Assessment of baicalin in mouse blood by monoclonal antibody‐based icELISA |
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| Authors: | Yan Zhao Hui Kong Ye Sun Huibin Feng Yue Zhang Xin Su Huihua Qu Qingguo Wang |
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| Affiliation: | 1. School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing, China;2. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, China;3. Center of Scientific Experiment, Beijing University of Chinese Medicine, Beijing, China |
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| Abstract: | An indirect competitive enzyme‐linked immunosorbent assay (icELISA) based on monoclonal antibaodies (MAb) was recently developed. This new method displays high sensitivity and accuracy, and is especially suitable for pharmacokinetic studies in small laboratory animals. This study aimed to develop an icELISA procedure for baicalin (BAL) quantitation in blood. We successfully developed the icELISA and applied in pharmacokinetic assays of Gegen Qinlian Decoction in mice. A linear correlation was obtained for BAL concentrations in the range from 34.69 to 2220.00 µg/L. The regression equation was y = 1.5557 ? 0.4028log(C) with a correlation coefficient of 0.9936. Precision and accuracy of the icELISA method were evaluated by the variations between replicates from well to well (intra‐assay) and plate to plate (inter‐assay). The values obtained for these parameters were within the normal range (<15%). The recovery rates ranged from 98.93 to 126.78%, meeting the requirements for biological samples. Stability studies showed that BAL sample solutions were intact for 1 h, enough time for UV detection. However, long‐term storage and especially freeze–thaw procedures were detrimental to BAL. The pharmacokinetic parameters derived from mouse experiments were as follows: area under the curves from time 0 to 48 h, 1876.15 ± 1108.14 mg h/L; mean maximum blood concentrations, 101.09 ± 31.53 mg/L; time of maximum concentration, 3.58 ± 2.88 h; mean residence time, 79.30 ± 61.21 h. Copyright © 2014 John Wiley & Sons, Ltd. |
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| Keywords: | icELISA baicalin pharmacokinetics mouse traditional Chinese medicine |
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