Atracurium Besylate Injection
» Atracurium Besylate Injection is a sterile solution containing not less than 90.0 percent and not more than 115.0 percent of the labeled amount of atracurium besylate (C65H82 N2O18S2). It contains an amount of the trans-trans-isomer equivalent to not less than 5.0 percent and not more than 6.5 percent of the labeled amount of atracurium besylate, an amount of the cis-trans-isomer equivalent to not less than 34.5 percent and not more than 38.5 percent of the labeled amount of atracurium besylate, and an amount of the cis-cis-isomer equivalent to not less than 55.0 percent and not more than 60.0 percent of the labeled amount of atracurium besylate.
note—The Injection is unstable at room temperature. Store all samples in the refrigerator. Analyze all preparations as soon as possible, or use a refrigerated injector.
Packaging and storage— Preserve in single-dose or multiple-dose containers, preferably of Type I glass, in a refrigerator, and protect from freezing. Protect from light.
Identification— The retention times of the peaks of the three atracurium besylate isomers in the chromatogram of the Assay preparation correspond to those in the chromatogram of the Standard preparation, as obtained in the Assay.
Bacterial endotoxins 85 It contains not more than 5.56 USP Endotoxin Units per mg of atracurium besylate.
Sterility 71 It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
pH 791: between 3.00 and 3.65.
Related compounds—
Buffer solution, Solution A, Solution B, Mobile phase, and Standard preparation— Proceed as directed in the Assay under Atracurium Besylate.
System suitability solution— Heat a portion of the Standard preparation at 90 for 30 minutes, and immediately chill to about 5.
Diluted standard preparation— Dilute a portion of the Standard preparation quantitatively, and stepwise if necessary, with Solution A to obtain a solution having a known concentration of about 0.02 mg per mL.
Test preparation— Use the Assay preparation.
Chromatographic system— Prepare as directed for Chromatographic system in the Assay. Chromatograph the System suitability solution and the Diluted standard preparation, record the chromatograms, and measure the responses for the degradation products by comparing the peak responses of the System suitability solution to those of the Diluted standard preparation as directed for Procedure: the retention times relative to the atracurium besylate cis-cis-isomer are about 0.22 for the acidic compound; 0.29 for laudanosine; 0.44 and 0.50 for the trans- and cis-isomers, respectively, of the hydroxy compound; and about 1.28 and 1.33 for the trans- and cis-isomers, respectively, of the monoacrylate.
Procedure— Separately inject equal volumes (about 20 µL) of the Diluted standard preparation and the Test preparation into the chromatograph, record the chromatograms, and measure the peak responses, except the peak due to benzenesulfonic acid occurring at a retention time of about 0.08 relative to the atracurium besylate cis-cis-isomer. Calculate the percentage of each impurity in the portion of Test preparation taken by the formula:
100(C/M)(ri / rs)
in which C is the concentration, in mg per mL, of USP Atracurium Besylate RS in the Diluted standard preparation; M is the concentration of atracurium besylate, in mg per mL, in the Test preparation; ri is the peak response for each impurity obtained from the Test preparation; and rs is the sum of the responses of all the peaks obtained from the Diluted standard preparation: not more than 6.0% of the acidic compound, not more than 6.0% of the combined cis- and trans-isomers of the hydroxy compound, not more than 3.0% of laudanosine, not more than 3.0% of the combined cis- and trans-isomers of the monoacrylate, and not more than 2.0% of other known synthetic impurities is found; not more than 0.1% of any other impurity is found; and not more than 15.0% of total impurities is found.
Other requirements— It meets the requirements under Injections 1.
Assay—
Buffer solution, Solution A, Solution B, Mobile phase, and Standard preparation— Proceed as directed in the Assay under Atracurium Besylate.
Assay preparation— Transfer an accurately measured volume of Injection, equivalent to about 50 mg of atracurium besylate, to a 50-mL volumetric flask, dilute with Solution A to volume, and mix.
Chromatographic system (see Chromatography 621)— The liquid chromatograph is equipped with a 280-nm detector and a 4.6-mm × 25-cm column that contains base-deactivated packing L1. The flow rate is about 1 mL per minute. The chromatograph is programmed as follows.
Time
(minutes)
Solution A
(%)
Solution B
(%)
Elution
0 80 20 equilibration
0–5 80 20 isocratic
5–15 80®40 20®60 linear gradient
15–25 40 60 isocratic
25–30 40®0 60®100 linear gradient
Chromatograph replicate injections of the Standard preparation, and record the peak responses as directed for Procedure: the relative retention times are about 0.8 for the atracurium besylate trans-trans-isomer, 0.9 for the cis-trans-isomer, and 1.0 for the cis-cis-isomer; the resolution, R, between the atracurium besylate trans-trans-isomer and the cis-trans-isomer and between the atracurium besylate cis-trans-isomer and the cis-cis-isomer 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 three atracurium besylate isomer peaks. Calculate the quantity, in mg, of atracurium besylate (C65H82N2O18S2) in each mL of the Injection taken by the formula:
50(C/V)(rU / rS)
in which C is the concentration, in mg per mL, of USP Atracurium Besylate RS in the Standard preparation; V is the volume, in mL, of Injection taken for the Assay preparation; and rU and rS are the sums of the peak responses of the atracurium besylate trans-trans, trans-cis, and cis-cis-isomers obtained from the Assay preparation and the Standard preparation, respectively.
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Topic/Question Contact Expert Committee
Monograph Ravi Ravichandran, Ph.D.
Senior Scientist
1-301-816-8330
(MDPP05) Monograph Development-Psychiatrics and Psychoactives
Reference Standards Lili Wang, Technical Services Scientist
1-301-816-8129
RSTech@usp.org
85 Radhakrishna S Tirumalai, Ph.D.
Senior Scientist
1-301-816-8339
(MSA05) Microbiology and Sterility Assurance
71 Radhakrishna S Tirumalai, Ph.D.
Senior Scientist
1-301-816-8339
(MSA05) Microbiology and Sterility Assurance
USP32–NF27 Page 1603
Pharmacopeial Forum: Volume No. 34(4) Page 905
Chromatographic Column—
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.