科研之友
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摘要
Purpose: To develop and to validate a simple but sensitive method for determination of vitamins D-3 and K-1 in rat plasma. Methods: The sample treatment included protein precipitation by cold acetonitrile, evaporation, reconstitution with methanol and filtration. The chromatography conditions included Xterra RP18 3.5 mu m 4.6 x 100 mm column at ambient temperature and mobile phase consisting of methanol/water (93/7, v/v) at 0.5 mL/min flow rate. Vitamin D-3 and probucol were detected at 265 nm and vitamin K-1 at 239 nm. Rats were administered intravenously by 0.1 mg/kg of vitamin D-3 or K-1 and the blood samples were withdrawn pre-administration and at pre-determined time points post-administration. The pharmacokinetic analysis was performed using a non-compartmental approach. Results: The calibration curves in rat plasma were linear up to 5000 ng/mL for both vitamins. The limit of quantification (LOQ) was 20 ng/mL for vitamin D-3 and 40 ng/mL for K-1. Inter-and intra-day precision and accuracy were below 15%. The pharmacokinetic parameters of vitamin D-3 following intravenous administration were: AUC(0-infinity) = 11323 +/- 1081 h x ng/mL, V-d = 218 +/- 80 mL/kg, CL = 8.9 +/- 0.8 mL/h/kg, t(1/2) = 16.8 +/- 5 h; and of vitamin K-1: AUC(0-infinity) = 2495 +/- 297 h x ng/mL, V-d = 60 +/- 24 mL/kg, CL = 40.5 +/- 5.1 mL/h/kg, t(1/2) = 1.1 +/- 0.5 h. Conclusion: The developed HPLC-UV assay is a simple and sensitive method for the determination of vitamins D-3 and K-1 in rat plasma. A higher dose of vitamin K-1 should be used in future studies for accurate estimation of pharmacokinetic parameters. The data show the suitability of the assay for pharmacokinetic studies in rats.
- 出版日期2014-3
