Packaging and storage—
Preserve in tight, light-resistant containers, and store at controlled room temperature.
Identification—
Dissolve an amount of finely po
wdered 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 alprazolam–potassium 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 cm
1; at 932, 891, 826, 779, 746, 696, and 658 wavenumbers in the region of 975 to 600 cm
1.
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 under
Alprazolam.
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 C
17H
13ClN
4 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.
Assay—
Mobile phase, Chromatographic system, Internal standard solution, and Standard preparation—
Prepare as directed in the
Assay under
Alprazolam.
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.
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 (C
17H
13ClN
4) 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—
Staff Liaison :
Ravi Ravichandran, Ph.D., Senior Scientist
Expert Committee : (MDPP05) Monograph Development-Psychiatrics and Psychoactives
USP29–NF24 Page 78
Pharmacopeial Forum : Volume No. 30(5) Page 1582
Phone Number : 1-301-816-8330
