QUALITATIVE TESTS SUMMARY v3

(FOR MODULE 2 OF THE PH PHARMACIST LICENSURE EXAMS)

Made by: Ser Loisse Mortel

NOTES:
1) This material is allowed for use by anyone and is NOT linked or affiliated to any institution.
2) This material is NOT for sale. The author has the right to pursue individuals who lift or outright copy
the pages of this material, especially if used for their financial benefit.
3) Some information may vary from source to source. Additionally, some of these tests are obsolete or
rarely done in the modern day. The author accepts potential inaccuracies and encourages users to fact-
check and perform any necessary corrections.

BIOCHEMISTRY
A. PROTEINS/AMINO ACIDS
NAME OF TEST FOR REAGENT POSITIVE
RESULT
1. Ninhydrin Free amino acids Ninhydrin Violet
2. Biuret Peptide bonds Cupric sulfate in NaOH Violet
3. Xanthoproteic Aromatic compounds Conc. Nitric acid Yellow
(AA: Phe, Trp, Tyr)
4. Millon’s Phenolic compounds Mercury in nitric acid Rose/salmon
(AA: Tyr)
5. Hopkin’s Cole Indole (AA: Trp) Glyoxylic acid in Violet ring
sulfuric acid
6. Sakaguchi Guanidine group (AA: Arg) 10% alpha-naphthol in Reddish/wine
NaBrO/NaClO
7. Nitroprusside Thiol group (AA: Cys) Sodium nitroprusside Red
8. Fohl’s Sulfur containing Lead II acetate Brown or black ppt
compounds (AA: Cys, Met)
9. Amide Amidic compounds NaOH + heat Litmus paper R->B,
(AA: Asn, Gln) B->B
10. Pauly’s Phenolic and imidazole Diazotized sulfanilic acid Red
groups (AA: Tyr, His)

B. NUCLEIC ACIDS
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Dische Deoxyribose Diphenylamine Blue due to hydroxylevulinic
(DNA) aldehyde
2. Ammonium Phosphate group Ammonium Yellow ppt
molybdate molybdate in acid
3. Murexide Purines (Bases A, Potassium chlorate in Pink color
G HCl
4. Wheeler-Johnson Pyrimidines Bromine, barium Green with bromine (C and U
(bases U, T, C) hydroxide turns into dialuric acid); purple
with barium hydroxide

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C. CARBOHYDRATES
NAME OF TEST FOR REAGENT POSITIVE
RESULT
1. Molisch 10% alpha-naphthol in Violet ring
Carbohydrates, in general ethanol
2. Anthrone Anthrone Dark green
3. Iodine Starch Iodine Violet
4. Benedict’s Cupric ions in alkaline Brick red;
Reducing sugars; medium (Barfoed’s reacts faster
5. Barfoed’s Monosaccharides Cupric ions in acidic to monosaccharides
(Barfoed’s) medium
than more complex
sugars)
6. Seliwanoff’s Ketoses Resorcinol in HCl Cherry red
7. Bial’s Orcinol in FeCl3 and Blue-green
Pentoses HCl
8. Phloroglucinol Phloroglucinol Red
9. Phenylhydrazine Glucose/galactose/fructose Dinitrophenylhydrazine Osazone crystals
(DNP) in Sodium Lac – cottonball-shaped
Mal – sunflower-shaped
Acetate Glu/Fru/Man – needle/
broomstick-shaped
10. Mucic acid (aka Galactose Nitric acid Insoluble crystals
Galactaric acid)

D. LIPIDS
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Sudan III Tetraazobenzene + beta- Red
Fatty acids naphthol in 80% alcohol
(all are rarely
2. Osmic acid Osmium tetroxide Black
done today)
3. Tincture of Alkanna Alkanet root extract Red
4. Kraut’s Choline Potassium bismuth iodide Dark-red ppt
(the same material in
Dragendorff’s reagent)
5. Liebermann- Sterol Sulfuric acid and acetic Green/blue (unsaturated),
Burchard anhydride yellow (saturated)
6. Acrolein Glycerol Heat Acrid odor
7. Bromine Bromine in CCl4 Disappearance of orange
color
* For hydrogenationed vegetable oil = Serger test
* For sesame oil = Baudoin test
* For cottonseed oil = Bevan test
* For olive oil = Millon’s test (to detect contamination by tea tree or camellia)

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PHARMACOGNOSY/ PLANT CHEMISTRY
A. TANNINS
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Goldbeater’s skin Membrane of ox Black/brown
True tannins, in
intestine, HCl, FeSO4
general
2. Gelatin Gelatin TS Gel-like precipitate
3. Ferric chloride Ferric chloride TS Bluish black (hydrolysable),
greenish brown (condensed)
4. Bromine test Bromine reagent Buff ppt. (CONDENSED only)
5. KMnO4 test Distinguishing Potassium Disappearance of violet color
true tannins permanganate (HYDROLYSABLE only)
6. Dry distillation Heat Pyrogallol as product
(hydrolysable), Pyrocatechin
as product (condensed)
7. KCN KCN Reddish-orange color, when
shaken turns into a pinkish
foam
8. Chlorogenic acid Chlorogenic Ammonia Green
acid

B. ALKALOIDS
NAME OF TEST REAGENT POSITIVE RESULT
1. Wagner’s Iodine in KI Reddish brown ppt
(1% in 1% = Bouchardat’s)
2. Valser’s Mecuric Iodide White ppt
3. Mayer’s Mercuric chloride in KI White/ Cream ppt
4. Dragendorff’s Potassium bismuth iodide
Orange ppt
(modified – Kraut’s)
5. Marme’s Potassium cadmium iodide
6. Hager’s Saturated picric acid
7. Scheibler’s Phosphotungstic acid* Precipitation
8. Sonnenschein’s Phosphomolybdic acid*
9. Reaction with metals Gold/silver/platinum salts
* Both acids can be combined in a specific ratio to create the Folin-Ciocalteu reagent for quantifying
polyphenols in a sample

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C. VOLATILE OILS
NAME OF TEST REAGENT POSITIVE RESULT
1. Blot test Paper Grease will disappear; if not, they are fixed oils
(such as fatty acids)
2. Odor test Diverse fragrant odors

D. SAPONINS
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Froth Plant extract itself Honeycomb froth greater
than 2cm for 10 minutes or
more
2. Hemolysis Plant extract on blood agar Zones of hemolysis (aka
hemolytical halos)

3. Capillary Plant extract itself in capillary Reduction of at least ½

tube height of water in capillary
tube
4. Reaction with Barium/ Plumbous ions
Beige to brown complex
metals
5. Liebermann- Nucleus of Sulfuric acid and acetic Green/blue (steroidal),
Burchard saponins anhydride red/pink/violet (triterpenoid)

E. FLAVONOIDS
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Batesmith- Chalcone/ aurone HCl Immediate red (to verify
Metcalf chalcone/aurone)
If no color, boil, and red/violet color
is seen (to verify leucoanthocyanins)
2. Wilstatter Gamma-benzopyrone HCl, Orange -> red -> crimson -> magenta,
Cyanidin nucleus of all Magnesium, ocasionally green or blue
flavonoids Octanol
3. Matchstick Catechins Matchstick in Pink stain on wood
HCl

F. ANTHRAQUINONES
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Borntrager’s Anthraquinones NH4OH Red
2. Modified Anthraquinone NH4OH + FeCl3 and HCl Pink
Borntrager’s glycosides

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G. CARDIAC GLYCOSIDES
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Salkowski Sterol Sulfuric acid Red/violet
2. Liebermann- Unsaturated Sulfuric acid and acetic Green/blue (unsaturated),
Burchard sterol anhydride yellow (saturated)
3. Kedde’s Unsaturated Dinitrobenzoic acid in Blue-violet (or amethyst)
lactone NaOH color
4. Keller-Kiliani Deoxysugar FeCl3 in acetic acid and
Reddish brown
H2SO4

H. ALCOHOL GLYCOSIDES
NAME OF TEST REAGENT POSITIVE RESULT
1. Ester formation Carboxylic acid (usually acetic acid) Fragrant (usually fruity) odor
Specific reactions:
Salicin (from Salix and Populus) on hydrolysis à saligenin (aka salicyl alcohol) + sugar

I. PHENOL GLYCOSIDES
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Millon’s Phenolic groups Mercury in nitric acid Reddish-brown
CHROMATOGRAPHY
2. Folin-Ciocalteu Catechol/hydroquinone Folin-Ciocalteu Reagent Blue spots
(Phosphomolybdate/
Phosphotungstate)
3. Vanillin HCl Resorcinol/phloroglucinol Vanillin HCl Pink spots
4. Gibb’s Gibb’s Reagent (2,6- Various colors
dichloroquinone-4-
chloroimide)
Specific reactions:
Arbutin (from uva ursi) on hydrolysis à hydroquinone + sugar

J. ALDEHYDE GLYCOSIDES
NAME OF TEST REAGENT POSITIVE RESULT
1. Fehling’s Fehling’s A (Cupric sulfate) and Fehling’s B Brick red
(Potassium Sodium Tartrate)
2. Nessler’s Nessler’s Reagent – potassium mercuric Gray
iodide (K2HgI4)
3. Tollen’s Ammoniacal silver nitrate Silver mirror
4. Schiff Decolorized magenta-red pigment Recolorization
Specific reactions:
Glucovanillin (from vanilla pods) on hydrolysis à Vanillin + glucose

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K. LACTONE GLYCOSIDES
NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Kedde’s Unsaturated Dinitrobenzoic acid in NaOH Blue-violet (or amethyst)
lactone color

L. ISOTHIOCYANATE GLYCOSIDES
NAME OF TEST REAGENT POSITIVE RESULT
1. Odor test (for n/a Mustard smell due to SCN
volatiles)
2. Ferric chloride (for Ferric chloride SCN- + Fe+3 -> Fe(SCN)3 (blood red)
nonvolatiles)
Specific reactions:
Sinalbin (from white mustard) + myrosin à Acrinyl isothiocyanate + glucose
Sinigrin (from black mustard) + myrosin à Allyl isothiocyanate (volatile mustard oil) + glucose
Gluconapin (from rapeseed B. napus) + myrosin à But-3-enyl isothiocyanate + glucose
Gluconasturtiin (from watercress) + myrosin à Phenylethyl isothiocyanate + glucose
ID test of garlic: Sodium nitroferricyanide (red/orange)

M. CYANOGENIC GLYCOSIDES
NAME OF TEST REAGENT POSITIVE RESULT
1. Guignard Sodium picrate paper Brick red/ various shades of red due to formation
of sod. purpurate
2. Odor n/a Bitter almond/ peach kernel smell due to HCN
Specific reactions:
Amygdalin (from prunes like almond and peach) + emulsin (an enzyme complex of amygdalase
and prunase) à benzaldehyde + HCN + 2 glucose units overall
- Consists of 3 steps
a) Amygdalin + amygdalase à prunasin + 1 glucose
b) Prunasin + prunase à mandelonitrile + 1 glucose (plus the glucose in step a = 2 glucose)
c) Mandelonitrile + H2O à benzaldehyde + HCN

N. TESTS FOR SPECIFIC CONSTITUENTS (lowest priority table)

NAME OF TEST FOR REAGENT POSITIVE RESULT
1. Baudoin Sesame oil HCl, furfural Pink
2. Duquenois Marijuana Duquenois reagent, HCl, Violet on chloroform layer
chloroform
3. Marquis Codeine/Opioids Marquis reagent Blue
4. Hasemann Ferric chloride Blood red
5. Vitali Atropine/Tropanes Nitric acid Gray ppt.
6. Gulielmo Sulfuric acid Red with flower odor
7. Gerard’s Ethanol, mercuric chloride Yellow

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 8. Thalleioquin Quinine Acetic acid, chlorine water, Fine blue fluorescence,
ammonia then green ppt with
ammonia
9. Herapathite Acetic acid, alcohol, sulfuric Green leaflet with metallic
acid, iodine luster
10. Erythroquinine Acetic acid, bromine water, Brilliant red violet
NH4OH, K ferricyanide, CH3I
11. Mandelin Strychnine Mandelin reagent Violet
12. Amalic acid Caffeine Chlorine, NH4OH Reddish brown with
chlorine
Transient purple with
NH4OH
13. Carr Price Vitamin A SbCl3 Blue to green to pink to
gray
14. Furter-Meyer Vitamin E N-butanol, HNO3 Bronze red

ADDITIONAL NOTES
WATER-SOLUBLE VITAMINS (excluding defunct/former vitamins)
Vitamins with multiple forms (e.g. D, K) are represented by the common version in humans
Code Name Cofactor Primary functions Deficiency
C Ascorbic acid Ascorbic acid (itself) Collagen synthesis Scurvy
(pro/lys hydroxylation)
B1 Thiamine/ Thiamine pyrophosphate (TPP) Aldehyde transfer Beriberi
Aneurine Cell respiration
B2 Riboflavin Flavin mononucleotide (FMN) Redox Cheilosis, glossitis
Flavin adenine dinuclotide (FAD) (mainly as carriers for the
B3 Niacin/ Nicotinamide adenine dinucleotide electron transport chain) Pellagra
Nicotinic acid (NAD)
B5 Pantothenic acid Coenzyme A (CoA) Anything CoA Burning foot syndrome
(fatty acid/sterol
synthesis, Krebs’ cycle)
B6 Pyridoxine Pyridoxal phosphate (PLP) Transamination Peripheral neuropathy
B7/H Biotin Biotin (itself) Carboxylation Hair thinning, brittle nails
B9 Folic acid Tetrahydrofolate (THF) 1-carbon transfers, Megaloblastic anemia,
nucleotide synthesis neural tube defects
B12 Cobalamin Deoxyadenosylcobalamin (DAC) Methylation, fatty acid Megaloblastic anemia,
and amino acid metab neuropathy
A Retinoids Retinol (for transport) Light sensitivity (retinal) Nyctalopia (night
Retinal (binds with rhodopsin) Antiacne/teratogenicity blindness), xerophthalmia
Retinoic acid (affects cell growth) (retinoic acid derivatives)
D3 Cholecalciferol 1,25-dihydroxycalciferol (calcitriol) Ca absorption Osteolamacia, rickets
E Tocopherols Tocopherols Antioxidant CNS/muscle weakness,
ataxia
K1 Phytomenadione/ Reduced vitamin K (KH2) Coagulation Bleeding
Phylloquinone (But the epoxide form K>O is the substrate (activation of vitamin K-
of VKOR (target enzyme of warfarin)) dependent factors)

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 SAMPUNG HALAMANG GAMOT
Bayabas Psidium guajava Astringent
Ampalaya Momordica charantia Antidiabetic
Bawang Allium sativum Antidyslipidemic
Yerba Buena Mentha cordifolia Analgesic
(aka Clinopodium douglasii)
Pansit-pansitan Peperomia pellucida Anti-gout
Lagundi Vitex negundo Anti-asthma
Akapulko Cassia alata Antifungal
Niyog-niyogan Quisqualis indica Anthelminthic
Tsaang Gubat Carmona retusa Antidiarrheal
(aka Ehretia microphylla)
Sambong Blumea balsamifera Diuretic

MAIN PLANT GROWTH REGULATORS/ HORMONES

Name Effects (simplified)
Auxin • Apical dominance
• Tropisms (gravi-, photo-)
• Stem elongation
Gibberelin • Germination
• Subapical elongation
• Stem elongation
Cytokinin • Cell division
• Prevention of senescence
Ethylene • Fruit ripening
• Abscission (primary)
Abscisic acid • Dormancy and inactivity (prevents germination, ripening,
and photosynthesis)
• Abscission (rare)

References:
1. Crisostomo, A.C. et. al. (2010). Laboratory Manual in General Biochemistry. University of
Santo
Tomas: CnE Publishing, Inc.
2. Laboratory Manual in Pharmaceutical Chemistry 2. University of Santo Tomas.
3. Tyler, V., Brady, L., Robbers, J. (1988). Pharmacognosy 9th Edition. Philadelphia: Lea &
Febiger.
4. Evans, W.C. (2002). Trease and Evans Pharmacognosy. Amsterdam: Elsever B.V.
+ Various internet sources and powerpoints of Prof. Rebecca David

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