RP-HPLC法测定人血清中维生素C的含量

目的: 建立反相高效液相色谱法测定人血清中维生素C含量。 方法: 采用SHIMADZU VP-ODS色谱柱(4.6 mm×250 mm,5 μm),流动相为30 mmol·L^-1磷酸二氢钾溶液-甲醇(88∶ 12,pH 3.0),流速0.8 mL·min^-1,检测波长243 nm,进样量20 μL,外标法定量。 结果: 维生素C与人血清中其他成分分离良好。维生素C浓度在0.5~15.0 mg·L^-1范围内呈良好的线性关系,r=0.9999。高、中、低3个浓度的日内RSD分别为0.5%,5.0%,2.1%;日间RSD分别为5.0%,6.5%,4.8%;平均回收率分别为93%,88%,84%。 结论: 采用反相高效液相色谱法测定人血清中维生素C的含量,样品处理简便,结果准确可靠。该方法可用于大量血清样品中维生素C的含量测定。

Objective: To establish an RP-HPLC method for the determination of vitamin C in human serum. Methods: The chromatographic column was SHIMADZU VP-ODS(4.6 mm×250 mm,5 μm),the mobile phase was 30 mmol·L^-1KH₂PO₄ solution-methanol(88∶ 12,pH 3.0) at the flow rate of 0.8 mL·min^-1,detection wavelength was 243 nm,and injection volume was 20 μL,using external standard method. Results: Vitamin C was separated well from other ingredients in serum.The linear range was 0.5-15.0 mg·L^-1(r=0.9999) for vitamin C.The intra-day RSDs of high,medium and low concentration were 0.5%,5.0%,2.1%,the inter-day RSDs were 5.0%,6.5%,4.8%,and the average recoveries were 93%,88%,84%,respectively. Conclusion :The RP-HPLC method proposed for the determination of vitamin C in human serum is simple,reliable,accurate,and can be applied for the determination of a great number of samples.