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目的采用化学发光免疫法测定大鼠血清金环蛇抗菌肽(BF-30)的含量,为药代动力学研究提供简单快速的检测手段,初步探讨BF-30体内药代动力学特征。方法建立化学发光免疫分析方法并对方法的基质效应、精密度、准确度、稳定性和稀释效应进行确证,应用该法检测Wistar大鼠血清中BF-30的浓度。结果在大鼠血清中,BF-30在0.5~40 ng/ml浓度范围内Logistic曲线拟合良好,最低定量限为0.5 ng/ml;该法不存在基质效应;日内精密度为8.76%~14.15%、日间精密度为4.91%~11.11%,准确度为-1.27%~-7.71%;样品在-20℃放置13 d、反复冻融2次稳定性良好;储备液在-20℃放置30 d稳定性良好;样品在稀释40倍后无稀释效应。结论该分析方法简便、准确、可靠,能满足大鼠血清中BF-30含量的测定及其临床前药代动力学研究。
OBJECTIVE: To determine the content of antimicrobial peptide (BF-30) in rat serum by chemiluminescence immunoassay and provide a simple and rapid method for the study of pharmacokinetics. The pharmacokinetic characteristics of BF-30 in vivo were preliminarily discussed. Methods The chemiluminescence immunoassay method was established and the matrix effect, precision, accuracy, stability and dilution effect of the method were validated. The concentration of BF-30 in serum of Wistar rats was detected by this method. Results The Logistic curve of BF-30 in the range of 0.5 ~ 40 ng / ml was fitted well to the rat sera with the lowest limit of quantitation of 0.5 ng / ml. There was no matrix effect in this method. The intra-day precision was 8.76% ~ 14.15 %, The precision during the day was 4.91% ~ 11.11% and the accuracy was -1.27% ~ -7.71%. The sample was placed at -20 ° C for 13 days and was repeatedly frozen and thawed twice. The stability was good. The stock solution was stored at -20 ° C for 30 days d Good stability; sample diluted 40 times without dilution effect. Conclusion The method is simple, accurate and reliable and can be used to determine the content of BF-30 in rat serum and its preclinical pharmacokinetics.