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| Rapid Quantitative Analysis of Aconitine-type Alkaloids and Ginsenosides in Shenfu Injection by Column Switching Liquid Chromatography |
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| DOI： |
| KeyWord:column switching liquid chromatography online SPE benzoylmesaconine aconitine gensinoside Ro Shenfu injection |
| Author | Institution |
| ZHANG Yan-hai，LI Ling，ZHANG Da-wei，LU Rui |
1. Agilent Technologies （China） Co. LTD，Shanghai ，China; 2. Heilongjiang Agricultural Reclamation General Hospital，Harbin ，China |
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| Abstract: |
| A column switching liquid chromatography was developed for the analysis of aconitine-type alkaloids including aconitine（AC），mesaconitine（MA），hypaconitine（HA），benzoylaconine（BAC），benzoylmesaconine（BMA），benzoylhypaconine（BHA） and nine ginsenosides in Shenfu injection（SFI）．Aconite-type alkaloids were enriched on an online SPE column by the interaction of strong cation exchange，and purified by the optimized eluting program．Eluting conditions were confirmed through investigating the transferring solvent and eluting time．A Zorbax SB－C18 was chosen as the analytical column，with acetonitrile and ammonium acetate as mobile phases．Meanwhile，an EC－C18 belonged to core-shell particles was used to analyze ginsenoside in SFI．Through the investigation of column oven and gradient program，the optimized conditions were confirmed．The matrix interference was removed to improve the relative response in chromatogram by column switching．The analytical processes for alkaloids and ginsenosides were switched automatically via valve switching．The satisfactory resolutions could be achieved for the aconitine-type alkaloids and ginsenosides in SFI sample．The methodology investigation showed good linearity（r2 > 0.999），precision（RSD < 2.0%） and better reproducibility（RSD < 2.0%）．The limits of detection（LODs） for six target alkaloids were from 4.0 to 8.2 ng/mL．The average recoveries for six target alkaloids ranged from 95.1% to 98.6%．For nine ginsenosides，the recoveries were from 91.7% to 104%．It was an effective way to remove a large amount of matrix interference by the constructed online SPE method based on column switching LC，which could complete the rapid quantitation of three monoester alkaloids and nine active ginsenosides in SFI．The proposed method also realized the limit inspection of three diester alkaloids，and it could be used in practical quality evaluation on SFI． |
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