摘要
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 develop and validate a simple HPLC-UV method for the determination of coenzyme Q10 and to investigate the concentration-time curve of coenzyme Ql0 in human plasma before and after multiple doses administration. METHODS Coenzyme Q10 inblank plasma was decomposed by light and the plasma was applied as the blank background correction. Coenzyme Q10 in plasma was extracted with n-hexane-ethanol （9：1 ） , and then the separation was performed on a Diamonsil C is column （4. 6 mm × 200 mm, 5 um） with a mixture of methanol-ethanol （ 10：90 v/v） as mobile phase. The internal standard was vitamin K1. The detection wavelength was 275 nm. The concentration-time curve of endogenous coenzyme Q10 was determined in 18 volunteers with the controlled diet for 2 d. The incremental coenzyme Q10 was determined after multiple doses of coenzyme Q10capsules. RESULTS Coenzyme Ql0 was separated from the endogenous substances, the calibration curve was linear in the range of 0. 156 -10. 0 ug . mL-1 with LLOQ = 0. 156 ug . mL-1 （r =0. 999 9）. The human plasma concentration of endogenous eoenzyme Qlo was stable after diet-control for 2 d. The plasma concentration of coenzyme Q10 was increased significantly after multiple doses administration. CONCLUSION The established method was accuracy and sensible for the determination of coenzyme Q10 in human plasma and the settlement of blank background for methodology is reasonable. Muhiple doses administration can be applied to evaluate the bioavailability of coenzyme Q10.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2011年第3期224-228,共5页
Chinese Pharmaceutical Journal
