Abstract
A sensitive and reliable HPLC-MS/MS method has been developed and validated for simultaneous determination of eleven bioactive compounds (rhein, emodin, stilbene glycoside, liquiritin, ononin, verbascoside, gallic acid, schisandrin, liquiritigenin, glycyrrhizic acid, and isoliquiritigenin) in rat plasma after oral administration of Tongmai Yangxin Pill. The collected plasma samples were prepared by liquid-liquid extraction with ethyl acetate after acidification. Eleven compounds were separated on a CORTECS™ C18 column with mobile phases consisting of 0.1% formic acid in deionized water and acetonitrile. The flow rate was 0.3 mL/min. The detection was performed on a tandem mass system with an electrospray ionization (ESI) source in both positive and negative ionization using multiple-reaction monitoring (MRM) mode. The calibration curves were linear over the range of 8-2000 ng/mL for glycyrrhizic acid; 4-1000 ng/mL for liquiritin; 0.8-200 ng/mL for emodin, gallic acid, ononin, schisandrin, and stilbene glycoside; 0.4-100 ng/mL for isoliquiritigenin, liquiritigenin, rhein, and verbascoside, respectively. The intra- and interday precision of the analytes were less than 9.3% and 8.5%. The intra- and interday accuracy were in the range of -14.0% to 10.3% and -6.5% to 9.6%. Meanwhile, the extraction recovery of the analytes in plasma samples ranged from 85.2% to 109.1% and matrix effect from 89.2% to 113.4%. The developed method was successfully applied to the pharmacokinetics of eleven bioactive compounds in rat plasma after oral administration of Tongmai Yangxin Pill prescription.
Highlights
Traditional Chinese Medicine (TCM) is a precious treasure of nature
Emodin, stilbene glycoside, liquiritin, ononin, verbascoside, gallic acid, schisandrin, liquiritigenin, glycyrrhizic acid, isoliquiritigenin, and icariin were purchased from Chengdu Must Bio-Technology Co., Ltd. (Chengdu, China)
The final concentrations of the series analytes were at the range of 8-2000 ng/mL for glycyrrhizic acid; 4-1000 ng/mL for liquiritin; 0.8-200 ng/mL for emodin, gallic acid, ononin, schisandrin, and stilbene glycoside; and 0.4-100 ng/mL for isoliquiritigenin, liquiritigenin, rhein, and verbascoside
Summary
Traditional Chinese Medicine (TCM) is a precious treasure of nature. TCMs have been used in clinical for thousands of years and attracting rising attentions due to the treatment of divers diseases successfully with minimum side effects [1,2,3]. The prescription consists of eleven herbs including Radix rehmanniae, Caulis spatholobi, Radix glycyrrhizae, Ramulus cinnamomi, Radix ophiopogonis, Radix polygoni multiflori preparata, Asini corii colla, Fructus schisandrae, Radix codonopsis, Capapax et Plastrum testudinis, and Fructus jujubae [6]. Modern pharmacological studies show that TMYX has a significant effect on heart disease. The 40 components including 2 from Radix rehmanniae, 10 from Radix codonopsis, 2 from Radix ophiopogonis, 2 from Ramulus cinnamomi, 19 from Radix glycyrrhizae, 2 from Radix polygoni multiflori preparata, 5 from Caulis spatholobi, 1 from Fructus jujubae, and 1 from Fructus schisandrae, some of which are overlapped. A reliable and sensitive HPLC-MS/MS method was first developed and applied to the pharmacokinetic study of 11 bioactive components including rhein, emodin, stilbene glycoside, liquiritin, ononin, verbascoside, gallic acid, schisandrin, liquiritigenin, glycyrrhizic acid and isoliquiritigenin in rats after oral administration of TMYX. The pharmacokinetic characteristics of the main chemical components of TMYX in rats were revealed, which would provide a theoretical basis for use of TMYX in clinical
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