Alprazolam
(al pra' zoe lam).
Click to View Image

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%
•  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.
ADDITIONAL REQUIREMENTS
•  Packaging and Storage: Preserve in tight containers, and store at controlled room temperature.
•  USP Reference Standards 11
USP Alprazolam RS Click to View Structure
USP Alprazolam Related Compound A RS Click to View Structure
2-(2-Acetylhydrazino)-7-chloro-5-phenyl-3H-1,4-benzodiazepine.
USP 2-Amino-5-chlorobenzophenone RS Click to View Structure
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