» Alprazolam Tablets contain not less than 90.0 percent and not more than 110.0 percent of the labeled amount of alprazolam (C17H13ClN4).
Packaging and storage Preserve in tight, light-resistant containers, and store at controlled room temperature.
Identification Dissolve an amount of finely powdered Tablets, equivalent to about 15 mg of alprazolam, in 10 mL of sodium carbonate solution (1 in 100). Add 15 mL of chloroform, and shake vigorously for 30 minutes. Centrifuge, withdraw the aqueous layer, and transfer the chloroform to a clean container. Add about 200 mg of potassium bromide. Evaporate the chloroform from this mixture to dryness, and dry the dispersion in vacuum at 60 for 24 hours. Grind this dispersion into a fine powder. Prepare a suitable pellet for testing by placing about 100 mg of dried potassium bromide into a die. Sprinkle about 20 mg of the finely ground alprazolampotassium bromide dispersion onto the dried potassium bromide layer, and cover with another specimen of about 100 mg of dried potassium bromide: the IR absorption spectrum of the potassium bromide dispersion so obtained exhibits maxima characteristic of alprazolam, as compared to that of a similar preparation of USP Alprazolam RS, at the following wavenumbers: at 1609, 1578, 1566, 1539, 1487, and 1379 wavenumbers in the region of 1650 to 1300 cm1; at 932, 891, 826, 779, 746, 696, and 658 wavenumbers in the region of 975 to 600 cm1.
Dissolution, Procedure for a Pooled Sample 711
Buffer stock solution Dissolve 160 g of monobasic potassium phosphate and 40 g of dibasic potassium phosphate in water, and dilute with water to obtain 2.0 L of solution. Add, with mixing, phosphoric acid or potassium hydroxide solution (45 in 100), as necessary to adjust the solution such that, when this Buffer stock solution is diluted 1 in 10 with water, the resulting solution has a pH of 6.0 ± 0.1.
Buffer solution Prepare a 1 in 10 dilution of Buffer stock solution in water to obtain a Working buffer solution having a pH of 6.0 ± 0.1.
Medium: Buffer solution; 500 mL.
Apparatus 1: 100 rpm.
Time: 30 minutes.
Determine the amount of C17H13ClN4 dissolved employing the following method.
Mobile phase Prepare a degassed and filtered solution of Buffer solution, acetonitrile, and tetrahydrofuran (60:35:5). Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard stock solution Prepare a solution in methanol of USP Alprazolam RS having a known concentration of about 0.05 mg per mL.
Standard solution Add 50 mL of Buffer stock solution and 250 mL of water to a 500-mL volumetric flask. Add to the flask 5.0 mL of Standard stock solution for every 0.25 mg of alprazolam contained in the Tablet being assayed. Dilute with water to volume, and mix.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 254-nm detector and a 4.6-mm × 10-cm analytical column that contains packing L7. The flow rate is about 1 mL per minute. Chromatograph the Standard solution, and record the peak responses as directed for Procedure: the column efficiency is not less than 500 theoretical plates, and the relative standard deviation for replicate injections is not more than 3.0%.
Procedure Separately inject equal volumes of a filtered portion of the solution under test and the Standard solution into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the quantity of C17H13ClN4 dissolved based on the peak responses obtained from the solution under test and the Standard solution.
Tolerances Not less than 80% (Q) of the labeled amount of C17H13ClN4 is dissolved in 30 minutes.
Uniformity of dosage units 905: meet the requirements for Content Uniformity.
procedure for content uniformity
Mobile phase Prepare as directed in the Assay.
Internal standard solution Prepare a solution of triazolam in acetonitrile having a concentration of about 0.032 mg per mL.
Test preparation Transfer 1 Tablet to a container. Add about 0.4 mL of water directly onto the Tablet, allow the Tablet to stand for 2 minutes, and then swirl the container to disperse the Tablet. For every 0.25 mg of alprazolam contained in the Tablet, add 10.0 mL of Internal standard solution to the container. Shake, and centrifuge if necessary.
Standard preparation Prepare a solution in Internal standard solution of USP Alprazolam RS having a known concentration of about 0.025 mg per mL.
Chromatographic system and Procedure Proceed as directed in the Assay under Alprazolam. Calculate the quantity, in mg, of C17H13ClN4 in the Tablet by the formula:
CV(RU / RS)where V is the volume, in mL, of Internal standard solution in the Test preparation; and the other terms are as defined in the Procedure for Alprazolam.
Mobile phase Prepare a filtered and degassed mixture of acetonitrile, chloroform, butyl alcohol, water, and glacial acetic acid (850:80:50:20:0.5). Make adjustments if necessary (see System Suitability under Chromatography 621).
Internal standard solution Dissolve an accurately weighed quantity of triazolam in acetonitrile, and dilute with acetonitrile to obtain a solution having a known concentration of about 0.25 mg per mL.
Standard preparation Dissolve an accurately weighed quantity of USP Alprazolam RS in Internal standard solution, and dilute with Internal standard solution to obtain a solution having a known concentration of about 0.25 mg per mL. Transfer 5.0 mL of this solution to a 50-mL volumetric flask, dilute with acetonitrile to volume, and mix.
Assay preparation Weigh and finely powder not fewer than 20 Tablets. Transfer an accurately weighed quantity of the powder, equivalent to about 5 mg of alprazolam, to a 200-mL volumetric flask. Transfer 2 mL of water and 20 mL of Internal standard solution, shake vigorously for 10 minutes, dilute with acetonitrile to volume, and mix.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 254-nm detector and a 4.6-mm × 30-cm column that contains packing L3. The flow rate is about 2 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the resolution, R, between the internal standard and alprazolam is not less than 2.0; and the relative standard deviation for replicate injections 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, record the chromatograms, and measure the area responses for the major peaks. Calculate the quantity, in mg, of alprazolam (C17H13ClN4) in the portion of Tablets taken by the formula:
200C(RU / RS)in which C is the concentration, in mg per mL, of USP Alprazolam RS in the Standard preparation; and RU and RS are the peak response ratios of the alprazolam peak relative to the internal standard peak 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 1455Pharmacopeial Forum: Volume No. 33(1) Page 41
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.