Objective:To establish a novel High Performance Liquid Chromatography(HPLC)method for the determining pipecolic acids and their derivatives(PADs)in drug substance.Methods:2,4-dinitrofluorobenzene(DNFB)was used as a derivative reagent,chromatographic column with octadecylsilane bonded silica gel as the filler was used.And water containing 0.1%phosphoric acid-ace-tonitrile system was utilized as a mobile phase with UV detector,and external standard method was used to quantity.Results:All PADs in different solvents had good linear relationship in between the range of the limit of quantitation(LOQ)to 300%with respect to specifica-tion limit,and all the linear correlation coefficients(r)were greater than 0.999.The spiked recoveries of LOQ were all between 85%and 110%,and relative standard deviation(RSDs)were all less than 4%.The spiked recoveries of 50%to 200%with respect to specification limit were all between 90%and 108%,and RSDs were all less than 3%.The limit of detection(LODs)were all less than 0.03 μg/mL(0.007 5%with respect to the concentration of the sample solution),LOQs were all less than 0.08 μg/mL(0.020%with respect to the concentration of the sample solution).Conclusion:This HPLC method has high sensitivity,strong universality,and high accuracy,which can be used as a universal method for quantitative determining PADs in drug substance.
维普
万方数据