1. 天津大学化工学院制药工程系
2. 天津市生物与制药工程重点实验室
3. 天津世纪天龙药业有限公司
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孙秀婷, 王顺仙, 蒋建兰. 龙加通络胶囊多成分的定性及定量研究[J]. 分析测试学报, 2015,34(1):9-19.
Research on Identification and Determination of Multiple Components in Longjiatongluo Capsule[J]. 2015,34(1):9-19.
采用高效液相色谱-串联质谱技术(HPLC-MS/MS)对龙加通络胶囊多种成分进行定性分析,并建立了其中6种主要药效成分的高效液相色谱-二极管阵列检测/蒸发光散射检测联用(HPLC-DAD/ELSD)定量分析方法。定性分析方法采用Kromasil C18(250 mm×4.6 mm,5 μm)色谱柱进行分离,以乙腈-0.1%甲酸溶液为流动相进行梯度洗脱。定性分析结果根据质谱信息对其中18种化合物进行了成分归属,其中7种为文献未报道的化合物。定量分析方法采用相同的色谱柱和流动相在不同的梯度程序下进行梯度洗脱。DAD采用双波长同时检测:紫丁香苷(254 nm)、异嗪皮啶(350 nm)和绿原酸(350 nm);ELSD检测薯蓣皂苷、原薯蓣皂苷和伪原薯蓣皂苷3种甾体皂苷成分,检测条件为漂移管温度:100 ℃,气体流速:1.8 L/min,雾化器温度:40 ℃。定量分析结果显示,6种成分在各自的检测范围内线性关系良好,相关系数(r)为0.999 6~0.999 9;6种成分的平均加标回收率为99.9%~100.6%,相对标准偏差(RSD)为0.90%~1.4%。精密度、重复性以及24 h稳定性考察中,6种成分的RSD均小于2%。该方法准确可靠、重复性好,可用于龙加通络胶囊多成分的质量控制。
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×4.6 mm,5 μm) 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 ℃,a gas flow of 1.8 L/min and a nebulization temperature of 40 ℃.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.
高效液相色谱-串联质谱多成分含量测定高效液相色谱-二极管阵列检测/蒸发光散射检测龙加通络胶囊
HPLC-MS/MSmulti-component determinationHPLC-DAD/ELSDLongjiatongluo capsule
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