Accessed from 10.6.1.

1 by merck1 on Mon May 11 20:35:23 EDT 2015

USP 38 Official Monographs / Acarbose 1999

. CU = concentration of the Sample solution (mg/mL)

Acarbose Acceptance criteria: 95.0%–102.0% on the anhydrous
basis
IMPURITIES
• RESIDUE ON IGNITION 〈281〉
Sample: 1.0 g
Acceptance criteria: NMT 0.2%

Delete the following:

C25H43NO18 645.60
D-Glucose, O-4,6-dideoxy-4-[[[1S-(1α,4α,5β,6α)]-4,5,6-trihy- •• HEAVY METALS, Method II 〈231〉: NMT 20 ppm• (Official 1-
droxy-3-(hydroxymethyl)-2-cyclohexen-1-yl]amino]-α-D-
.

Dec-2015)
glucopyranosyl-(1→4)-O-α-D-glucopyranosyl-(1→4)-; • CHROMATOGRAPHIC PURITY
O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox- Mobile phase, System suitability solution, Sample so-
ymethyl)-2-cyclohexen-1-yl]amino}-α-D-glucopyranosyl- lution, and Chromatographic system: Proceed as di-
(1→4)-O-α-D-glucopyranosyl-(1→4)-D-glucose rected in the Assay.
[56180-94-0]. Diluted sample solution: Dilute 1.0 mL of the Sample
DEFINITION solution with water to 100.0 mL.
Acarbose is produced by certain strains of Actinoplanes Analysis
utahensis. It contains NLT 95.0% and NMT 102.0% of Samples: Sample solution and Diluted sample solution
acarbose (C25H43NO18), calculated on the anhydrous basis. Calculate the percentage of each impurity in the por-
tion of Acarbose taken:
IDENTIFICATION
• A. INFRARED ABSORPTION 〈197K〉 Result = (rU/rA) × (1/F) × 100
• B. The retention time of the major peak of the Sample
solution corresponds to that of the Standard solution, as rU = peak response of each impurity from the
obtained in the Assay. Sample solution
rA = peak response of the main acarbose peak from
ASSAY the Diluted sample solution
• PROCEDURE F = relative response factor for each impurity (see
Solution A: 0.6 mg/mL of monobasic potassium phos- Table 1)
phate and 0.35 mg/mL of dibasic sodium phosphate in Acceptance criteria: See Table 1.
water
Mobile phase: Acetonitrile and Solution A (3:1) Table 1
System suitability solution: 20 mg/mL of USP Acarbose
Relative Relative Acceptance
System Suitability Mixture RS in water

USP Monographs
Retention Response Criteria,
Standard solution: 20 mg/mL of USP Acarbose RS in
Name Time Factor NMT (%)
water
Sample solution: 20 mg/mL of Acarbose in water Impurity Aa . 0.9 1 0.6
Chromatographic system Impurity Bb . 0.8 1.6 0.5
(See Chromatography 〈621〉, System Suitability.) Impurity Cc . 1.2 1 1.5
Mode: LC Impurity Dd 0.5 1.33 1.0
Detector: UV 210 nm
.

Impurity Ee 1.7 0.8 0.2

Column: 4-mm × 25-cm; packing L8
.

Impurity Ff 1.9 0.8 0.3

Column temperature: 35° .

Flow rate: 2 mL/min Impurity Gg . 2.2 0.8 0.3

Injection volume: 10 µL Impurity Hh . 0.6 1 0.2
System suitability a O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
Sample: System suitability solution
.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
Identify the acarbose peak and the peaks due to the glucopyranosyl-(1→4)-D-arabino-hex-2-ulopyranose.
b (1R,4R,5S,6R)-4,5,6-Trihydroxy-2-(hydroxymethyl)cyclohex-2-enyl 4-O-
impurities listed in Table 1. .

Suitability requirements
Peak-to-valley ratio: The ratio of the height of the
impurity A peak to the height of the valley between
[4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-α-D-glucopyranoside.
c α-D-Glucopyranosyl 4-O-[4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-
.

3-(hydroxymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-α-D-
the impurity A peak and the acarbose peak is NLT glucopyranoside.
1.2. d 4-O-[4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

Chromatogram comparability: The chromatogram ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-D-glucopyranose.

e O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
obtained is similar to the chromatogram provided .

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
with USP Acarbose System Suitability Mixture RS for glucopyranosyl-(1→4)-O-α-D-glucopyranosyl-(1→4)-D-arabino-hex-2-
the known impurities found. ulopyranose (4-O-α-acarbosyl-D-fructopyranose).
Analysis f O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

Samples: Standard solution and Sample solution ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-

glucopyranosyl-(1→4)-O-α-D-glucopyranosyl-(1→4)-D-glucopyranose (4-O-
Calculate the percentage of acarbose (C25H43NO18) in α-acarbosyl-D-glucopyranose).
the portion of Acarbose taken: g α-D-Glucopyranosyl O-4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
.

Result = (rU/rS) × (CS/CU) × 100
rU = peak response from the Sample solution
(hydroxymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
glucopyranosyl-(1→4)-O-α-D-glucopyranoside (α-D-glucopyranosyl α-
acarboside).
h O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

rS = peak response from the Standard solution ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-6-deoxy-α-

D-glucopyranosyl-(1→4)-D-glucopyranose.
CS = concentration of USP Acarbose RS in the
Standard solution (mg/mL)

Official from May 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.
 Accessed from 10.6.1.1 by merck1 on Mon May 11 20:35:23 EDT 2015

2000 Acarbose / Official Monographs USP 38

Table 1 (Continued) Identification—

Relative Relative Acceptance A: Infrared Absorption 〈197K〉.
Retention Response Criteria, B: Prepare a mixture of the Standard preparation and the
Name Time Factor NMT (%) Assay preparation (1:1), and chromatograph the mixture as
Any individual directed in the Assay: the chromatogram thus obtained ex-
unknown im- — — hibits a single major peak due to acebutolol.
purity 0.2 C: It responds to the tests for Chloride 〈191〉, when tested
Total impuri- as directed for alkaloidal hydrochlorides.
— —
ties 3.0 pH 〈791〉: between 4.5 and 7.0, in a solution (1 in 100).
a O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

Melting range 〈741〉: between 140° and 144°.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
glucopyranosyl-(1→4)-D-arabino-hex-2-ulopyranose. Loss on drying 〈731〉—Dry it at 105° for 3 hours: it loses
b (1R,4R,5S,6R)-4,5,6-Trihydroxy-2-(hydroxymethyl)cyclohex-2-enyl 4-O-
.
not more than 1.0% of its weight.
[4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox- Residue on ignition 〈281〉: not more than 0.1%.
ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-α-D-glucopyranoside.
c α-D-Glucopyranosyl 4-O-[4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-
.

3-(hydroxymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-α-D-
glucopyranoside. Delete the following:
d 4-O-[4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl]-D-glucopyranose. •Heavy metals, Method II 〈231〉:

. 0.002%.• (Official 1-Dec-2015)
e O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
Chromatographic purity—
glucopyranosyl-(1→4)-O-α-D-glucopyranosyl-(1→4)-D-arabino-hex-2- Standard solution—Prepare a solution of USP Acebutolol
ulopyranose (4-O-α-acarbosyl-D-fructopyranose). Hydrochloride RS in methanol containing 1.0 mg per mL.
f O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D- Test solution 1—Prepare a solution of Acebutolol Hydro-

glucopyranosyl-(1→4)-O-α-D-glucopyranosyl-(1→4)-D-glucopyranose (4-O- chloride in methanol containing 10 mg per mL.
α-acarbosyl-D-glucopyranose). Test solution 2—Mix 1 mL of Test solution 1 and 9 mL of
g α-D-Glucopyranosyl O-4,6-dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
.

(hydroxymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-α-D-
methanol.
glucopyranosyl-(1→4)-O-α-D-glucopyranoside (α-D-glucopyranosyl α- Reference solution 1—Transfer 3.0 mL of the Standard so-
acarboside). lution to a 100-mL volumetric flask, dilute with methanol to
h O-4,6-Dideoxy-4-{[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydrox-
.

volume, and mix.

ymethyl)cyclohex-2-enyl]amino}-α-D-glucopyranosyl-(1→4)-O-6-deoxy-α-
D-glucopyranosyl-(1→4)-D-glucopyranose. Reference solution 2—Mix 5.0 mL of Reference solution 1
and 10.0 mL of methanol.
SPECIFIC TESTS Procedure—Apply separate 20-µL portions of the Standard
• OPTICAL ROTATION, Specific Rotation 〈781S〉 solution, Test solution 1, Test Solution 2, Reference solution 1,
Sample solution: 10 mg/mL in water and Reference solution 2 to a suitable thin-layer chromato-
Acceptance criteria: +168° to +183° graphic plate (see Thin-Layer Chromatography under Chro-
• PH 〈791〉 matography 〈621〉) coated with a 0.25-mm layer of chromat-
USP Monographs

Sample solution: 50 mg/mL ographic silica gel mixture. Allow the spots to dry, and
Acceptance criteria: 5.5–7.5 develop the chromatograms in a solvent system consisting
• WATER DETERMINATION, Method Ic 〈921〉: NMT 4.0% of the upper layer of a mixture of water, butyl alcohol, and
ADDITIONAL REQUIREMENTS glacial acetic acid (50:40:10) until the solvent front has
• PACKAGING AND STORAGE: Preserve in tight containers. moved about three-fourths of the length of the plate. Re-
• USP REFERENCE STANDARDS 〈11〉 move the plate from the developing chamber, mark the sol-
USP Acarbose RS vent front, and allow the plate to air-dry. Examine the plate
USP Acarbose System Suitability Mixture RS under short-wavelength UV light: the chromatograms from
Test solution 2 and the Standard solution show principal
spots at about the same RF value. No secondary spot in the
chromatogram from Test solution 1, excluding the area at
the point of application, is more intense than the principal
spot obtained from Reference solution 1 (0.3%), and not
Acebutolol Hydrochloride
.

more than two secondary spots in the chromatogram from

C18H28N2O4 · HCl 372.89
Butanamide, N-[3-acetyl-4-[2-hydroxy-3-[(1-methyleth-
yl)amino]propoxy]phenyl]-, monohydrochloride, (±)-.
(±)-3′-Acetyl-4′-[2-hydroxy-3-(isopropylamino)propoxy]-
butyranilide monohydrochloride [34381-68-5].
» Acebutolol Hydrochloride contains not less than
98.0 percent and not more than 102.0 percent of
C18H28N2O4 · HCl, calculated on the dried basis.
Packaging and storage—Preserve in tight containers, and
store at controlled room temperature.
USP Reference standards 〈11〉—
USP Acebutolol Hydrochloride RS
Test solution 1 are more intense than the principal spot ob-
tained from Reference solution 2 (0.1%), and the total of all
impurities detected in the chromatogram of Test solution 1 is
not more than 0.5%.
Assay—
Mobile phase—Prepare a filtered and degassed mixture of
methanol, a 0.3% aqueous solution of sodium dodecyl sul-
fate, and glacial acetic acid (675:325:20). Make adjustments
if necessary to achieve a retention time for acebutolol of
between 4 minutes and 7 minutes (see System Suitability
under Chromatography 〈621〉).
Standard preparation—Quantitatively dissolve an accu-
rately weighed quantity of USP Acebutolol Hydrochloride RS
in water to obtain a solution having a known concentration
of about 0.14 mg per mL.
Assay preparation—Transfer about 35 mg of Acebutolol
Hydrochloride, accurately weighed, to a 250-mL volumetric
flask, dilute with water to volume, and mix.
Chromatographic system (see Chromatography 〈621〉)—The
liquid chromatograph is equipped with a 254-nm detector
and a 3.9-mm × 30-cm column that contains packing L1.
The flow rate is about 2 mL per minute. Chromatograph the

Official from May 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.