Quantitative determination of saponin components in Huoxue Zhitong Capsules using UHPLC-MS/MS coupled with multivariate statistical analysis
NI Shi-sheng
LIN Yi-fan
HUANG Ming-qing
Abstract：Objective:To develop a method for simultaneous determination of 17 saponin components in Huoxue Zhitong Capsules by ultra-high performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS),coupled with multivariate statistical analysis for quality evaluation of different batches.Methods:A Thermo Fisher Scientific Accucore Phenyl Hexyl column(100 mm×2.1 mm,2.6 μm)was used,with mobile phase consisting of 0.1%formic acid aqueous solution-acetonitrile for gradient elution,at a flow rate of 0.4 mL·min-1,and the column temperature was set at 35℃.Mass spectrometry employed a heated electrospray ionization source(HESI),operating in negative ion mode with multiple reaction monitoring(MRM)scanning.Agglomerative hierarchical clustering analysis,principal component analysis(PCA),and boxplot analysis were used to comprehensively evaluate the differences between Huoxue Zhitong Capsules from different manufacturers.Results:The 17 saponin components showed good linear relationships within the linear range(r≥0.9968),with precision,repeatability,and stability RSD all less than 5.00%.The average recoveries ranged from 93.2%to 108.0%,with RSD of 1.01%to 5.67%.The contents of the 17 saponins in 10 batches of Huoxue Zhitong Capsules,including notoginsenoside R1,Fa,ginsenoside Re,Rg1,Rf,F3,Rg2,Rb1,Rc,F1,Rb2,Rb3,Rd,F2,Rg5,20(S)-ginsenoside Rg3 and 20(R)-ginsenoside Rg3 were 346.2976～730.3066,59.1678～125.9001,189.4788～391.8343,1696.3028～2078.3066,2.4021～3.1435,40.31299～71.5728,37.1285～57.8093,968.6498～1 784.8576,4.5467～6.2157,4.3753～10.1749,10.1996～22.9377,7.5306～12.3707,337.7592～686.3070,4.3891～9.4875,0.0000～11.5466,6.321 1～9.623 1,and 0.000 0～1.426 2 μg·g-1,respectively.Multivariate statistical analysis revealed differences in saponin components among Huoxue Zhitong Capsules produced by different manu-facturers.Even for products from the same manufacturer but different batches,there were certain degrees of variation in the contents of 17 kinds of saponins.Conclusion:The developed analytical method demonstrates good separation efficiency and sensitivity,enabling efficiently quantification of 17 saponin components in Huoxue Zhitong Capsules,providing a scientific basis for quality control optimization and standard improvement.
Keywords：Huoxue Zhitong CapsulesUHPLC-MS/MSsaponinsmultivariate statisticalcontent deter-mination
Publication Date：2026-02-28
Online Publishing Date：2026-08-14(First online date of this platform, not the publication date of the document)
Pages：12( 375-386 )