Research on Identification and Determination of Multiple Components in Longjiatongluo Capsule

KeyWord:HPLC-MS/MS  multi-component determination  HPLC-DAD/ELSD  Longjiatongluo capsule

Abstract:

      An HPLC-MS/MS method was established to identify multiple compounds in Longjiatongluo capsule,and a reliable HPLC-DAD/ELSD method was developed for the simultaneous anlysis of 6 active ingredients,including dioscin,pseudoprotodioscin,protodioscin,syringin,chlorogenic acid and isofraxidin.First,a qualitative method was established using a Kromasil C18 column(250 mm¡Á4.6 mm£¬5 ¦Ìm) with acetonitrile-water(with 0.1% formic acid) as mobile phase by gradient elution.As a result,18 compounds were identified based on MS information,and 7 of them were newly found in the preparation.Afterward,the same column and mobile phase were employed to determine the 6 active ingredients of the identified compounds.The quantitative method was developed by another gradient elution program.Dual wavelengths(syringin,254 nm;chlorogenic acid,isofraxidin,350 nm) were selected to give maximal specific detection using a diode array detector.What¡¯s more,three steroidal saponins including dioscin,pseudoprotodioscin and protodioscin were detected with evaporative light scattering detector at an evaporation temperature of 100 ¡æ,a gas flow of 1.8 L/min and a nebulization temperature of 40 ¡æ.Good linearities of six analytes were obtained in certain concentration with correlation coefficients of 0.999 6-0.999 9,and the average recoveries(n=9) were in the range of 99.9%-100.6% with RSDs of 0.90%-1.4%.Experimental results showed that the RSDs of precision and repeatability of the six analytes were both less than 2%,and the RSD of stability of the sample in 24 h was also less than 2%.With accuracy,reliability and high repeatability,this method could be used to control the multi-compound quality of Longjiatongluo capsule.