Establishment and application for simultaneous determination method of atorvastatin and its active/toxic metabolites in rat plasma
- VernacularTitle:同时测定大鼠血浆中阿托伐他汀及其活性和毒性代谢产物的方法建立与应用
- Author:
Yuchen SONG
1
;
Lin YANG
2
;
Mingqian SUN
1
;
Changying REN
1
;
Jianxun LIU
1
;
Ying ZHANG
1
Author Information
1. Beijing Key Laboratory of Pharmacology of Chinese Materia Medica，Institute of Basic Medical Sciences of Xiyuan Hospital，China Academy of Chinese Medical Sciences，Beijing 100091，China
2. National Clinical Research Center for Chinese Medicine Cardiology，Xiyuan Hospital，China Academy of Chinese Medical Sciences，Beijing 100091，China
- Publication Type:Journal Article
- Keywords:
atorvastatin calcium;
LC-MS/MS;
hydroxy atorvastatin;
atorvastatin lactone;
pharmacokinetics
- From:
China Pharmacy
2023;34(8):917-922
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To establish a method for simultaneous determination of atorvastatin （ATV） and its active metabolites 2-hydroxy atorvastatin acid （2-HAT）， 4-hydroxy atorvastatin acid （4-HAT） and toxic metabolite atorvastatin lactone （ALT） in rat plasma and apply it for pharmacokinetic study. METHODS LC-MS/MS method was adopted for analysis. The one-step precipitation method was used for processing plasma samples （plasma samples were pretreated by acidification to adjust pH value so as to prevent inversion of configuration）， gradient elution was used to analyze the samples， and the analysis time was 5 min. Electrospray positive ionization was adopted， and positive ion scanning was performed in multi-reaction monitoring. The m/z of quantified ion pairs of ATV and its metabolites such as 2-HAT， 4-HAT and ATL， and internal standard pitavastatin were 559.3→ 440.2， 575.2→440.3， 575.0→440.2， 540.9→448.2 and 422.2→290.0， respectively. After conducting a comprehensive methodological investigation of the analytical method， the concentrations of ATV and its metabolites 2-HAT， 4-HAT，and ATL were determined， and the pharmacokinetic parameters of ATV and its metabolites were calculated using the non- compartment model of WinNonlin 6.1. RESULTS The results of methodological validation showed that endogenous substances in blank plasma did not interfere with the determination of the components to be tested， and the standard curve had a good linear relationship； the lower limits of quantification for ATV， 2-HAT， 4-HAT and ATL were 0.5， 0.5， 0.25 and 0.063 nmol/L， respectively. The precision， accuracy， recovery， matrix effect and stability investigation were all in line with the requirements of biological analysis. Pharmacokinetic analysis showed that after intragastric administration in rats， ATV calcium metabolized rapidly， and was mainly exposed to blood circulation in the form of ATV and 2-HAT， with the lowest concentration of lactone-type metabolites. CONCLUSIONS The established method is precise， rapid and accurate for plasma concentration analysis of ATV and its active/toxic metabolites. The application of the method could help to fully elucidate the pharmacokinetic characteristics of atorvastatin calcium in rats.
