Isoprenaline hydrochloride EUROPEAN PHARMACOPOEIA 11.

Time Mobile phase A Mobile phase B

(min) (per cent V/V) (per cent V/V)
0 - 12 100 0

12 - 20 100 → 85 0 → 15
C. pyridine-4-carbonitrile (isonicotinonitrile),
20 - 28 85 15

Flow rate : 1.5 mL/min.

Detection : spectrophotometer at 266 nm.
Injection : 10 μL.
Identification of impurities : use the chromatogram obtained D. pyridine-3-carbohydrazide (nicotinoyl hydrazide),
with reference solution (b) to identify the peaks due to
impurities A, B and D.
Relative retention with reference to isoniazid (retention E. hydrazine.
time = about 8.4 min): impurity A = about 0.5 ;
impurity D = about 1.15 ; impurity B = about 1.4. 04/2020:1332
System suitability : reference solution (b) :
– peak-to-valley ratio : minimum 1.8, where Hp = height
above the baseline of the peak due to impurity D and
Hv = height above the baseline of the lowest point of the
curve separating this peak from the peak due to isoniazid. ISOPRENALINE HYDROCHLORIDE
Calculation of percentage contents :
– correction factors : multiply the peak areas of the following Isoprenalini hydrochloridum
impurities by the corresponding correction factor :
impurity A = 1.4 ; impurity B = 1.5 ;
– for each impurity, use the concentration of isoniazid in
reference solution (a).
Limits:
– impurities A, B : for each impurity, maximum 0.15 per cent ; C11H18ClNO3 Mr 247.7
– unspecified impurities : for each impurity, maximum [51-30-9]
0.10 per cent ; DEFINITION
– total : maximum 0.5 per cent ; (1RS)-1-(3,4-Dihydroxyphenyl)-2-[(1-methylethyl)amino]-
– reporting threshold : 0.05 per cent. ethanol hydrochloride.
Loss on drying (2.2.32) : maximum 0.5 per cent, determined Content : 98.0 per cent to 101.5 per cent (dried substance).
on 1.000 g by drying in an oven at 105 °C.
CHARACTERS
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g. Appearance : white or almost white, crystalline powder.
Solubility : freely soluble in water, sparingly soluble in ethanol
ASSAY (96 per cent), practically insoluble in methylene chloride.
Dissolve 0.250 g in water R and dilute to 100.0 mL with the
same solvent. To 20.0 mL of the solution add 100 mL of IDENTIFICATION
water R, 20 mL of hydrochloric acid R, 0.2 g of potassium First identification : B, C, E.
bromide R and 0.05 mL of methyl red solution R. Titrate Second identification : A, C, D, E.
dropwise with 0.0167 M potassium bromate, shaking A. Melting point (2.2.14) : 165 °C to 170 °C, with
continuously, until the red colour disappears. decomposition.
1 mL of 0.0167 M potassium bromate is equivalent to 3.429 mg B. Infrared absorption spectrophotometry (2.2.24).
of C6H7N3O. Comparison : isoprenaline hydrochloride CRS.
IMPURITIES C. Optical rotation (see Tests).
Specified impurities : A, B, E. D. To 0.1 mL of solution S (see Tests) add 0.05 mL of ferric
Other detectable impurities (the following substances would, chloride solution R1 and 0.9 mL of water R. A green colour
if present at a sufficient level, be detected by one or other of is produced. Add dropwise sodium hydrogen carbonate
the tests in the monograph. They are limited by the general solution R. The colour becomes blue and then red.
acceptance criterion for other/unspecified impurities and/or E. To 0.5 mL of solution S add 1.5 mL of water R. The solution
by the general monograph Substances for pharmaceutical use gives reaction (a) of chlorides (2.3.1).
(2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.10. Control of TESTS
impurities in substances for pharmaceutical use) : C, D. Prepare the solutions immediately before use.
Solution S. Dissolve 2.5 g in carbon dioxide-free water R and
dilute to 25.0 mL with the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than reference solution B7 or BY7
A. pyridine-4-carboxylic acid (isonicotinic acid), (2.2.2, Method II).
pH (2.2.3) : 4.3 to 5.5.
Mix 5 mL of solution S and 5 mL of carbon dioxide-free
water R.
Optical rotation (2.2.7) : − 0.10° to + 0.10°, determined on
B. pyridine-4-carboxamide (isonicotinamide), solution S.

3140 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 11.0 Isoprenaline sulfate

Related substances. Liquid chromatography (2.2.29).

Test solution. Dissolve 50.0 mg of the substance to be
examined in the mobile phase and dilute to 10.0 mL with the
mobile phase.
Reference solution (a). Dilute 1.0 mL of the test solution to
A. 1-(3,4-dihydroxyphenyl)-2-[(1-methylethyl)amino]-
100.0 mL with the mobile phase. Dilute 5.0 mL of this solution
ethanone.
to 10.0 mL with the mobile phase.
Reference solution (b). Dissolve 2.5 mg of orciprenaline
sulfate CRS in the mobile phase and dilute to 100 mL with 01/2008:0502
the mobile phase. corrected 10.0
Reference solution (c). To 5 mL of reference solution (a) add
5 mL of reference solution (b).
Reference solution (d). Dissolve 6.0 mg of isoprenaline
impurity A CRS in the mobile phase and dilute to 50.0 mL
with the mobile phase. Dilute 10.0 mL of the solution to ISOPRENALINE SULFATE
50.0 mL with the mobile phase.
Column :
Isoprenalini sulfas
– size : l = 0.125 m, Ø = 4.0 mm ;
– stationary phase : octadecylsilyl silica gel for
chromatography R (5 μm).
Mobile phase : methanol R, 11.5 g/L solution of phosphoric
acid R (5:95 V/V).
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 280 nm.
Injection : 20 μL. C22H36N2O10S,2H2O Mr 556.6
[6700-39-6]
Run time : 7 times the retention time of isoprenaline.
Identification of impurities : use the chromatogram obtained DEFINITION
with reference solution (d) to identify the peak due to Bis[(1RS)-1-(3,4-dihydroxyphenyl)-2-[(1-methylethyl)-
impurity A ; use the chromatogram obtained with reference amino]ethanol] sulfate dihydrate.
solution (b) to identify the peak due to orciprenaline. Content : 98.0 per cent to 102.0 per cent (anhydrous substance).
Relative retention with reference to isoprenaline
(retention time = about 4 min) : orciprenaline = about 1.5 ; CHARACTERS
impurity A = about 1.8. If necessary, adjust the concentration Appearance : white or almost white, crystalline powder.
of methanol in the mobile phase. Solubility : freely soluble in water, very slightly soluble in
System suitability : reference solution (c) : ethanol (96 per cent).
– resolution : minimum 3.0 between the peaks due to mp : about 128 °C, with decomposition.
isoprenaline and orciprenaline.
Limits : IDENTIFICATION
– impurity A : not more than the area of the corresponding First identification : A, D.
peak in the chromatogram obtained with reference Second identification : B, C, D.
solution (d) (0.5 per cent) ; A. Infrared absorption spectrophotometry (2.2.24).
– unspecified impurities : for each impurity, not more than Dissolve 0.5 g in 1.5 mL of water R and add 3.5 mL of
0.2 times the area of the principal peak in the chromatogram 2-propanol R. Scratch the wall of the tube with a glass rod
obtained with reference solution (a) (0.10 per cent) ; to initiate crystallisation. Collect the crystals and dry in
– total : maximum 1.0 per cent ; vacuo (2.2.32) at 60 °C.
– disregard limit : 0.1 times the area of the principal peak in Comparison : repeat the operations using 0.5 g of
the chromatogram obtained with reference solution (a) isoprenaline sulfate CRS.
(0.05 per cent). B. To 0.1 mL of solution S (see Tests) add 0.9 mL of water R
Loss on drying (2.2.32) : maximum 1.0 per cent, determined and 0.05 mL of ferric chloride solution R1. A green colour
on 1.000 g by drying in vacuo at 15-25 °C for 4 h. is produced. Add dropwise sodium hydrogen carbonate
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on solution R. The colour becomes blue and then red.
1.0 g. C. Dilute 1 mL of solution S to 10 mL with water R and add
0.25 mL of silver nitrate solution R1. A shining, grey,
ASSAY fine precipitate is formed within 10 min and the solution
In order to avoid overheating in the reaction medium, mix becomes pink.
thoroughly throughout and stop the titration immediately after D. Solution S gives reaction (a) of sulfates (2.3.1).
the end-point has been reached.
Dissolve 0.150 g in 10 mL of anhydrous formic acid R and add TESTS
50 mL of acetic anhydride R. Titrate with 0.1 M perchloric Solution S. Dissolve 5.0 g in carbon dioxide-free water R and
acid, determining the end-point potentiometrically (2.2.20). dilute to 50 mL with the same solvent. Use within 2 h of
1 mL of 0.1 M perchloric acid is equivalent to 24.77 mg preparation.
of C11H18ClNO3. Appearance of solution. Solution S is clear (2.2.1) and not
STORAGE more intensely coloured than reference solution Y6 (2.2.2,
Method II).
In an airtight container, protected from light.
pH (2.2.3) : 4.3 to 5.5.
IMPURITIES Dilute 5 mL of solution S to 10 mL with carbon dioxide-free
Specified impurities : A. water R.

General Notices (1) apply to all monographs and other texts 3141