DEVELOPMENT AND VALIDATION OF A HPLC METHOD FOR DETERMINATION OF RESIDUAL ETHYLENEDIAMINE IN DRUG SUBSTANCES

摘要

A HPLC method for determination of residual ethylenediamine in drug substances was developed and subsequently validated as per guidelines from ICJH, USP and other regulatory agencies. UV activity in ethylenediamine was induced by pre-column derivatization with 1-naphthyl isothiocyanate. Analysis was performed by using a C-18 HPLC column, gradient elution with pH 3 ortho-phosphoric acid and triethylamine buffer and methanol as mobile phase. UV 220 nm was found as suitable pi_(max)for detection. The method was validated for its specificity, precision, accuracy, linearity, ruggedness and robustness. Correlation coefficient for ethylenediamine peak area was found to be 0.99963 for linearity ranging between 0.025 mcg/mL to 0.750 mcg/mL. Limit of quantitation of the method was 0.025 mcg/mL and limit of detection was 0.015 mcg/mL.