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目的建立人血浆中辅酶Q10的HPLC-UV测定法,考察正常人内源性辅酶Q10的经时变化过程,并研究辅酶Q10胶囊连续给药后血浆中辅酶Q10变化规律。方法无辅酶Q10血浆经低温光照后获得,作为方法学建立的空白背景。血浆样品经正己烷-乙醇=9∶1提取后,由迪马Diamonsil C18柱(4.6 mm×200 mm,5μm)分离,流动相为甲醇-乙醇(10∶90),维生素K1为内标,检测波长为275 nm。18名志愿者在控制饮食2 d后,不同时间点采血并测定血浆中内源性辅酶Q10的变化规律,随后连续给予辅酶Q10胶囊同时间点采血,考察血浆中辅酶Q10浓度的升高情况。结果血浆中杂质不干扰样品峰,线性范围为0.156～10.0μg.mL-1(r=0.999 9),定量下限为0.156μg.mL-1。受试者经控制饮食2 d后血浆中内源性辅酶Q10浓度达到稳定状态,连续给予辅酶Q10胶囊后血浆中辅酶Q10的浓度增加明显。结论新建立的人血浆中辅酶Q10测定法,简单快速,准确度高,空白基质选择合理,处理恰当。连续给药的方法可用于评价辅酶Q10的生物利用度。
OBJECTIVE: To establish a HPLC-UV method for the determination of coenzyme Q10 in human plasma and to investigate the changes over time of the endogenous coenzyme Q10 in normal human and to study the changes of coenzyme Q10 in plasma after coenzyme Q10 capsule administration. Methods Coenzyme Q10 plasma obtained by low temperature light, as a methodological establishment of the blank background. The plasma samples were extracted with n-hexane-ethanol = 9: 1 and separated on a Diamonsil C18 column (4.6 mm × 200 mm, 5 μm). The mobile phase consisted of methanol-ethanol (10:90) The wavelength is 275 nm. Eighteen volunteers took blood at different time points and measured the changes of endogenous coenzyme Q10 in plasma after controlling diet for 2 days. Subsequently, coenzyme Q10 capsule was continuously given to take blood at the same time, and the plasma coenzyme Q10 concentration was investigated. Results The impurities in the plasma did not interfere with the sample peak. The linear range was 0.156 ~ 10.0μg.mL-1 (r = 0.999 9). The lower limit of quantitation was 0.156μg.mL-1. After two days of controlled diet, the concentration of endogenous coenzyme Q10 in plasma reached a steady state, and the concentration of coenzyme Q10 in plasma increased significantly after coenzyme Q10 capsule administration. Conclusion The newly established human plasma coenzyme Q10 assay is simple, rapid and accurate. The selection of blank matrix is reasonable and appropriate. The method of continuous administration can be used to evaluate the bioavailability of coenzyme Q10.