高效液相色谱法测定尿样中维生素C、B1和B2

目的:建立高效液相色谱(HPLC)法同时测定尿样中维生素C、B1和B2浓度.方法:以Phenomenex Synergi HydroRP80A(250 mm× 4.6 mm,4μm)为色谱柱,甲醇为流动相A,50 mmol/L乙酸铵溶液(含5mmol/L辛基磺酸钠)为流动相B,梯度洗脱；检测波长:266nm；流速:0.8mL/min;柱温:30℃；尿样经离心过滤后直接进样5μL至HPLC检测.结果:维生素C、B1和B2的线性范围分别为1.3～250.0,0.3～50.0和0.1～25.0 μg/mL,相关系数均大于0.99;回收率分别为98.1％,98.8％和98.7％.结论:该法可用于尿样中3种维生素的同时检测,简便快速,结果准确.

Simultaneous Determination of Vitamin C, B1 and B2 in Urine by High-performance Liquid Chromatography

Objective:To establish an high-performance liquid chromatography(HPLC)method for simultaneous determination of VC,VB1 and VB2 in urine.Method:Phenomenex Synergi Hydro-RP 80A column(250 mm×4.6 mm,4 μm)was used,methanol as mobile phase A and 50 mmol/L ammonium acetate(containing 5 mmol/L 1-octanesulfonic acid sodium salt)as mobile phase B,gradient elution,detector wavelength:266 nm,flow rate: 0.8 mL/min,column temperature:30 ℃.Urine sample was centrifuged directly and the supernatant was filtered,a 5-μL aliquot was injected into the HPLC for analysis.Results:The liner ranges of VC,VB1 and VB2 were 1.3 ～ 250.0,0.3 ～ 50.0 and 0.1 ～ 25.0 μg/mL with correlation coefficients more than 0.99,respectively.The average recoveries were 98.1%,98.8% and 98.7%,respectively.Conclusion:This method is rapid,convenient and accurate for simultaneous determination of three vitamins in urine.