摘要
目的 建立测定大鼠血浆中内源性化合物辅酶Q10的浓度的方法,并应用该方法测定辅酶Q10药代动力学血浆样品,考察辅酶Q10的药代动力学特征,比较不同剂型的辅酶Q10的相对生物利用度。方法 血浆样品用乙酸乙酯-异丙醇(1∶9,v∶v)溶液蛋白沉淀处理,以辅酶Q10-d9为内标,用标准加入法和背景扣除法相结合消除空白基质中内源性辅酶Q10的干扰,含0.1%甲酸的甲醇溶液为流动相进行等度洗脱,流速为0.60 mL·min^(-1),色谱柱为ACQUITY UPLC-BEH C18柱(2.1 mm×50.0 mm, 1.7μm),柱温为40℃,进样量为1μL。电喷雾离子源(ESI),正离子模式扫描,多反应监测模式(MRM)检测,离子对分别为m/z 863.75→m/z 197.00(辅酶Q10)、m/z 872.70→m/z 205.90(内标)。测定大鼠血浆中辅酶Q10的浓度,然后用药代动力学软件计算其药代动力学参数。结果 辅酶Q10在80.0~8 000.0 ng·mL^(-1)呈现出良好的线性关系(r^(2)>0.990 2),批内与批间精密度RSD分别在1.15%~8.66%和3.55%~6.07%,批内、批间准确度偏差分别在-2.45%~13.35%和0.06%~8.41%;辅酶Q10的回收率在82.35%~96.26%,内标的回收率在92.01%~98.82%,经内标归一化的基质因子在99.47%~99.65%,基质稳定性的测得结果与理论浓度的偏差在±15%。大鼠灌胃5%辅酶Q10自微乳和10%辅酶Q10微粒后,主要药代动力学参数C_(max)分别为(399.14±105.62),(288.03±80.36)ng·mL^(-1),t1/2分别为(5.80±2.17),(5.90±2.07)h, AUC_(0-t)分别为(8 556.35±2 372.02),(6 728.93±1 447.24)h·ng·mL^(-1),5%辅酶Q10自微乳相对于10%辅酶Q10微粒口服相对生物利用度为127.16%。结论 本研究建立了一种快速、灵敏的方法用于内源性化合物辅酶Q10的大鼠血浆浓度的检测,给其药代动力学研究提供了基础。
Objective To establish a method for the quantification of coenzyme Q10 in rats plasma and to investigate the pharmacokinetic characteristics of Coenzyme Q10 and compare the relative bioavailability of different dosage forms of Coenzyme Q10. Methods The plasma samples were precipitated with ethyl acetate-isopropanol(1∶9, v∶v). Using coenzyme Q10-d9 as internal standard, the interference of endogenous coenzyme Q10 in blank substrate was eliminated by a combination of standard addition method and background subtraction method. The determination was performed on ACQUITY UPLC-BEH C18column(2.1 mm×50.0 mm, 1.7 μm) with mobile phase consisted of 0.1% formic acid-methanol(isocratic elution)at the flow rate of 0.60 mL·min^(-1). The column temperature was 40 ℃,and injection volume was 1 μL. Electrospray ionization(ESI) source, positive ion mode scanning, multi-reaction monitoring mode(MRM) were used to detect the effective components. The ion pairs for quantitative analysis werem/z863. 75→m/z197. 00(coenzyme Q10) andm/z872. 70 →m/z205. 90( internal standard), respectively. The concentration ofcoenzyme Q10 in rat plasma was determined by the above-mentioned method and pharmacokinetic parameters werecalculated by pharmacokinetics software.Results The good linear range of coenzyme Q10 was 80. 0-8 000. 0ng·mL^(-1)(r^(2)> 0. 990 2), the intra-day and inter-day precisions( RSDs) were 1. 15%-8. 66%, 3. 55%-6. 07% and the accuracy bias for intra-and inter-batch were-2. 45%-13. 35%,0. 06%-8. 41%. The extractionrecoveries of coenzyme Q10 ranged from 82. 35% to 96. 26% and the extraction recoveries of internal standard rangedfrom 99. 47% to 99. 65%. The matrix factor normalized by internal standard ranged from 99. 47% to 99. 65%. Thedeviation between the measured results of the stability test and the theoretical values were within ± 15%. Afterintragastric administration of 5% coenzyme Q10 self-microemulsion and 10% coenzyme Q10 microparticles, mainpharmacokinetic parameters were as follows:C_(max)were(399. 14 ± 105. 62) and(288. 03 ± 80. 36) ng·mL^(-1);t1/2were(5. 80 ± 2. 17) and(5. 90 ± 2. 07) h;AUC_(0-t)were(8 556. 35 ± 2 372. 02) and(6 728. 93 ± 1 447. 24)h·ng·mL^(-1). The relative bioavailability of 5% coenzyme Q10 self-microemulsion to 10% coenzyme Q10microparticles was 127. 16%.Conclusion The established method is rapid and sensitive for the determination ofendogenous compound coenzyme Q10 in rats plasma, which provides a way for the study of pharmacokineticsin vivo.
作者
王祥艳
张寅静
朱小勇
樊柏林
陈洪建
向霞
王轶
唐晓荞
WANG Xiang-yan;ZHANG Yin-jing;ZHU Xiao-yong;FAN Bo-lin;CHEN Hong-jian;XIANG Xia;WANG Yi;TANG Xiao-qiao(Hubei Provincial Key Laboratory of Applied Toxicology of Hubei ProvincialCenter.for DiseaseControlandPrevention,Wuhan 430075,Hubei Province,China;Zhejiang NHU Company Ltd.,Shaoxing 312000,Zhejiang Province,China;Oil Crops Research Institute of Chinese Academy of Agriculture Sciences,Wuhan 430075,Hubei Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第3期425-429,共5页
The Chinese Journal of Clinical Pharmacology
基金
湖北省重点研发计划基金资助项目(2020BCA086)
湖北省公共卫生领军人才培养计划基金资助项目[鄂卫通(2021)73号]。
