Packaging and storage. Preserve in single dose containers, preferably of Type I glass. 

Reference Standards: IQB-9302.HCl Reference Standard is supplied by the manufacturer. USP Endotoxin RS. Bupivacaine Hydrochloride RS. 

Identification:  
A: Dilute a volume of Injection equivalent to about 50 mg of IQB-9302 hydrochloride, with 0.01 N hydrochloric acid to 25 mL and proceed as directed under Identification of Organic Nitrogenous Bases. The injection meets the requirements of the test. 

B: The retention time of the IQB-9302.HCl peak in the chromatogram of the Assay preparation corresponds to that of IQB-9302.HCl peak in the chromatogram of standard preparation obtained as directed in the Assay. 

Bacterial endotoxins: It contains not more than 2.5 USP Endotoxin Units per mg of IQB-9302 hydrochloride. 

pH: between 4.6 and 5.4. 

Sterility: must be sterile. 

Particulate matter: not more than 6000 particles > 10 mm and not more than 600 > 25 mm/ampoule. 

Assay 

pH 6.8 phosphate buffer- Dissolve 1.94 g of monobasic potassium phosphate and 2.48 g of dibasic potassium phosphate in 1000 mL of water. Adjust, if necessary, with 1 N potassium  hydroxide or 1 N phosphoric acid to a pH of 6.8.  

Mobile phase-Prepare a fresh solution of acetonitrile and pH 6.8 phosphate buffer (60:40).  Adjust, if necessary, with 1 N phosphoric acid to a pH of 7.7 ± 0.2. Filter the solution through a membrane filter of 1 mm or finer porosity, and degas.  

Internal standard solution-Prepare a solution of bupivacaine hydrochloride in the mobile phase,  containing about 1.0 mg por mL.  

Standard preparation- Dissolve about 50 mg of IQB-9302 Hydrochioride RS, accurately  
weighed, in 10.0 mL of water, using sonication if necessary, in a 100-mL volumetric flask. Add 10 mL of Internal standard solution, dilute mobile phase to volume, and mix.  

Assay preparation- Transfer an accurately measured volume of Injection, equivalent to about 50  mg of IQB-9302  hydrochloride, to a 100-mL volumetric flask, add 10.0 mL of Internal standard solution, dilute with mobile phase to volume, and mix.  

Chromatographic system- The liquid chromatograph is equipped with a 262-nm detector and a 4-mm X 30-cm column that contains packing LI. The flow rate is about 1,6 mL por minute.  
Chromatograph three replicate injections of the Standard preparation, and record the peak  responses as directed under Procedure: the relative standard deviation of the ratios of the  bupivacaine hydrochloride peak to the IQB-9302 hydrochloride peak is not more than 1.0%, and the resolution factor between bupivacaine hydrochloride and IQB-9302 hydrochloride is not less than 2.0.  

Procedure- Separately inject equal volumes (about 20 mL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. The relative retention times are about 2.9 for IQB-9302 hydrochloride and 2.1 for bupivacaine hydrochloride. Calculate the quantity, in mg,  of C₁₈H₂₆N₂O.HCl in the volume of Injection taken by the formula:  

                     W(RU/RS),  

in which W is the weight, in mg, of IQB-9302 Hydrochloride RS, calculated on the  
 anhydrous basis, in the Standard preparation, and RU and RS are the ratios of the peak responses of IQB-9302 hydrochloride to those of the internal standard obtained from the Assay preparation and the Standard preparation, respectively.