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KMID : 0369820090390040309
Jorunal of Korean Pharmaceutical Sciences
2009 Volume.39 No. 4 p.309 ~ p.314
Validation of an Enzyme-Immunoassay for Calcitriol in Human Plasma and Evaluation of Its Pharmacokinetics after Single-dose in Korean Volunteers
Kim Ye-Tae

Jin Su-Eon
Kim Hyun-Ki
Shin Baek-Ki
Jeong Ui-Hyeon
Kim Chong-Kook
Park Jeong-Sook
Abstract
An enzyme immunoassay (EIA) was validated for quantitation of cacitriol in human plasma. Calcitriol was immunoextracted with immunocapsules, which contain monoclonal antibodies to calcitriol linked to solid phase particles in suspension with a vitamin D binding protein inhibitor. Calcitrol was eluated and the eluates were evaporated under a gentle stream of nitrogen gas. The absorbance of analytes was determined using a microplate reader (reference wavelength 650 nm; measurement wavelength 450 nm). The method was specific and sensitive enough to detect as low as 6.5 pmol/L of calcitriol. Linear calibration range was 6.5-491 pmol/L with correlation coefficient greater than 0.99. The overall accuracy was in the range of 83.8 to 111.2% and precision C.V. (%) 0.99 to 8.47%. The recovery was approximately 100% and stability was confirmed during storage and sample preparation. The pharmacokinetic parameters were calculated by baseline subtraction because calcitriol is an endogenous material. Following oral dose of calcitriol, the mean AUC was 1038539 pmol/Lhr and C of 12863.1 pmol/L was reached at 3.501.07 hr. The mean t of calcitriol was 5.132.10 hr. The present EIA method was successfully applied to study bioavailability after oral administration of 2 g of calcitriol in healthy Korean subjects.
KEYWORD
Calcitriol, Enzyme immunoassay (EIA), Validation, Pharmacokinetics
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