Alprazolam
C17H13ClN4 308.76
4H-[1,2,4]Triazolo[4,3-
][1,4]benzodiazepine, 8-chloro-1-methyl-6-phenyl-;
8-Chloro-1-methyl-6-phenyl-4H-s-triazolo[4,3-][1,4]benzodiazepine
[28981-97-7].
(al pra' zoe lam).
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C17H13ClN4 308.76
4H-[1,2,4]Triazolo[4,3-
][1,4]benzodiazepine, 8-chloro-1-methyl-6-phenyl-;
8-Chloro-1-methyl-6-phenyl-4H-s-triazolo[4,3-
][1,4]benzodiazepine
[28981-97-7].DEFINITION
Alprazolam contains NLT 98.0% and NMT 102.0% of C17H13ClN4.
[Caution—Care should be taken to prevent inhaling particles of Alprazolam and exposing the skin to it.
]
IDENTIFICATION
• B.
The retention time of the major peak from the Sample solution corresponds to that from the Standard solution, as obtained in the Assay.
ASSAY
• Procedure
Diluent:
Acetonitrile and water (1:1)
Buffer:
1.4 g/L of monobasic potassium phosphate in water
Mobile phase:
Acetonitrile and Buffer (1:1)
Standard solution:
25 µg/mL of USP Alprazolam RS in Diluent. [Note—The solution is stable for 48 h at room temperature when stored in closed containers. ]
Sample solution:
25 µg/mL of Alprazolam in Diluent. Sonicate for about 1 min. [Note—The solution is stable for 48 h at room temperature when stored in closed containers. ]
Chromatographic system
Mode:
LC
Detector:
UV 231 nm
Column:
4.6-mm × 25-cm; packing L1
Column temperature:
40
Flow rate:
1 mL/min
Injection size:
20 µL
System suitability
Sample:
Standard solution
Suitability requirements
Tailing factor:
NMT 2.0
Relative standard deviation:
NMT 2.0%
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of alprazolam (C17H13ClN4) in the portion of Alprazolam taken:
Result = (rU/rS) × (CS/CU) × 100
| rU | = | = peak area from the Sample solution |
| rS | = | = peak area from the Standard solution |
| CS | = | = concentration of USP Alprazolam RS in the Standard solution (mg/mL) |
| CU | = | = concentration of Alprazolam in the Sample solution (mg/mL) |
Acceptance criteria:
98.0%–102.0%
IMPURITIES
• Residue on Ignition 
281
:
NMT 0.5%
281:
NMT 0.5%
• Heavy Metals, Method II 231:
20 ppm
231:
20 ppm
• Organic Impurities
Diluent, Buffer, Mobile phase, and Chromatographic system:
Proceed as directed in the Assay.
System suitability solution:
20 µg/mL each of USP Alprazolam RS, USP Alprazolam Related Compound A RS, and USP 2-Amino-5-chlorobenzophenone RS in Diluent
Standard solution:
0.25 µg/mL of USP Alprazolam RS in Diluent. [Note—When stored in closed containers, the solution is stable for 48 h at room temperature. ]
Sample solution:
250 µg/mL of Alprazolam in Diluent. Sonicate for about 1 min. [Note—When stored in closed containers, the Sample solution is stable for 24 h at room temperature. ]
System suitability
Samples:
Standard solution and System suitability solution
[Note—For relative retention times, see Table 1. ]
Suitability requirements
Resolution:
NLT 2.0 between alprazolam related compound A and alprazolam, System suitability solution
Relative standard deviation:
NMT 5.0%, Standard solution
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Alprazolam taken:
Result = (rU/rS) × (CS/CU) × (1/F) × 100
| rU | = | = peak response for each impurity in the Sample solution |
| rS | = | = peak response for alprazolam from the Standard solution |
| CS | = | = concentration of USP Alprazolam RS in the Standard solution (µg/mL) |
| CU | = | = concentration of Alprazolam in the Sample solution (µg/mL) |
| F | = | = relative response factor (see Table 1) |
Acceptance criteria:
See Table 1.
Table 1
| Name | Relative Retention Time |
Relative Response Factor |
Acceptance Criteria, NMT (%) |
|---|---|---|---|
| Alprazolam related compound A | 0.8 | 0.76 | 0.15 |
| Alprazolam | 1.0 | 1.0 | — |
| 2-Amino-5-chlorobenzophenone | 4.0 | 1.0 | 0.15 |
| Individual unspecified impurity | — | 1.0 | 0.10 |
| Total impurities | — | — | 1.0 |
SPECIFIC TESTS
• Loss on Drying 731:
Dry a sample at 105 for 1 h: it loses NMT 0.5% of its weight.
731:
Dry a sample at 105 for 1 h: it loses NMT 0.5% of its weight.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers, and store at controlled room temperature.
• USP Reference Standards 11
11
USP Alprazolam RS 
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USP Alprazolam Related Compound A RS
2-(2-Acetylhydrazino)-7-chloro-5-phenyl-3H-1,4-benzodiazepine.
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2-(2-Acetylhydrazino)-7-chloro-5-phenyl-3H-1,4-benzodiazepine.
USP 2-Amino-5-chlorobenzophenone RS
2-Amino-5-chlorobenzophenone.
C13H10ClNO 231.68
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2-Amino-5-chlorobenzophenone.
C13H10ClNO 231.68
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Ravi Ravichandran, Ph.D.
Principal Scientific Liaison 1-301-816-8330 |
(SM42010) Monographs - Small Molecules 4 |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 2102
Pharmacopeial Forum: Volume No. 33(5) Page 868