5/30/2019 USP Monographs: Pamabrom

Search USP29 Go

Pamabrom

C11H18BrN5O3 348.20

8-Bromo-3,7-dihydro-1,3-dimethyl-1-H-purine-2,6-dione compound with 2-amino-2-methyl-1-propanol (1:1).

» Pamabrom contains not less than 72.2 percent and not more than 76.6 percent of 8-bromotheophylline (C7H7BrN4O2), calculated on the
anhydrous basis; and not less than 24.6 percent and not more than 26.6 percent of 2-amino-2-methyl-1-propanol (C4H11NO), calculated
on the anhydrous basis.
Packaging and storage— Preserve in well-closed containers.

USP Reference standards 11 — USP 8-Bromotheophylline RS. USP Theophylline RS.

http://ftp.uspbpep.com/v29240/usp29nf24s0_m60250.html 1/3
5/30/2019 USP Monographs: Pamabrom

Identification— It responds to the Thin-layer Chromatographic Identification Test 201 , using a solvent system consisting of a mixture of xylene, methanol, and
glacial acetic acid (11:2:1) and a Standard solution and a Test solution prepared as directed below: the RF value of the principal spot, which appears as a dark spot
against a green background, from the Test solution corresponds to that obtained from the Standard solution.

Standard solution— Transfer an accurately weighed quantity of about 20 mg of USP 8-Bromotheophylline RS to a 100-mL volumetric flask, add 25 mL of water, 50 mL
of methanol, and a small amount of dilute ammonium hydroxide. Swirl the flask to effect solution. Dilute the contents of the flask with methanol to volume, and mix.

Test solution— Transfer an accurately weighed quantity of about 25 mg of Pamabrom to a 100-mL volumetric flask, add 25 mL of water, and swirl to dissolve. Dilute
the contents of the flask with methanol to volume, and mix.

Water, Method I 921 : not more than 3%.

Heavy metals, Method II 231 : 20 µg per g.

Limit of theophylline—

Diluting solution, Mobile phase, and Chromatographic system— Proceed as directed in the Assay for 8-bromotheophylline.

Standard solution— Dissolve an accurately weighed quantity of USP Theophylline RS in Diluting solution, add a few drops of ammonium hydroxide, sonicating if
necessary, to obtain a solution having a known concentration of about 1 mg of USP Theophylline RS per mL. Dilute a volume of this solution quantitatively, and
stepwise if necessary, with Diluting solution to obtain a solution having a known concentration of about 5 µg per mL.

Test solution— Transfer an accurately weighed quantity of about 200 mg of Pamabrom to a 200-mL volumetric flask. Add about 50 mL of Diluting solution, and
sonicate for 5 minutes. Cool to room temperature, dilute with Diluting solution to volume, and mix.

Procedure— Separately inject equal volumes (about 20 µL) of the Standard solution and the Test solution into the chromatograph, record the chromatograms, and
measure the responses for the major peaks. Calculate the percentage of theophylline in the portion of Pamabrom taken by the formula:
20(C / W)(rU / rS),

in which C is the concentration, in µg per mL, of USP Theophylline RS in the Standard solution, W is the weight, in mg, of Pamabrom taken, and rU and rS are the peak
responses of theophylline obtained from the Test solution and the Standard solution, respectively: not more than 0.5% is found.

Residual solvents 467 : meets the requirements.

Assay for 8-bromotheophylline—

Diluting solution— Prepare a mixture of water and methanol (70:30).

Mobile phase— Prepare a filtered and degassed mixture of water, methanol, and glacial acetic acid (69:30:1), filter, and degas. Make adjustments if necessary (see
System Suitability under Chromatography 621 ).

http://ftp.uspbpep.com/v29240/usp29nf24s0_m60250.html 2/3
5/30/2019 USP Monographs: Pamabrom

Internal standard solution— Dissolve an accurately weighed quantity of caffeine in Diluting solution, and dilute quantitatively, and stepwise if necessary, to obtain a
solution having a concentration of about 125 µg of caffeine per mL.

Standard preparation— Dissolve an accurately weighed quantity of USP 8-Bromotheophylline RS in Diluting solution, add a few drops of ammonium hydroxide,
sonicating if necessary, to obtain a solution having a known concentration of about 0.75 mg of USP 8-Bromotheophylline RS per mL. Transfer 5.0 mL of this solution to
a 100-mL volumetric flask, add 10.0 mL of Internal standard solution, dilute with Mobile phase to volume, mix, and filter.

Assay preparation— Transfer an accurately weighed quantity of about 200 mg of Pamabrom to a 200-mL volumetric flask, add about 50 mL of Diluting solution and
two drops of ammonium hydroxide, and sonicate for 5 minutes. [NOTE—If a hazy solution is present after 5 minutes of sonication, add 1 additional drop of ammonium
hydroxide.] Cool, dilute with Diluting solution to volume, and mix. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, add 10.0 mL of Internal standard
solution, dilute with Mobile phase to volume, mix, and filter.

Chromatographic system (see Chromatography 621 )—The liquid chromatograph is equipped with a 280-nm detector and a 4.6-mm × 15-cm column that contains
packing L1. The flow rate is about 1.5 mL per minute. Chromatograph 20 µL of the Standard preparation, and record the peak responses as directed under Procedure:
the relative retention times are about 0.6 for caffeine and 1.0 for 8-bromotheophylline, the resolution, R, between caffeine and 8-bromotheophylline 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 responses for the major peaks. Calculate the quantity, in mg, of 8-bromotheophylline (C7H7BrN4O2) in the portion of Pamabrom
taken by the formula:
4000C(RU / RS),

in which C is the concentration, in mg per mL, of USP 8-Bromotheophylline RS in the Standard preparation, and RU and RS are the peak response ratios of the 8-
bromotheophylline peak and the internal standard peak obtained from the Assay preparation and the Standard preparation, respectively.

Assay for 2-amino-2-methyl-1-propanol— Dissolve about 1 g of Pamabrom, accurately weighed, in 10 mL of water by warming gently on a steam bath until the
solution is clear. Cool, add methyl orange TS, and titrate with 0.5 N hydrochloric acid VS. Perform a blank determination, and make any necessary correction (see
Titrimetry 541 ). Each mL of 0.5 N hydrochloric acid is equivalent to 44.57 mg of C4H11NO.

Auxiliary Information— Staff Liaison : Andrzej Wilk, Ph.D., Senior Scientific Associate
Expert Committee : (MDCV05) Monograph Development-Cardiovascular
USP29–NF24 Page 1628
Pharmacopeial Forum : Volume No. 26(6) Page 1562
Phone Number : 1-301-816-8305

http://ftp.uspbpep.com/v29240/usp29nf24s0_m60250.html 3/3