Printed on: Tue May 23 2023, 02:12:31 AM(EST) Status: Currently Official on 23-May-2023 DocId: GUID-195CFE4C-C2A0-4921-B42F-BC390A45A710_4_en-US

Printed by: Anandkumar Joshi Official Date: Official as of 01-May-2020 Document Type: USP @2023 USPC
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1

responses for the major peaks. Calculate the percentage of

Diazepam diazepam related compound B, diazepam related compound
A, and nordazepam in the portion of Diazepam taken by the
formula:

(CR/W)(rU/rS)

in which CR is the concentration, in µg per mL, of USP

Diazepam Related Compound B RS, USP Diazepam Related
Compound A RS, or USP Nordazepam RS in the Standard
C16H13ClN2O 284.74 solution; W is the weight, in mg, of Diazepam taken to prepare
2H-1,4-Benzodiazepin-2-one, 7-chloro-1,3-dihydro-1- the Test solution; and rU and rS are the peak responses obtained
methyl-5-phenyl-. from the Test solution and the Standard solution, respectively:
7-Chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4- not more than 0.01% of diazepam related compound A, not
benzodiazepin-2-one CAS RN®: 439-14-5; UNII: more than 0.1% of diazepam related compound B, and not
Q3JTX2Q7TU. more than 0.3% of nordazepam are found.
Calculate the percentage of any other impurity in the portion
» Diazepam contains not less than 95.0 percent of Diazepam taken by the formula:
and not more than 105.0 percent of C16H13ClN2O, (CS/W)(ri/rS)
calculated on the dried basis.
in which CS is the concentration, in µg per mL, of USP
Packaging and storage—Preserve in tight, light-resistant

al
Diazepam Related Compound B RS in the Standard solution; ri
containers. is the peak response for any other impurity obtained from the
USP Reference standards á11ñ— Test solution; and rS is the peak response of USP Diazepam
USP Diazepam RS Related Compound B RS obtained from the Standard solution:
USP Diazepam Related Compound A RS not more than 0.1% of any other impurity is found; and not
2-Methylamino-5-chlorobenzophenone. more than 1.0% of the total impurities is found.
C14H12ClNO 245.71
USP Diazepam Related Compound B RS
3-Amino-6-chloro-1-methyl-4-phenylcarbostyril.
ci Assay—
Mobile phase—Prepare a filtered and degassed mixture of
C16H13ClN2O 284.74 acetonitrile, water, and methanol (2:2:1). Make adjustments
USP Nordazepam RS if necessary (see System Suitability under Chromatography
ffi
7-Chloro-1,3-dihydro-5-phenyl-2H-1,4-benzodiazepin- á621ñ).
2-one. System suitability solution—Dissolve suitable quantities of
C15H11ClN2O 270.72 USP Nordazepam RS and USP Diazepam RS in methanol, using
sonication if necessary, to obtain a solution containing about
Identification— 0.1 mg of each per mL.
Standard preparation—Dissolve an accurately weighed
Change to read:
O

quantity of USP Diazepam RS in methanol, and dilute

A: ▲Spectroscopic Identification Tests á197ñ, Infrared quantitatively, and stepwise if necessary, with methanol to
Spectroscopy: 197K▲ (CN 1-May-2020). obtain a solution having a known concentration of about
B: Thin-Layer Chromatographic Identification Test á201ñ— 0.1 mg per mL.
Test solution: 5 mg per mL, in acetone. Assay preparation—Transfer about 10 mg of Diazepam,
Developing solvent system: a mixture of ethyl acetate and accurately weighed, to a 100-mL volumetric flask, dissolve in
n-heptane (1:1). and dilute with methanol to volume, and mix.
Procedure—Proceed as directed in the chapter except use Chromatographic system (see Chromatography á621ñ)—
an unsaturated developing chamber. The liquid chromatograph is equipped with a 254-nm
Melting range, Class I á741ñ: between 131° and 135°. detector and a 3.9-mm × 15-cm column that contains packing
Loss on drying á731ñ—Dry it in vacuum over phosphorus L1. The flow rate is about 1 mL per minute. Chromatograph
pentoxide at 60° for 4 hours: it loses not more than 0.5% of the System suitability solution, and record the peak responses
its weight. as directed for Procedure: the relative retention times are about
Residue on ignition á281ñ: not more than 0.1%. 0.76 for nordazepam and 1.0 for diazepam; the resolution, R,
between nordazepam and diazepam is not less than 4; the
Related compounds— column efficiency is not less than 5000 theoretical plates for
Mobile phase, System suitability solution, and the diazepam peak; the tailing factor for diazepam is not more
Chromatographic system—Proceed as directed in the Assay. than 2.0; and the relative standard deviation for the diazepam
Standard solution—Dissolve accurately weighed quantities peak for replicate injections is not more than 2.0%.
of USP Diazepam Related Compound B RS, USP Diazepam Procedure—Separately inject equal volumes (about 10 µL)
Related Compound A RS, and USP Nordazepam RS in of the Standard preparation and the Assay preparation into the
methanol, and dilute quantitatively, and stepwise if necessary, chromatograph, record the chromatograms, and measure the
with methanol to obtain a solution having known responses for the major peaks. Calculate the quantity, in mg,
concentrations of about 1 µg per mL, 0.1 µg per mL, and 3 µg of C16H13ClN2O in the portion of Diazepam taken by the
per mL, respectively. formula:
Test solution—Transfer about 10 mg of Diazepam,
accurately weighed, to a 10-mL volumetric flask, dissolve in 100C(rU/rS)
and dilute with methanol to volume, and mix.
Procedure—Separately inject equal volumes (about 10 µL) in which C is the concentration, in mg per mL, of USP
of the Standard solution and the Test solution into the Diazepam RS in the Standard preparation; and rU and rS are the
chromatograph, record the chromatograms, and measure the

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Printed on: Tue May 23 2023, 02:12:31 AM(EST) Status: Currently Official on 23-May-2023 DocId: GUID-195CFE4C-C2A0-4921-B42F-BC390A45A710_4_en-US
Printed by: Anandkumar Joshi Official Date: Official as of 01-May-2020 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: gz3ku DOI: https://doi.org/10.31003/USPNF_M24640_04_01
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peak responses obtained from the Assay preparation and the

Standard preparation, respectively.

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