IMPURITY PROFILING IMPURITY PROFILING OF THIAMINE HYDROCHLORIDE INJECTION BY RP-HPLC AND CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF
CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF
Objective: To propose comprehensive, simple and affordable RP-HPLC method for impurity profiling and characterization of unknown degradation product of Thiamine hydrochloride injectable formulation.
The chromatographic separation employs gradient mode using octadecyl silane column using mobile phase consist of Phosphate buffer with ion pair reagent, acetonitrile and methanol delivered flow rate at 1.2 ml/min. The detection was carried out at 248 nm using empower software. LC-MS/MS/QTOF hyphenated technique used for isolation and characterization of unknown degradation impurity. The performance of the method was systematically validated as per ICH Q2 (R2) guidelines.
Degradation product observed in accelerated stability was characterized by LC-MS/MS/QTOF hyphenated technique and found m/z value 351.1604 and postulated as oxidative degradation product of Thiamine due to excipient interaction. The validated method was sensitive, selective and specific data proves the method is precise and accurate from LOQ to 150% level and results are within 95-108% and less than 4.5% RSD. Method is linear from 0.03-58.83 µg/ml with correlation coefficient more than 0.990 and LOD and LOQ value ranged from 0.03 to 1.51 μg/ml.
A simple, selective and efficient RP-HPLC method for impurity profiling of Thiamine injectable formulation was successfully developed and validated. Moreover, degradant unknown product observed above identification threshold was identified and characterized by LC-MS/MS/QTOF hyphenated techniques. The method is systematically validated and found to be specific, precise, Linear, accurate, stable, rugged and robust and promises to be comprehensive simple affordable technique for impurity profiling in Thiamine hydrochloride injectable formulation in quality control department.
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