A sensitive HPLC–MS/MS method for the simultaneous determination of anemoside B4, anemoside A3 and 23‐hydroxybetulinic acid: Application to the pharmacokinetics and liver distribution of Pulsatilla chinensis saponins

Abstract

Pulsatilla chinensis saponins, the major active components in the herb, have drawn great attention as potential hepatitis B virus infection and hepatoma treatments. Here, a sensitive and accurate HPLC-MS/MS method was established for simultaneous determination of three saponins - anemoside B4, anemoside A3 and 23-hydroxybetulinic acid - in rat plasma and liver, and fully validated. The method was successfully applied to a pharmacokinetics and liver distribution study of P. chinensis saponins. Consequently, 23-hydroxybetulinic acid, with an extremely low content in the P. chinensis saponins, exhibited the highest exposure in the liver and in sites before and after hepatic disposition, namely, in the portal vein plasma and systemic plasma, followed by anemoside B4, which showed the highest content in the herb, whereas anemoside A3 displayed quite limited exposure. The hepatic first-pass effects were 71% for 23-hydroxybetulinic acid, 27% for anemoside B4 and 37% for anemoside A3, corresponding to their different extents of liver distribution. To our knowledge, this is the first investigation on the liver first-pass effect and distribution of P. chinensis saponins to date. These results also provide valuable information for the understanding of the pharmacological effect of P. chinensis saponins on liver diseases.

Advancements and challenges in pharmacokinetic and pharmacodynamic research on the traditional Chinese medicine saponins: a comprehensive review

“…Saikosaponin B4 had the highest initial concentration in the herbal medicine, followed by Hederagenin, which has relatively limited exposure. The hepatic first-pass effect was 23% for 23-hydroxybetulinic acid, 27% for saikosaponin B4, and 3% for Hederagenin, reflecting their varying degrees of hepatic distribution ( Guo et al, 2018 ).…”

Integrated transcriptomic and metabolomic analyses revealed the molecular mechanism of terpenoid formation for salicylic acid resistance in Pulsatilla chinensis callus

“…The terpenoid and monomers in P. chinensis are widely adapted to clinical and veterinary treatment for their anticancer activity (Kang et al, 2019;Huihui Song et al, 2021). Pulsatilla saponins B3, BD, B4, B7, B9, B10, B11, A3, D, PSA, R13 and other components have been widely reported for their significant pharmacological activities (Xu et al, 2013;Liu, 2014;Liang, 2016;Guo et al, 2018;Yingying Luo et al, 2018;Kang et al, 2019;Yaru Cui et al, 2019;Zhang et al, 2021). Yingying Luo isolated and identified the total saponins of P. chinensis and the isolated monomers to study anti-tumor activity in vitro, and found that monomers of B3, B7, B9 and B11 could significantly inhibit tumor proliferation (Luo, 2014).…”

Anemoside B4 Exerts Hypoglycemic Effect by Regulating the Expression of GLUT4 in HFD/STZ Rats

“…The high content of B4 in the saponins has been used as a quality control marker for pulsatilla. The previous studies of B4 mainly focused on its composition identification, biotransformation, and tissue distribution [ 1 , 2 , 3 , 4 ]. The updated study in recent years reported that B4 has anti-inflammatory, immunomodulatory, antiapoptosis, and autophagy-enhancement effects [ 5 , 6 ].…”