Alprazolam
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]. » Alprazolam contains not less than 98.0 percent and not more than 102.0 percent of C17H13ClN4.
CautionCare should be taken to prevent inhaling particles of Alprazolam and exposing the skin to it.
Packaging and storage
Preserve in tight containers, and store at controlled room temperature.
USP Reference standards 11
USP Alprazolam RS. USP Alprazolam Related Compound A RS. USP 2-Amino-5-chlorobenzophenone RS .
Identification
B:
The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
Loss on drying 731
Dry it at 105 for 1 hour: it loses not more than 0.5% of its weight.
Residue on ignition 281:
not more than 0.5%.
Heavy metals, Method II 231:
0.002%.
Related compounds
Diluent, Buffer, Mobile phase, and Standard preparation
Prepare as directed in Assay.
System suitability solution
Dissolve suitable quantities of USP Alprazolam RS, USP Alprazolam Related Compound A RS, and USP 2-Amino-5-chlorobenzophenone RS in Diluent to obtain a final concentration of 20 µg per mL of each.
Standard solution
Quantitatively dilute a suitable volume of the Standard preparation with Diluent to obtain a final solution having a known concentration of 0.25 µg per mL of alprazolam.
[noteWhen stored in closed containers, the Standard solution is stable for 48 hours at room temperature.]
Test solution
Use the Assay stock preparation.
Chromatographic system
Prepare as directed in Assay. Chromatograph separately the System suitability solution and the Standard solution, and record the peak responses as directed for Procedure: the resolution, R, between alprazolam related compound A and alprazolam is not less than 2.0; and the relative standard deviation for the alprazolam peak in the Standard solution is not more than 5.0%.
Procedure
Separately inject equal volumes (about 20 µL) of the Standard solution and the Test solution into the chromatograph, and record the chromatograms for up to a minimum of five times the retention time of alprazolam. Identify the peaks using the approximate relative retention times given in Table 1. Calculate the quantity, in percent of each of the related compounds and any of the unspecified impurities by the formula:
100(1/F)(CS / CT)(ri / rS)
in which, F is the relative response factor for each impurity (given in Table 1); CS and CT are the concentrations of alprazolam, in µg per mL, in the Standard solution and the Test solution, respectively; ri is the peak response for the related compound or unspecified impurity in the Test solution chromatogram; and rS is the peak response for alprazolam in the Standard solution chromatogram. The relative response factors and the limits of the related compounds are given in Table 1.
Table 1
Assay
Diluent
Prepare a mixture of acetonitrile and water (1:1).
Buffer
Dissolve 1.36 g of monobasic potassium phosphate (KH2PO4) in 1 L of water, and mix.
Mobile phase
Prepare a mixture of filtered and degassed acetonitrile and Buffer (1:1).
Standard preparation
Dissolve an accurately weighed quantity of USP Alprazolam RS in Diluent to obtain a solution having a known concentration of 25 µg per mL of alprazolam.
[noteWhen stored in closed containers, the Standard preparation is stable for 48 hours at room temperature.]
Assay stock preparation
Dissolve an accurately weighed quantity of Alprazolam in Diluent to obtain a solution having a known concentration of 250 µg per mL of alprazolam. Sonicate for about 1 minute. [noteWhen stored in closed containers, the Assay stock preparation is stable for 24 hours at room temperature.]
Assay preparation
Dilute a suitable volume of the Assay stock preparation with Diluent to obtain a solution having a known concentration of about 25 µg per mL of alprazolam
[noteWhen stored in closed containers, the Assay preparation is stable for 48 hours at room temperature.]
Chromatographic system
The liquid chromatograph is equipped with a 231-nm detector and a 4.6-mm × 25-cm column that contains packing L1. The flow rate is about 1 mL per minute. The column temperature is maintained at 40. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing factor is not more than 2.0 for the alprazolam peak, and the relative standard deviation of the alprazolam peak is not more than 2.0%.
Procedure
Separately inject equal volumes (about 20 µL) of the Standard preparation and the Assay preparation into the chromatograph, and measure the responses for the alprazolam peak. Calculate the quantity, in percent of C17H13ClN4 in the portion of Alprazolam taken by the formula:
100(CS / CU)(rU / rS)
in which, CS and CU are the concentrations of alprazolam, in µg per mL, in the Standard preparation and the Assay preparation, respectively; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information
Please check for your question in the FAQs before contacting USP.
Chromatographic Column
USP32NF27 Page 1454
Pharmacopeial Forum: Volume No. 33(5) Page 868
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.
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