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反相高效液相色谱法测定血浆中的辅酶Q10
Determination of Coenzyme Q10 in Human Plasma by Reversed-Phase High Performance Liquid Chromatography
【摘要】 建立了一种检测血浆中辅酶Q10含量的高效液相色谱法。血浆经甲醇脱脂蛋白后,以正己烷萃取,萃取液依次经硅胶柱净化、C18柱固相萃取,再进行高效液相色谱分析。色谱柱为HypersilODS2柱(5μm,150mm×4 6mmi d ),以异丙醇 甲醇(体积比为45∶55)溶液作流动相,辅酶Q9作内标,检测波长为275nm。在0 1~50 0mg/L质量浓度范围内,辅酶Q10与辅酶Q9的峰面积比与相应CoQ10的质量浓度呈良好的线性关系(r2=0 999),血浆中辅酶Q10的检测限为0 03mg/L(S/N=3),加标回收率为96%~98%,方法重现性好(RSD<3%)。用此法测定正常人血浆中辅酶Q10的含量水平,结果令人满意。
【Abstract】 An improved high performance liquid chromatographic (HPLC) method for the determination of coenzyme Q10(CoQ10) in human plasma has been developed. CoQ10 was dissociated from lipoproteins in plasma by treatment with methanol and extracted with nhexane, then cleanedup on silica gel and C18 solidphase extraction cartridges and finally analyzed by HPLC. The HPLC conditions were: A Hypersil ODS2 column (5 μm, 150 mm×46 mm id) as analytical column, isopropanolmethanol (45∶55, v/v) at a flowrate of 1 mL/min as mobile phase, coenzyme Q9 (CoQ9) as internal standard and UV 275 nm as detection wavelength. The linearity between concentration and peak area ratio (CoQ10/CoQ9) was in the range from 01 to 500 mg/L (r2=0999) and the minimum detectable CoQ10 plasma level was 003 mg/L (S/N=3). The recoveries ranged from 96% to 98%. The method has a good reproducibility with RSD less than 3%. Finally, the method was applied to determine CoQ10 concentrations in healthy human plasma.
【Key words】 high performance liquid chromatography; solid-phase extraction; coenzyme Q10 (CoQ10); plasma;
- 【文献出处】 色谱 ,Chinese Journal of Chromatography , 编辑部邮箱 ,2003年06期
- 【分类号】O657.72
- 【被引频次】10
- 【下载频次】265