume: 10 µl
Substances: Moxalactam, Methyl-1H-tetrazole-5-thiol
Keywords: Drugs, Antibiotics
Chromatogram: 2
1. 1-Methyl-1H-tetrazole-5-thiol
2. R-Moxalactam
3. S-Moxalactam
3
0 10 20 30 min
V 09/2008 77
19 Analysis of Bisphosphonate Pamidronate Disodium
by precolumn derivatization and fluorescence detection
Method Matrix
HPLC urine
Conditions: Eluent: A) 1 mM aqueous Na2EDTA, adjusted to pH 6.5 with 1 M NaOH B)

Methanol (97:3)
Gradient: isocratic
Volume: 20 µl

a. drug-free human urine b. drug-free dog urine
c. urine from a dog receiving a single intravenous injection of 5 mg of pamidronate
disodium in 0.15 M saline
Substances: Pamidronate disodium,

Disodium 3-amino-1-hydroxypropylidenebisphosphonate pentahydrate (IS)
Keywords: Drugs
Chromatogram: 3
1. Injection Peak
2. Pamidronate
3. Internal Standard (IS)
2
1
a) b) 1 c) 1
0 5 10 15 min 0 5 10 15 min 0 5 10 15 min
20 Determination of Paracetamol and by-products
Method Matrix
HPLC
Column: ProntoSIL 120-5 C8 ace-EPS, 250 x 3 mm ID Order No. 25CF08APSJ
Phase: ProntoSIL 120-5 C8 ace-EPS
Conditions: Eluent: A: ACN B: H2O (pH 2.75 with H3PO4)

Gradient: 0 – 0.65 min 10 % A
0.65 – 6.4 min 10 Æ 60 % A
6.4 – 10.4 min 60 % A
10.4 - 12 min 10 % A
Temperature: 50 °C
Volume: 5µl
Substances: Paracetamol, N-(2-hydroxyphenyl)acetamide, N-(4-hydroxyphenyl)propanamide,

N-phenylacetamide, Chloracetanilide, 1-(4-hydroxyphenyl)ethanone oxime,
1-(2-hydroxyphenyl)ethanone, 4-Aminophenol, 4-Nitrophenol
Keywords: Paracetamol
Chromatogram: 2
1. 4-Aminophenol
2. Paracetamol 4
3. N-(2-hydroxyphenyl)acetamide
4. N-(4-hydroxyphenyl)propanamide
5. N-phenylacetamide
8
6. 1-(4-hydroxyphenyl)ethanone oxime
7. 1-(2-hydroxyphenyl)ethanone 1
8. Chloracetanilide 5
6 7
9. 4-Nitrophenol
3
9
0 2.5 5 7.5 min
V 09/2008 79
21 Separation of Penicillin Antibiotics
Method Matrix
HPLC

25DE181ESJ
Conditions: Eluent: ACN / H2O (0.1 % TFA) (50:50)

Gradient: isocratic
Temperature: 25 °C
Volume: 1 µl
Substances: Ampicillin, Penicillin G, Penicillin V, Nafcillin, Dicloxacillin
Keywords: Drugs, Antibiotics, Penicillines
Chromatogram: 1
1. Ampicillin 4
2. Penicillin G
3. Penicillin V 3
4. Nafcillin
5. Dicloxacillin
2
5
0 5 10 min
22 Separation of Peptides
Method Matrix
HPLC
Column: ProntoSIL 300-5 C18 SH, 250 x 4.6 mm ID Order No. 25EK185PSJ
Conditions: Eluent: A: H2O / 0.1 % TFA B: ACN / H2O (70:30) + 0.1 % TFA
Gradient: 20-100 % B in 30 min
Temperature: 30 °C
Volume: 5 µl
Substances: Angiotensin I und II, Bradykinin, Eledoisin, Neurotensin, Oxytocin
Keywords: Peptides
Chromatogram: 2
1. Oxytocin
2. Bradykinin
3. Angiotensin II
4. Eledoisin
5. Neurotensin 4
6. Angiotensin I
1 3
0 5 10 15 20 min
V 09/2008 81
23 Fast separation of Proteins
Method Matrix
HPLC
Column: ProntoSIL 300-3 C18 SH, 75 x 4.6 mm ID Order No. 0746K185PS030

(*B)
Conditions: Eluent: A: H2O / 0.1 % TFA B: ACN / H2O (95:5) + 0.1 % TFA
Gradient: 0-25 s 29 - 48% B
25-65 s 48 - 100% B
Temperature: 30 °C
Volume: 5 µl
Substances: BSA, Insulin bovine, Lysozyme, Myoglobin, Ovalbumin, Ribonuclease A
Keywords: Proteins
Chromatogram: 3
1. Ribonuclease A
2. Insulin bovine 5
3. Lysozyme
4. BSA
5. Myoglobin
6. Ovalbumin 6
7. Not identified
4
1
2 7
0 1 2 min
24 Separation of Sedative
Method Matrix
HPLC
Column: Eurospher 100-5 C18, 250 x 4.6 mm Order No. 25EE181ESJ
Conditions: Eluent: A: Methanol B: Water (60:40)

Gradient: isocratic
Volume: 10 µl
Substances: Barbital, Luminal, Prominal, Revonal, Thiogenal
Keywords: Sedativa
Chromatogram: 1
1. Barbital 2
2. Luminal
3. Prominal 3
4. Revonal
5. Thiogenal
4
0 5 10 15 20 min
V 09/2008 83
25 Separation of Steroids
Method Matrix
HPLC
Conditions: Eluent: A: Methanol B: Water

Gradient: 0 - 35 min 50 % Æ 95 % A
35 - 45 min 95 % Æ 50 % A
Temperature: 30 °C
Volume: 10 µl
Substances: Cortisone, Corticosterone, Desoxycorticosterone, Testosterone, Norgestrel,

Progesterone,
Keywords: Steroids
Chromatogram: 4
1. Cortisone 2
2. Corticosterone
3. Desoxycorticosterone
4. Testosterone 3
1 6
5. Norgestrel 5
6. Progesterone
0 10 20 30 min
26 Separation of Steroids II
Method Matrix
HPLC
Column: ProntoSIL 120A-5 C8 SH, 125 x 4 mm ID Order No. 12DF08PSJ
Phase: ProntoSIL 120A-5 C8 SH
Conditions: Eluent: A: H2O B: MeOH

Gradient: 80 % A (1 min hold), 65 % A in 10 min, 65 % A (3 min),
40 % A in 6 min, 40 % A 4 min hold
Temperature: 30 °C
Volume: 10 µl
Substances: Esteriol, Prednisolone, Cortisone, Testosterone, Methyl-6α-hydroxy-11β-progesterone,

Methyl-6α-hydroxy-17α-progesterone, Methyl-6α-hydroxy-17α-progesterone acetate,
Estradiol, Estrone, Progesterone
Keywords: Steroids
Chromatogram: 6
1. Esteriol
2. Prednisolone
3. Cortisone
4. Testosterone
5. Methyl-6α-hydroxy-11β-progesterone
6. Estradiol
7. Methyl-6α-hydroxy-17α-progesterone 10
8. Methyl-6α-hydroxy-17α-progesterone acetate 3
9. Estrone 4 9
10. Progesterone
2 7
8
5
1
0 5 10 15 20 min
V 09/2008 85
27 Separation of Sulfa drugs
Method Matrix
HPLC
Column: ProntoSIL 120-5 C18 ace-EPS, 150 x 4.6 mm ID Order No. 15CE181ESG
Phase: ProntoSIL 120-5 C18 ace-EPS
Conditions: Eluent: Methanol / Water / Acetic Acid (20:79:1) with

6 g/l Ammonium Acetate and 1.5 g/l Sulfuric Acid
Gradient: isocratic
Temperature: 25 °C
Volume: 3 µl
Substances: Sulfanilamide, Sulfadiazine, Sulfathiazole, Sulfamerazine, Sulfamethazine
Keywords: Sulfa drugs
Chromatogram: 5
4
1. Sulfanilamide
2. Sulfadiazine
3. Sulfathiazole 1
4. Sulfamerazine
5. Sulfamethazine
0 5 10 min
28 Separation of Sulfa drugs II
Method Matrix
HPLC
Column: Eurospher 100-3 C18, 150 x 3 mm ID Order No. 15CE181ESG
Conditions: Eluent: A: ACN

B: 0.2 g NaH2PO4 in 1000 ml H2O (pH 4 with H3PO4)
Gradient: 22 % A 0 – 3.5 min
30 % A 3.5 – 4.5 min
90 % A 4.5 – 6 min, 2 min hold
Flow rate: 0 - 3.5 min 0.2 ml/min, 3.5 - 10 min 0.4 ml/min
Temperature: 40 °C
Volume: 10 µl
Substances: Sulfadiazine, Sulfamethiazole, Sulfamethoxypyridazine, Sulfamethoxazole,

Sulfadimethoxine
Keywords: Sulfa drugs, antibiotics
Chromatogram: 4
1. Sulfadiazine
2. Sulfamethiazole
3. Sulfamethoxypyridazine 2 3 5
4. Sulfamethoxazole
5. Sulfadimethoxine
1 5 10 m in
V 09/2008 87
29 Separation of Tricyclic Antidepressant drugs
Method Matrix
HPLC
Column: ProntoSIL 120-3-C18 AQ, 60 x 4.0 mm ID Order No. 06DF184PSG
Phase: ProntoSIL 120-3-C18 AQ
Conditions: Eluent: ACN / MeOH / 10 mM Phosphate Buffer (62:13:25)

Gradient: isocratic
Temperature: 20 °C
Volume: 5 µl
Substances: Amitriptyline, Doxepin, Imipramine, Nortiptyline, Trimipramine
Keywords: Antidepressant, Drugs
Chromatogram: 2
1. Doxepin
2. Imipramin
3. Nortriptyline
4. Amitriptyline
5. Trimipramine
3
5
4
0 5 10 15 min
30 Separation of D,L-Tryptophan and Aromatic Acids
Method Matrix
HPLC
Conditions: Eluent: Methanol / Water (30:70)

with 6 g/l Ammonium Acetate and 1.5 g/l Sulphuric Acid
Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl (40 ng each)
Detection: Amperometric detection, +1000 mV / 800 mV, range 100 nA
Substances: Dihydroxyphenylacetic acid, Tryptophan, Homovanillic acid
Keywords: Biogenic amines, aromatic acids
Chromatogram: 1
1. 3,4-Dihydroxyphenylacetic acid,
2. D,L-tryptophan,
3. Homovanillic acid 3
0 5 10 15 20 min
V 09/2008 89
Chiral Applications
(HPLC)
Page
Chiral separation of Abscisic Acid (4-Oxo-2-Cyclohexen-1-yl)-3-Methyl-
2,4-Pentanedienoic Acid) 92
Chiral separation of 3-Amino-2(2-Naphtyl)-1-Phenyl-Propanol 93
Chiral separation of Atenolol (2-[4-[2-Hydroxy-3-
(1-Methylethylamino)Propoxy]Phenyl]Ethanamid) 94
Chiral separation of Atropine (rac-Hyoscyamin) 95
Chiral separation of 1-Aza[6]Helicene 96
Chiral separation of 1-Benzene-2-Naphthene-3-Amino Propan(1)ol 97
Chiral separation of Benzoin (α-Hydroxy-α-Phenylacetophenon) 98
Chiral separation of Benzyl-Mandelate (Benzyl-2-Hydroxy-3-Phenylpropionate) 99
Chiral separation of 2,2´Bis(Diphenylphosphinoxid)3,3´Bibenzo[b]Thiophene 101
Chiral separation of BITIANP
2,2´Bis(Diphenylphosphino)3,3´Bibenzo[b]Thiophene 102
Chiral separation of 4-Bromphenyl-Glycinamid (2-(4-Bromphenyl)-2-Amino
Acetamid) 103
Chiral separation of Carbinoxamine (2-[(4-Chlorophenyl)-Pyridin-2-yl-Methoxy]-
N,N-Dimethyl-Ethanamine) 104
Chiral separation of Citalopram (1-(3-Dimethylaminopropyl)-1-(4-Fluorphenyl)-
3H-2-Benzofuran-5-Carbonitrile) 105
Chiral separation of Dilactid 106
Chiral separation of 1,2-Diphenyl-2-(Tosylamino)Ethanone 107
Chiral separation of Ethyl-Mandelate 108
Chiral separation of Etozolin (Ethyl(3-Methyl-4-Oxo-5-Piperidinothiazolidin-
2-ylidene)Acetate) 109
Chiral separation of Flavanone (2-Phenyl-1,4-Benzopyrone) 110
Chiral separation of Hexahydro-6,6-Dimethyl-2-Tosylpyrrolo
[1,2-e]Imidazol-3-one 111
Chiral separation of Hydroxyzine 112
Chiral separation of Lactic Acid 113
Chiral separation of n-Methylephedrin (2-Dimethylamino-1-Phenyl-Propanol) 114
Chiral separation of Methyl phenyl sulfoxide 115
Chiral separation of Methyl phenyl sulfoxide II 116
90
Chiral separation of Metoprolol (1-(Isoppropylamino)-3-[4-(2-
Methoxyethyl)Phenoxy]Propan-2-ol) 117
Chiral separation of 1-(Naphthalene-6-yl)Ethane-1,2-diol 118
Chiral separation of Naproxen (6-Methoxy-α-Methyl-2-Naphthylessigsäure) 119
Chiral separation of 1-Phenylethylenglycol (1-Phenylethane-1,2-diol) 120
Chiral separation of 1-Phenylpropane-1,2-diol 121
Chiral separation of Pindolol (1-(1H-Indol-4-yloxy)-
3-(1-Methylethylamino)Propan-2-ol) 122
Chiral separation of Pindolol (1-(1H-Indol-4-yloxy)-
3-(1-Methylethylamino)Propan-2-ol) 123
Chiral separation of Propranolol (1-Isopropylamino-3-(1-Naphthyloxy)-
2-Propanol) 124
Chiral separation of Propranolol (1-Isopropylamino-3-(1-Naphthyloxy)-
2-Propanol) 125
Chiral separation of Warfarin (4-Hydroxy-3-(3-Oxo-1-Phenyl-Butyl)-Cumarin) 126
V 09/2008 91
1 Chiral separation of Abscisic Acid (4-Oxo-2-Cyclohexen-
1-yl)-3-Methyl-2,4-Pentanedienoic Acid)
Method Matrix
HPLC Reversed Phase Mode
Column: Eurocel 01, 5 µm, 250 x 4.6 mm ID Order No. 25EM370ECJ
Phase: Eurocel 01
Conditions: Eluent: Methanol / Water (50:50) + 0.1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 5 µl
Substances: Abscisic acid (4-Oxo-2-Cyclohexen-1-yl)-3-Methyl-2,4-Pentanedienoic Acid)
Keywords: plant hormone, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
CH3 CH3 CH3
k´1 = 2.67
k´2 = 3.19 OH
α = 1.19
O
O CH3 OH
92 Chiral Applications (HPLC)

2 Chiral separation of 3-Amino-2(2-Naphtyl)-1-Phenyl-
Propanol
Method Matrix
HPLC Normal Phase Mode
Phase: Eurocel 01
Conditions: Eluent: Heptane / 2-Propanol (90:10) + 0.1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 3-Amino-2(2-Naphtyl)-1-Phenyl-Propanol
Keywords: chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
k´1 = 2.46
k´2 = 3.13 OH
α = 1.27
NH2
0 5 10 15 20 min
V 09/2008 93
3 Chiral separation of Atenolol (2-[4-[2-Hydroxy-3-
(1-Methylethylamino)Propoxy]Phenyl]Ethanamid)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Heptane / 2-Propanol (80:20) + 0.1 % EtOA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Atenolol (2-[4-[2-Hydroxy-3-(1-Methylethylamino)Propoxy]Phenyl]Ethanamid)
Chromatogram:
NH2
k´1 = 6.53 NH
k´2 = 8.17 O
α = 1.25 O
OH
0 10 20 30 min

4 Chiral separation of Atropine (rac-Hyoscyamin)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Hexane / 2-Propanol (80:20)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Atropine
Chromatogram: OH
N
k´1 = 0.54
k´2 = 0.89 O
α = 1.65
O
0 5 10 15 min
V 09/2008 95
5 Chiral separation of 1-Aza[6]Helicene
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Heptane / 2-Propanol (75:25)

Gradient: isocratic
Volume: 20 µl
Substances: 1-Aza[6]Helicene
Chromatogram:
k´1 = 1.28
k´2 = 1.59 N
α = 1.24
0 5 10 15 min

6 Chiral separation of 1-Benzene-2-Naphthene-3-Amino
Propan(1)ol
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Heptane / 2-Ppropanol (90:10) + 0.1 % TFA

Gradient: isocratic
Volume: 10 µl
Substances: 1-Benzene-2-Naphthene-3-Amino Propan(1)ol
Chromatogram:
k´1 = 2.46 OH
k´2 = 3.13
NH2
α = 1.27
0 10 20 30 min
V 09/2008 97
7 Chiral separation of Benzoin (α-Hydroxy-α-
Phenylacetophenon)
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Volume: 10 µl
Substances: Benzoin, (α-Hydroxy-α-Phenylacetophenon)
Chromatogram:
O
k´1 = 7.18
k´2 = 11.33
α = 1.58 OH
0 5 10 15 20 min

8 Chiral separation of Benzyl-Mandelate (Benzyl-2-Hydroxy-
3-Phenylpropionate)
Method Matrix
HPLC Polar organic mode
Phase: Eurocel 01
Conditions: Eluent: Methanol + 0,1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Benzyl-Mandelate (Benzyl-2-Hydroxy-3-Phenylpropionate)
Chromatogram: OH
k´1 = 0.31 O
k´2 = 0.40
O
α = 1.29
350
300
250
200
150
100
50
0 1 2 3 4 5 6 7 8 9 10
Minutes
V 09/2008 99
3-Phenylpropionate)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Hexane / 2-Propanol (70:30)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Chromatogram: OH
k´1 = 0.75 O
k´2 = 1.62
O
α = 2.16
450
400
350
300
250
200
150
100
50
0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0 8,5 9,0 9,5
Minutes

10 Chiral separation of
2,2´Bis(Diphenylphosphinoxid)3,3´Bibenzo[b]Thiophene
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Hexan / 2-Propanol (85:15)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 2,2´Bis(Diphenylphosphinoxid)3,3´Bibenzo[b]Thiophene
Chromatogram:
k´1 = 2.45
S
k´2 = 2.99
P
α = 1.22 O
O
S
600
500
400
mAU 300
200
100
0 2 4 6 8 10 12 14 16 18 20
Minutes
V 09/2008 101
11 Chiral separation of BITIANP
2,2´Bis(Diphenylphosphino)3,3´Bibenzo[b]Thiophene
Method Matrix
HPLC Polar Organic Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: BITIANP (2,2´Bis(Diphenylphosphino)3,3´Bibenzo[b]Thiophene)
Keywords: BITIANP, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
k´1 = 1.47
S
k´2 = 1.71
P
α = 1.16
S
60
50
40
mAU
30
20
10
0 2 4 6 8 10 12 14 16 18 20
Minutes

12 Chiral separation of 4-Bromphenyl-Glycinamid
(2-(4-Bromphenyl)-2-Amino Acetamid)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Heptane / 2-Propanol (90:10) + 0.1 % DEA

Gradient: isocratic
Volume: 10 µl
Substances: 4-Bromphenyl-Glycinamid (2-(4-Bromphenyl)-2-Amino Acetamid)
Chromatogram: O NH2
k´1 = 3.09
k´2 = 4.54
NH2
α = 1.47
Br
0 5 10 15 min
V 09/2008 103
13 Chiral separation of Carbinoxamine (2-[(4-Chlorophenyl)-
Pyridin-2-yl-Methoxy]-N,N-Dimethyl-Ethanamine)
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Carbinoxamine (2-[(4-Chlorophenyl)-Pyridin-2-yl-Methoxy]-N,N-Dimethyl-Ethanamine)
Keywords: antihistamine, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
N CH Cl
k´1 = 1.69
k´2 = 1.94
OCH2CH2N(CH3)2
α = 1.14
0 5 10 min

14 Chiral separation of Citalopram (1-(3-
Dimethylaminopropyl)-1-(4-Fluorphenyl)-3H-2-Benzofuran-
5-Carbonitrile)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Heptane / 2-Propanol (96:4) + 0.1 % DEA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Citalopram (1-(3-Dimethylaminopropyl)-1-(4-Fluorphenyl)-3H-2-Benzofuran-5-Carbonitrile)
Keywords: therapeutical drug, chiral separation, Eurocel 01, cellulose based chiral selector
F
Chromatogram:
N
k´1 = 2.67
k´2 = 3.27
O
α = 1.23
N
0 5 10 15 min
V 09/2008 105
15 Chiral separation of Dilactid
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Hexane / Ethanol (90:10)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Dilactid (D-, L-, meso)
Chromatogram: CH3
O O
k´1 = 2.01
k´2 = 2.22
α = 1.10 O O CH3
0 5 10 15 min

16 Chiral separation of 1,2-Diphenyl-2-(Tosylamino)Ethanone
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 1,2-Diphenyl-2-(Tosylamino)Ethanone
Chromatogram:
O
k´1 = 7.66 H
N
Tos
k´2 = 8.66
α = 1.13
0 10 20 30 40 min
V 09/2008 107
17 Chiral separation of Ethyl-Mandelate
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Hexane / 2-Propanol (90:10)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Ethyl-Mandelate
Chromatogram:
OH
k´1 = 1.10
k´2 = 2.32
COOEt
α = 2.11
800
700
600
500
400
300
200
100
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Minutes

18 Chiral separation of Etozolin (Ethyl(3-Methyl-4-Oxo-5-
Piperidinothiazolidin-2-ylidene)Acetate)
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Etozolin (Ethyl(3-Methyl-4-Oxo-5-Piperidinothiazolidin-2-ylidene)Acetate)
Keywords: diureticum, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
k´1 = 0.68
k´2 = 1.05
N
α = 1.54
S O
O
N O CH3
CH3
0 3 6 9
V 09/2008 109
19 Chiral separation of Flavanone (2-Phenyl-
1,4-Benzopyrone)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Heptane / 2-Propanol (90:10)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Flavanone (2-Phenyl-1,4-Benzopyrone)
Keywords: antioxidants, chiral separation, Eurocel 01, cellulose based chiral selector
O
Chromatogram:
k´1 = 1.41
k´2 = 1.83
O
α = 1.30
0 3 6 9 12 15 min

20 Chiral separation of Hexahydro-6,6-Dimethyl-2-
Tosylpyrrolo [1,2-e]Imidazol-3-one
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Hexan / 2-Propanol (75:25)

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Hexahydro-6,6-Dimethyl-2-Tosylpyrrolo[1,2-e]Imidazol-3-one
Chromatogram:
O
k´1 = 3.92
N
k´2 = 4.92 N Tos
α = 1.25
0 10 20 30 40 50 min
V 09/2008 111
21 Chiral separation of Hydroxyzine
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Hydroxyzine
Chromatogram: Cl O
N OH
k´1 = 1.88 N
k´2 = 2.15
α = 1.14
0 10 15 min

22 Chiral separation of Lactic Acid
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Heptane / 2-Butanole (95:5)+ 0.1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Lactic Acid
Chromatogram:
OH
k´1 = 2.99
k´2 = 3.47
α = 1.16 CH3 CH COOH
180
160
140
120
100
80
60
40
20
-20
0 2 4 6 8 10 12 14 16 18 20
Minutes
V 09/2008 113
23 Chiral separation of n-Methylephedrin (2-Dimethylamino-1-
Phenyl-Propanol)
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: N-Methylephedrin (2-Dimethylamino-1-Phenyl-Propanol)
Keywords: amphetamine, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
OH CH3
k´1 = 0.88
k´2 = 1.25 N
α = 1.42
CH3
CH3
0 5 10 15 min

24 Chiral separation of Methyl phenyl sulfoxide
Method Matrix
Column: Eurocel 01, 3 µm, 250 x 4.6 mm ID Order No. 25EM370ECG
Phase: Eurocel 01
Conditions: Eluent: Hexane / Ethanol (90:10)

Gradient: isocratic
Flow rate: 1 ml/min
Temperature: 25 °C
Volume: 10 µl
Substances: Methyl phenyl sulfoxide
Keywords: Chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
O
k´1 = 4.65
S CH3
k´2 = 5.52
α = 1.18
0 5 10 15 20 25 30 min
V 09/2008 115
25 Chiral separation of Methyl phenyl sulfoxide II
Method Matrix
Phase: Eurocel 02
Conditions: Eluent: Hexane / Ethanol (75:25)

Gradient: isocratic
Flow rate: 1 ml/min
Temperature: 25 °C
Volume: 10 µl
Substances: Methyl phenyl sulfoxide
Keywords: Chiral separation, Eurocel 02, cellulose based chiral selector
Chromatogram: O
k´1 = 3.77
S CH3
k´2 = 5.35
α = 1.42
0 5 10 15 20 25 30 min

26 Chiral separation of Metoprolol (1-(Isoppropylamino)-
3-[4-(2-Methoxyethyl)Phenoxy]Propan-2-ol)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Heptane / Ethanol (90:10) + 0.1 % DEA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Metoprolol (1-(Isoppropylamino)-3-[4-(2-Methoxyethyl)Phenoxy]Propan-2-ol)
Keywords: beta blocker, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
k´1 = 1.34 NH OH
k´2 = 1.96
α = 1.46
0 5 10 min
V 09/2008 117
27 Chiral separation of 1-(Naphthalene-6-yl)Ethane-1,2-diol
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 1-(Naphthalene-6-yl)Ethane-1,2-diol
Chromatogram:
OH
k´1 = 7.98 OH
k´2 = 6.63
α = 1.20
0 10 20 30 min

28 Chiral separation of Naproxen (6-Methoxy-α-Methyl-2-
Naphthylessigsäure)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Hexan / 2-Propanol (90:10) + 0.1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Naproxen (6-Methoxy-α-Methyl-2-Naphthylessigsäure)
Chromatogram: CH3
k´1 = 0.97 OH
k´2 = 1.17
O
α = 1.21
H3CO
700
600
500
400
300
mAU
200
100
-100
-200
1 2 3 4 5 6 7 8 9 10
Minutes
V 09/2008 119
29 Chiral separation of 1-Phenylethylenglycol
(1-Phenylethane-1,2-diol)
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 1-Phenylethylenglycol (Styrolglycol, 1-Phenylethane-1,2-diol)
Chromatogram:
OH
k´1 = 3.65
OH
k´2 = 3.98
α = 1.09
0 10 20 30 40 min

30 Chiral separation of 1-Phenylpropane-1,2-diol
Method Matrix
Phase: Eurocel 01

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: 1-Phenylpropane-1,2-diol
Chromatogram:
k´1 = 2.76 OH
k´2 = 2.93 OH
α = 1.06
0 10 20 30 min
V 09/2008 121
31 Chiral separation of Pindolol (1-(1H-Indol-4-yloxy)-
3-(1-Methylethylamino)Propan-2-ol)
Method Matrix
Column: Eurocel 01, 5 µm, 125 x 4 mm ID Order No. 12DM370ECJ
Phase: Eurocel 01
Conditions: Eluent: 20 mM Sodium borate buffer / ACN (60:40)

Temperature: 25 °C
Volume: 10 µl
Substances: Pindolol (1-(1H-Indol-4-yloxy)-3-(1-Methylethylamino)Propan-2-ol)
Chromatogram:
HN
k´1 = 1.43
k´2 = 3.68
O NH
α = 2.57
OH
0 3 6 9 min

32 Chiral separation of Pindolol (1-(1H-Indol-4-yloxy)-
3-(1-Methylethylamino)Propan-2-ol)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Acetonitrile + 0.1 % EtOA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Pindolol (1-(1H-Indol-4-yloxy)-3-(1-Methylethylamino)Propan-2-ol)
Chromatogram:
HN
k´1 = 0.6
k´2 = 1.37
α = 2.28 O NH
OH
0 3 6 9 12 min
V 09/2008 123
33 Chiral separation of Propranolol (1-Isopropylamino-
3-(1-Naphthyloxy)-2-Propanol)
Method Matrix
Column: Eurocel 01, 5 µm, 125 x 4 mm ID Order No. 12DM370ECJ
Phase: Eurocel 01
Conditions: Eluent: Acetonitrile / Water (32:68) + 0.1 % DEA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Propranolol (1-Isopropylamino-3-(1-Naphthyloxy)-2-Propanol)
Chromatogram:
k´1 = 12.8
O NH
k´2 = 14.32
OH
α = 1.12
0 10 20 30 min

34 Chiral separation of Propranolol (1-Isopropylamino-
3-(1-Naphthyloxy)-2-Propanol)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: n-Heptane / 2-Propanole (80:20) + 0.1 Ethanolamine

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Propranolol (1-Isopropylamino-3-(1-Naphthyloxy)-2-Propanol)
Chromatogram:
k´1 = 1.75
O NH
k´2 = 2.49
OH
α = 1.42
600
500
400
300
mAU
200
100
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Minutes
V 09/2008 125
35 Chiral separation of Warfarin (4-Hydroxy-3-(3-Oxo-
1-Phenyl-Butyl)-Cumarin)
Method Matrix
Phase: Eurocel 01
Conditions: Eluent: Acetonitrile / Water (50:50) + 0.1 % TFA

Gradient: isocratic
Temperature: 25 °C
Volume: 10 µl
Substances: Warfarin (4-Hydroxy-3-(3-Oxo-1-Phenyl-Butyl)-Cumarin)
Chromatogram:
O O
k´1 = 2.88
k´2 = 4.39
α = 1.53
OH
Eurocel 01: Messung von Warfarin (RP) in saurer mobiler Phase - Block 1
1000
O
800
11,583
600
15,983
400
200
-200
0 2 4 6 8 10 12 14 16 18
Minutes
126 (HPLC)
Chiral Applications
(SFC)
Page
Chiral separation of 1,1´-Bi-2-Naphthol (BINOL) 129
Chiral separation of Ethyl-Mandelate 130
Chiral separation of Hydrobenzoin (1,2-Diphenylethane-1,2-diol) 131
Chiral separation of trans Stilbene Oxide 132
Chiral separation of Warfarin (4-Hydroxy-3-(3-Oxo-1-Phenyl-Butyl)-Cumarin) 133
127
3-Phenylpropionate)
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01
Conditions: Eluent: CO2 + 20 % Methanol

Gradient: isocratic
Flow rate: 2.4 ml/min CO2 / 0.6 ml/min Modifier (Backpressure: 150 bar)
Temperature: 40 °C
Volume: 10 µl
Keywords: SFC, chiral separation, Eurocel 01, cellulose based chiral selector
Chromatogram:
OH
tR1 = 2.23 min
O
tR2 = 2.96 min
0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 min
128 Chiral Applications

2 Chiral separation of 1,1´-Bi-2-Naphthol (BINOL)
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: 1,1´-Bi-2-Naphthol (BINOL), 1,1´-Binaphthalene-2,2´-diol
Chromatogram:
tR1 = 5.19 min
tR2 = 5.71 min

OH
OH
0 1.0 2.0 3.0 4.0 5.0 6.0 min
V 09/2008 129
3 Chiral separation of Ethyl-Mandelate
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: Ethyl-Mandelate
Chromatogram:
tR1 = 1.71 min OH
tR2 = 2.04 min

COOEt
0 1.0 2.0 3.0 4.0 5.0 min

4 Chiral separation of Hydrobenzoin (1,2-Diphenylethane-
1,2-diol)
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: Hydrobenzoin (1,2-Diphenylethane-1,2-diol)
Chromatogram:
OH
tR1 = 2.95 min
tR2 = 3.24 min
OH
0 1.0 2.0 3.0 min
V 09/2008 131
5 Chiral separation of trans Stilbene Oxide
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: trans Stilbene Oxide
Chromatogram:
tR1 = 2.12 min
tR2 = 2.52 min O
0 1.0 2.0 3.0 min

6 Chiral separation of Warfarin (4-Hydroxy-3-(3-Oxo-
1-Phenyl-Butyl)-Cumarin)
Method Matrix
SFC SFC-Mode
Phase: Eurocel 01

Gradient: isocratic
Temperature: 40 °C
Volume: 10 µl
Substances: Warfarin (4-Hydroxy-3-(3-Oxo-1-Phenyl-Butyl)-Cumarin)
Chromatogram: O O
tR1 = 3.26 min
tR2 = 4.98 min

OH
0 1.0 2.0 3.0 4.0 5.0 6.0 min
V 09/2008 133
Substance and Keyword Index
1-(1H-Indol-4-yloxy)-3-(1-methylethylamino)- Acenaphthene ................................................... 18, 19
propan-2-ol ................................................ 122, 123 Acenaphthylene ................................................ 18, 19
1-(3-Dimethylaminopropyl)-1-(4-fluorphenyl)- Acesulfam K ............................................................ 28
3H-2-benzofuran-5-carbonitrile ......................... 105 Acetaldehyde ...................................................... 6, 15
1-(Isoppropylamino)-3-[4-(2-methoxyethyl)- Acetic acid................................................... 30, 31, 50
phenoxy]propan-2-ol ......................................... 117 Acetone ............................................................... 6, 15
1-(naphthalene-6-yl)ethane-1,2-diol ..................... 118 Acetyl cystein .......................................................... 60
1,1´-Bi-2-naphthol ................................................. 129 Acetyldeoxynivalenol .............................................. 48
1,1´-Binaphthalene-2,2´-diol ................................. 129 Acids
1,2 Dimethylbenzene .............................................. 26 aromatic............................................................... 89
1,2-Diphenyl-2-(tosylamino)ethanone................... 107 organic..................................................... 50, 51, 60
1,2-Diphenylethane-1,2-diol.................................. 131 Acrolein ............................................................... 6, 15
10-OH-Carbamazepine........................................... 68 Additives........................................................ 5, 28, 39
1-Aza-[6]helicene .................................................... 96 Adipic acid............................................................... 60
1-Benzene-2-naphthene-3-amino propan(1)ol ....... 97 Adrenaline ............................................................... 71
1-Isopropylamino-3-(1-naphthyloxy)-2-propanol.. 124, Aflatoxin .................................................................. 29
125 Alanine .............................................................. 34, 63
1-Methylnaphthalene .............................................. 18 Alcohols................................................................... 30
1-Phenylethane-1,2-diol........................................ 120 Aldehydes ........................................................... 6, 15
1-Phenylethylenglycol ........................................... 120 Aldicarb ................................................................... 10
1-Phenylpropane-1,2-diol...................................... 121 sulphone .............................................................. 10
2-(4-Bromphenyl)-2-amino acetamid .................... 103 sulphoxide ........................................................... 10
2,2´-bis(diphenylphosphino)- Alkaloids.................................................................. 61
3,3´-bibenzo[b]thiophene................................... 102 Amineptine .............................................................. 62
2,2´-bis(diphenylphosphinoxid)- Amines ............................................ 32, 33, 34, 37, 64
3,3´-bibenzo[b]thiophene................................... 101 Amino acids............................................................. 63
2,4,5-Trichlorophenoxy acetic acid ......................... 22 Amino-dinitrotoluene ............................................... 13
2,4-Dichlorophenoxy acetic acid............................. 22 Amino-nitrotoluene.................................................. 13
2,6-Di-tert.-butyl-4-methylphenol .............................. 5 Aminophenol ........................................................... 79
2-[(4-chlorophenyl)-pyridin-2-yl-methoxy]- Amitriptyline............................................................. 88
N,N-dimethyl-ethanamine.................................. 104 Ammonia ................................................................. 32
2-[4-[2-Hydroxy-3-(1-methylethylamino)propoxy]- Ammonium .............................................................. 11
phenyl]ethanamid ................................................ 94 Ampicillin ................................................................. 80
2-Dimethylamino-1-phenyl-propanol ........................ 114 Angiotensin ............................................................. 81
2-Methylnaphthalene .............................................. 18 Anions
2-Phenyl-1,4-Benzopyrone ................................... 110 inorganic ................................................ 8, 9, 35, 49
3-Amino-2(2-naphtyl)-1-phenylpropanol ................. 93 organic........................................................... 35, 73
4-(2,4-Dichlorophenoxy)butyric acid ....................... 22 Anthocyanidin-3-glycosides .................................... 36
4-(4-Chloro-o-tolyloxy)butyric acid.......................... 22 Anthocyanins........................................................... 36
4,6-Dinitro-o-cresol ................................................. 22 Anthracene........................................................ 18, 19
4-Bromphenyl-glycinamid ..................................... 103 Antibiotics........................................ 65, 66, 77, 80, 87
4-Chloro-o-tolyloxy acetic acid................................ 22 Anticonvulsants ....................................................... 67
4-Hydroxy-3-(3-oxo-1-phenyl-butyl)-cumarin 126, 133 Antidepressant ........................................................ 88
4-Isopropyl-3-methylphenol ...................................... 5 Antidepressants ...................................................... 62
4-oxo-2-cyclohexen-1-yl)-3-methyl- Antiepileptics ........................................................... 68
2,4-pentanedienoic acid ...................................... 92 Antirheumatics ........................................................ 69
6-Methoxy-α-methyl-2-naphthylessigsäure .......... 119 Arginin ..................................................................... 34
6-Nitro-Benzo(a)pyrene .......................................... 16 Arginine ................................................................... 63
9-Methylanthracene ................................................ 26 aromatic acids ......................................................... 89
Abscisic acid ........................................................... 92 Ascorbic acid......................................... 28, 50, 55, 60
134 Substance and Keyword Index

Asparagin.................................................................34 Chloride......................................................8, 9, 35, 49
Aspartame .........................................................28, 39 Chloroaniline............................................................64
Aspartic acid ......................................................34, 63 Chloromethylphenol.................................................23
Atenolol....................................................................94 Chlorophenol ...........................................................23
Atrazine....................................................................21 Chlortoluron .............................................................21
Atropine ...................................................................95 Cholesterol...............................................................54
Barbital.....................................................................83 Chrysene............................................................18, 19
Barium .....................................................................12 cis-Resveratrol.........................................................42
BDMC ......................................................................10 Citalopram .............................................................105
Bentazone................................................................22 Citrate ......................................................................73
Benzo.................................................................18, 19 Citric acid .....................................................31, 50, 60
Benzo(a)pyrene .................................................18, 19 Clopyralid .................................................................22
Benzo(b)fluoranthene ........................................18, 19 Codein......................................................................74
Benzo(e)pyrene .......................................................18 Codeine....................................................................61
Benzo(g,h,i)perylene..........................................18, 19 Corticosterone .........................................................84
Benzo(k)fluoranthene ........................................18, 19 Cortisone............................................................84, 85
Benzoate..................................................................39 Coumarylated anthocyanidin-3-glycosides..............36
Benzoic acid ......................................................28, 53 Cryptopine ...............................................................61
Benzoin....................................................................98 Cyanazin ..................................................................21
Benzyl nicotinate........................................................5 Cyanidol-3-glucosid .................................................36
Benzyl-2-hydroxy-3-phenylpropionate.....99, 100, 128 Cyanocobalamine ....................................................55
Benzyl-Mandelate ....................................99, 100, 128 Cyclohexanone ....................................................6, 15
BINOL ....................................................................129 Cystein .....................................................................60
Biogenic amines ......................................................89 Deoxynivalenol ............................................46, 47, 48
BITIANP.................................................................102 Desethylatrazine ......................................................21
Bradykinin ................................................................81 Desoxycorticosterone ..............................................84
Bromate .....................................................................8 Diaminobutane.........................................................32
Bromide .................................................................8, 9 Diaminopentane.......................................................32
Bromoxynil ...............................................................22 Diaminopropane ......................................................32
BSA..........................................................................82 di-aromatic compounds ...........................................26
Butandiol..................................................................31 Dibenzo(a,h)anthracene ....................................18, 19
Butyl p-hydroxybenzoate ...........................................5 Dibenzothiphene......................................................26
Butylamine ...............................................................32 Dicamba...................................................................22
Butylparabene..........................................................20 Dichlorophenol.........................................................23
Butyraldehyde......................................................6, 15 Dichlorprop ..............................................................22
Cadaverine ........................................................33, 37 Dicloxacillin ..............................................................80
Caempferol ..............................................................42 Diethylamine ............................................................32
Caffeine .............................................................28, 39 Digitalis Glycosides..................................................75
Calcium....................................................................12 Digitoxigenin ............................................................75
Campesterol ............................................................54 Digitoxigenin bisdigitoxoside ...................................75
Camphor ..................................................................69 Digitoxigenin monodigitoxoside ...............................75
Carbamates .............................................................10 Digitoxin ...................................................................75
Carbamazepine .................................................67, 68 Digoxigenin ..............................................................75
Carbamazepine-diol.................................................68 Digoxigenin bisdigitoxoside .....................................75
Carbamazepine-epoxide .........................................68 Digoxigenin monodigitoxoside.................................75
Carbaryl ...................................................................10 Digoxin .....................................................................75
Carbinoxamine.......................................................104 Dihydrjoxybenzylamine............................................71
Carbofuran...............................................................10 Dihydroxyphenylacetic acid .....................................89
Carbohydrates .........................................................30 Diketopiperazine ......................................................39
Carbonyl compounds...........................................6, 15 Dilactid ...................................................................106
Carotene-β.........................................................38, 58 Dimethylamine .........................................................32
Catecholamines .................................................70, 71 Dimethylphenol ........................................................23
Cations Dinitrobenzene.........................................................13
divalent.................................................................12 Dinitrophenol............................................................23
monovalent...........................................................11 Dinitrotoluene...........................................................13
Chloracetanilide .......................................................79 Dinoterb ...................................................................22
Chloramphenicol......................................................72 Diosmin ....................................................................43
Chlorate .....................................................................9 Di-tert.-butylmethylphenol..........................................5
V 09/2008 135
Diuron...................................................................... 21 Hexahydro-6,6-dimethyl-2-tosylpyrrolo
DNOC...................................................................... 22 [1,2-e]imidazol-3-one......................................... 111
Dopamine................................................................ 71 Hexamethylbenzene ............................................... 26
Doxepin ................................................................... 88 Hexazinon ............................................................... 21
DPH......................................................................... 68 Hexogen.................................................................. 13
Drugs....................................................................... 78 Hexylamine ............................................................. 32
Antibacterial......................................................... 72 Histamine .................................................... 32, 33, 37
Antibiotics .......................................... 65, 66, 77, 80 Histidine ............................................................ 34, 63
Anticonvulsant ..................................................... 67 HMF ........................................................................ 44
Antidepressant .............................................. 62, 88 Homovanillic acid .................................................... 89
Antiepileptics ....................................................... 68 Hydrobenzoin ........................................................ 131
Antirheumatics..................................................... 69 Hydroxy-benzoic acid.............................................. 53
Glycosides........................................................... 75 Hydroxycarbofuran.................................................. 10
Narcotics ............................................................. 76 Hydroxymethylfurfural (HMF).................................. 44
Eledoisin.................................................................. 81 Hydroxyphenyl ethanone ........................................ 79
Elphinidol-3-glucosid............................................... 36 Hydroxyphenyl ethanone oxime.............................. 79
Ergosterol................................................................ 54 Hydroxypyrene ........................................................ 14
Eriocitrin .................................................................. 43 Hydroxyzine .......................................................... 112
Eriodictyol 7-O-glucoside ........................................ 43 Imipramine .............................................................. 88
Esteriol .................................................................... 85 Indeno(1,2,3-cd)pyren....................................... 18, 19
Estradiol .................................................................. 85 Indole ...................................................................... 32
Estrone.................................................................... 85 Insecticides ............................................................. 10
Ethanol .............................................................. 30, 31 Insulin bovine .......................................................... 82
Ethanolamine .......................................................... 32 Iodate ........................................................................ 9
Ethosuximide .......................................................... 68 Iodide ........................................................................ 9
Ethyl(3-methyl-4-oxo-5-piperidinothiazolidin- Ioxynil ...................................................................... 22
2-ylidene)acetate............................................... 109 Isoamylamine .......................................................... 33
Ethylamine .............................................................. 32 Isoleucine .......................................................... 34, 63
Ethylenglycolmonosalicylate ................................... 69 Isopentylamine ........................................................ 32
Ethyl-Mandelate ............................................ 108, 130 Isopropylamine........................................................ 32
Ethylparabene......................................................... 20 Isopropylmethylphenol .............................................. 5
Etozolin ................................................................. 109 Isoproturon .............................................................. 21
Explosives ............................................................... 13 Isorhoifolin............................................................... 43
Fenoxyprop-p.......................................................... 22 Ketones ............................................................... 6, 15
Flavonoid glycosides............................................... 43 Lactic acid ..................................... 30, 31, 50, 51, 113
Flavonoids......................................................... 41, 42 Lactose.................................................................... 45
Flavonone ............................................................. 110 Lanosterol ............................................................... 54
Fluazifop-p .............................................................. 22 Laurylalcohol ........................................................... 24
Fluoranthene..................................................... 18, 19 polyglycol ether ................................................... 24
Fluorene ...................................................... 18, 19, 26 Leucine.............................................................. 34, 63
Fluoride ..................................................................... 8 Linuron .................................................................... 21
Fluroxypyr ............................................................... 22 Lithium..................................................................... 11
Folic acid ................................................................. 55 Luminal.............................................................. 76, 83
Formaldehyde ..................................................... 6, 15 Luteolin 7-O-rutinoside............................................ 43
Formic acid.............................................................. 30 Lysine................................................................ 34, 63
Fructose ............................................................ 31, 45 Lysozyme ................................................................ 82
Fumaric acid ........................................................... 50 Magnesium.............................................................. 12
Furfural.................................................................... 44 Malic acid .................................................... 31, 50, 51
Fusarium mycotoxines ............................................ 48 Malonic acid ............................................................ 50
Glucose ....................................................... 30, 31, 45 Maltose.............................................................. 30, 45
Glutamic acid .................................................... 34, 63 Malvidol-3-glucosid ................................................. 36
Glutamin............................................................ 34, 63 MCPA...................................................................... 22
Glycine .............................................................. 34, 63 MCPB...................................................................... 22
Glycosides............................................................... 75 Mecoprop ................................................................ 22
Guaifenesin ............................................................. 74 Mesotrione .............................................................. 22
Haloxyfop ................................................................ 22 Metazachlor............................................................. 21
Heptylamine ............................................................ 33 Methabenzthiazuron ............................................... 21
Hesperidin ......................................................... 41, 43 Methacrolein........................................................ 6, 15

Methanol ..................................................................30 Nitro-Pyrene.............................................................16
Methiocarb ...............................................................10 Nitrotoluene .............................................................13
Methionine .........................................................34, 63 Nivalenol ............................................................47, 48
Methomyl .................................................................10 N-Methylephedrin ..................................................114
Methoxy benzoic acid ..............................................53 Nonylphenol polyglycol ether...................................24
Methyl ethyl ketone..............................................6, 15 Noradrenaline ..........................................................71
Methyl phenyl sulfoxide .................................115, 116 Norgestrel ................................................................84
Methyl p-hydroxybenzoate ........................................5 Nortiptyline...............................................................88
Methyl-1H-tetrazole-5-thiol ......................................77 N-phenylacetamide..................................................79
Methyl-6α-hydroxy-11β-progesterone .....................85 Octogen ...................................................................13
Methyl-6α-hydroxy-17α-progesterone.....................85 Octopamine .............................................................32
Methyl-6α-hydroxy-17α-progesterone acetate ........85 Octyl hydrogen sulphate ............................................7
Methylamine ............................................................32 Oleic acid diethanolamide........................................24
Methylbutylamine.....................................................32 Oleic amide polyglycol ether....................................24
Methyldinitrophenol..................................................23 Organic acids.....................................................30, 31
Methylparabene .......................................................20 Ovalbumin................................................................82
Metobromuron .........................................................21 Oxalate...............................................................35, 73
Metolachlor ..............................................................21 Oxalic acid ...............................................................50
Metoprolol ..............................................................117 Oxamyl.....................................................................10
Metoxuron................................................................21 Oxcarbazepine.........................................................68
Monoammonium glycyrrhizinate................................5 Oxytocin ...................................................................81
mono-aromatic compounds .....................................26 PAH..........................................................................19
Monolinuron .............................................................21 Palmitic acid polyglycol ester...................................24
Morphine..................................................................61 Pamidronate disodium .............................................78
Morpholine ...............................................................32 Pantothenylethylether ................................................5
Moxalactam .............................................................77 Papaverine...............................................................61
Mycotoxins...................................................46, 47, 52 Parabenes ...............................................................20
Myoglobin ................................................................82 Paracetamol.............................................................79
Myrcetine .................................................................42 Patulin ......................................................................52
Myristylalcohol .........................................................24 PEMA.......................................................................68
Myristylalcohol polyglycol ether ...............................24 Penicillin...................................................................80
N-(2-hydroxyphenyl)acetamide ...............................79 Pentachlorphenol.....................................................23
N-(4-hydroxyphenyl)propanamide ...........................79 Pentanone ...........................................................6, 15
Nafcillin ....................................................................80 Pentylamine .............................................................32
Naphthalene ................................................18, 19, 26 Pentyl-i-amine..........................................................33
Naphtol ....................................................................10 Peonidol-3-glucosid .................................................36
Naproxen ...............................................................119 Peptides ...................................................................81
Narcotics..................................................................76 Pesticides ................................................................21
Narcotine .................................................................61 Petunidol-3-glucosid ................................................36
Naringin ...................................................................41 Phenanthrene ..............................................18, 19, 26
Narirutin ...................................................................43 Phenethylamine .................................................32, 33
Neurotensin .............................................................81 Phenobarbital.....................................................67, 68
Nicosulfurone...........................................................22 Phenol......................................................................23
Nicotinamide ............................................................55 Phenols ....................................................................23
Nicotinic acid............................................................55 Phenylalanine ....................................................34, 63
Nicotinic acid benzyl ester .......................................69 Phenyldodecane ......................................................26
Nicotinic acid methyl ester.......................................69 Phenylene diamine ..................................................64
Nitrate ........................................................8, 9, 35, 49 Phenylethylamine ....................................................37
Nitrite....................................................................8, 49 Phenytoin .................................................................67
Nitroaniline...............................................................17 Phosphate............................................................8, 35
Nitrobenzene ...........................................................13 p-Hydroxy butylbenzoate .........................................53
Nitro-Benzo(a)pyrene ..............................................16 p-Hydroxy ethylbenzoate .........................................53
Nitro-Chrysene.........................................................16 p-Hydroxy methylbenzoate ......................................53
Nitro-Fluorene..........................................................16 p-Hydroxy propylbenzoate.......................................53
Nitro-Naphthalene....................................................16 Pindolol ..........................................................122, 123
Nitro-PAH.................................................................16 Piperidine .................................................................32
Nitro-Phenanthrene .................................................16 Plant hormone .........................................................92
Nitrophenol ........................................................23, 79 Polyethylene glycol ..................................................24
V 09/2008 137
Polyoxyethylene Sulcotrione .............................................................. 22
glycerol monostearate ......................................... 24 Sulfa drugs ........................................................ 86, 87
sorbitane monolaurate......................................... 24 Sulfadiazine....................................................... 86, 87
Potassium ............................................................... 11 Sulfadimethoxine .................................................... 87
Prednisolone ........................................................... 85 Sulfamerazine ......................................................... 86
Primidone .......................................................... 67, 68 Sulfamethazine ....................................................... 86
Progesterone .................................................... 84, 85 Sulfamethiazole....................................................... 87
Prominal ............................................................ 76, 83 Sulfamethoxazol...................................................... 66
Propionaldehyde ................................................. 6, 15 Sulfamethoxazole ................................................... 87
Propoxur.................................................................. 10 Sulfamethoxydiazine............................................... 65
Propranolol.................................................... 124, 125 Sulfamethoxypyridazine.......................................... 87
Propylamine ............................................................ 32 Sulfanilamide..................................................... 65, 86
Propylparabene....................................................... 20 Sulfate ............................................................. 8, 9, 35
Proteins ................................................................... 82 Sulfathiazole............................................................ 86
Pseudoefedrin......................................................... 74 Sulpho fatty acid-α-methyl esters ........................... 25
Putrescine ......................................................... 33, 37 Sulphosuccinate........................................................ 7
Pyrene............................................................... 18, 19 Surfactants .................................................... 7, 24, 25
Pyridoxale ............................................................... 55 Sweeteners ....................................................... 28, 39
Pyridoxamine .......................................................... 55 Tartaric acid ...................................................... 31, 50
Pyridoxine ............................................................... 55 Terbuthylazine......................................................... 21
Pyridoxine dioctanoate.............................................. 5 Testosterone ..................................................... 84, 85
Pyrrolidine ............................................................... 32 Tetryl ....................................................................... 13
Pyrrolidone .............................................................. 32 Thebaine ................................................................. 61
Quercetine............................................................... 42 Thiamine ................................................................. 55
Quinic acid ........................................................ 50, 51 Thiogenal .......................................................... 76, 83
Quinine.................................................................... 28 Threonine .......................................................... 34, 63
Quinmerac............................................................... 22 Tocopherols ............................................................ 56
Revinal .................................................................... 70 Tocopherol-α........................................................... 58
Revonal ............................................................. 76, 83 Tocopheryl acetate ................................................... 5
Rhodanide................................................................. 9 Toluidine-o .............................................................. 65
Riboflavine .............................................................. 55 trans Stilbene oxide .............................................. 132
Ribonuclease A....................................................... 82 trans-Resveratrol..................................................... 42
Rosmarinic acid ...................................................... 43 tri-aromatic compounds .......................................... 26
Saccharides ............................................................ 45 Trichlorocarbanilide................................................... 5
Saccharin ................................................................ 28 Trichlorphenol ......................................................... 23
Saccharose ............................................................. 45 Trichotecenes.......................................................... 46
Salicylic acid ............................................................. 5 Trichothecens.......................................................... 47
Sebuthylazin ........................................................... 21 Trimethoprim ........................................................... 66
Sedativa .................................................................. 83 Trimipramine ........................................................... 88
Serine................................................................ 34, 63 Tryptamine .............................................................. 37
Serotonin................................................................. 32 Tryptophan .............................................................. 89
Shikimic acid ..................................................... 50, 51 Tyramine ................................................................. 37
Simazin ................................................................... 21 Tyrosine ............................................................ 34, 63
Sitosterol ................................................................. 54 Valine ................................................................ 34, 63
Sodium .................................................................... 11 Vanilin ..................................................................... 28
Sodium soap (C8 - C18) ......................................... 25 Veronal.................................................................... 76
Sorbic acid ........................................................ 28, 53 Vitamin A acetate .............................................. 57, 58
Stearic acid polyglycol ester ................................... 24 Vitamin A alcohol .................................................... 57
Stearyl glycyrrhetinate .............................................. 5 Vitamin D2......................................................... 56, 57
Steroids ....................................................... 54, 84, 85 Vitamin D3............................................................... 57
Steroles ................................................................... 54 Vitamin E................................................................. 57
Stigmasterol ............................................................ 54 Vitamin K1............................................................... 57
Stilbene ................................................................... 42 Vitamins .......................................... 38, 55, 56, 57, 58
Strontium................................................................. 12 Warfarin......................................................... 126, 133
Styrolglycol............................................................ 120 α-Hydroxy-α-phenylacetophenon ........................... 98
Succinic acid ............................................... 30, 31, 50

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This Applications Journal is also available as an always actualized

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KNAUER - HPLC, SMB, Osmometry

Laboratory Instrumentation and Separation Solutions
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Applications Journal

CHIRAL APPLICATIONS 127

SUBSTANCE AND KEYWORD INDEX 134

This Applications Journal was designed to provide you with detailed

KNAUER - HPLC, SMB, Osmometry

Column: Eurospher 100-5 C18, 250 x 4 mm ID Order No. 25DE181ESJ

Phase: Eurospher 100-5 C18

Substances: Benzyl nicotinate, Butyl p-hydroxybenzoate, 2,6-Di-tert.-butyl-4-methylphenol, 4-

Column: Eurospher 100-5 C18, 250 x 4 mm ID Order No. 25DE181ESJ

Phase: Eurospher 100-5 C18

Conditions: Eluent: Methanol / Water (84:16)

Substances: DNPH derivates of Formaldehyde, Acetaldehyde, Acrolein, Propionaldehyde,

Keywords: Aldehydes, Carbonyl compounds, Ketones

Column: Eurospher 100-5 C18, 250 x 4 mm ID Order No. 25DE181ESJ

Phase: Eurospher 100-5 C18

Conditions: Eluent: 0.01 M aqueous Tetrabutylammonium Sulphate / Methanol (23:77),

Detection: Refractive Index

Substances: Octyl Hydrogen Sulphate, C10 Sulphosuccinate,

Column: Novosep A-2 Anion, 5 µm, 250 x 4 mm ID Order No. B92

Phase: Novosep A-2 Anion

Conditions: Eluent: 3.6 mM Sodium Carbonate (Na2CO3)

Detection: suppressed Conductivity (Alltech Model 650), Range 0.5 µS

Substances: Fluoride, Bromate, Chloride, Nitrite, Bromide, Nitrate, Phosphate, Sulfate

Column: Eurospher 100-5 C18, 250 x 4.6 mm ID Order No. 25EE181ESJ

Phase: Eurospher 100-5 C18

Conditions: Eluent: 0.11 % Acetyl trimethylammonium p-toluenesulphonate +

Detection: UV at 236 nm and 254 nm,

Substances: Cl-, ClO3-, SO42-, NO3-, I-, IO3-, SCN-, Br-

Column: Eurospher 100-5 C18, 250 x 4.6 mm ID Order No. 25EE181ESJ

Phase: Eurospher 100-5 C18

Conditions: Eluent: A: Water B: Methanol

Substances: Aldicarb, Aldicarb sulphoxide, Aldicarb sulphone, BDMC, Carbaryl, Carbofuran, 3-

Keywords: Carbamates, Insecticides

Phase: PRP-X 200, cation exchange resin

Conditions: Eluent: 4 mM Nitric Acid in 30 % MeOH

Detection: Conductivity (negative Polarity)

Substances: Ammonium, Lithium, Potassium, Sodium

Keywords: Cations, monovalent

1. Lithium (5.0 ppm)

Phase: PRP-X 200, cation exchange resin

Conditions: Eluent: 1.5 mM Ethylene Diammonium Dichloride

Detection: Conductivity (negative Polarity)

Substances: Barium, Calcium, Magnesium, Strontium

Keywords: Cations, divalent

1. Magnesium (20.0 ppm)

Column: Ultra Sep ES EX, 250 x 3 mm ID Order No. I0034

Conditions: Eluent: A: Methanol B: Water

Substances: 2-Amino-4,6-dinitrotoluene, 4-Amino-2,6-dinitrotoluene, 2-Amino-4-nitrotoluene, 2-

Keywords: Explosives, EPA 8330

Column: Eurospher 100-5 C18, 250 x 4 mm Order No. 25DE181ESJ

Phase: Eurospher 100-5 C18

Conditions: Eluent: A: ACN B: H2O

Detection: Fluorescence: excitation 242 nm, emission 388 nm

Substances: Hydroxypyrene, Biomarker of PAH contamination

Column: ProntoSIL 120-3 C18 AQ, 250 x 3 mm ID Order No. 15DF080PSJ

Phase: ProntoSIL 120-3 C18 AQ

Conditions: Eluent: A: Water B: Acetonitrile

Substances: DNPH derivates of Formaldehyde, Acetaldehyde, Acrolein, Propionaldehyde,

Keywords: Aldehydes, Carbonyl compounds, Ketones