手性色谱法测定L-5-甲基四氢叶酸钙含量

Determination of Calcium L-5-Methyltetrahydrofolate by Chiral Chromatography

建立手性色谱法测定L-5-甲基四氢叶酸钙(L-5-MTHF-Ca)的分析方法,通过方法学验证评价方法的可行性。使用高效液相色谱系统,色谱柱为CHIRALPAK HSA手性色谱柱(4 mm×150 mm,5μm),以磷酸盐缓冲溶液∶乙腈(体积比90∶10)为流动相,流速0.6 mL/min,进样量5μL,柱温30℃,检测波长280 nm。方法学验证结果表明,此方法可很好地分离L-5-MTHF-Ca,分离度达到3.24,并且在0.016~0.100 mg/mL质量浓度范围线性关系良好,相关系数为0.9997,检出限为3.11μg/mL,定量限为4.98μg/mL,精密度0.38%,加标平均回收率103.71%,系统适用性良好,然而,L-5-MTHF-Ca会随时间缓慢降解,需尽快进样分析。方法操作简便,结果准确,可用于各种食品、保健品中L-5-MTHF-Ca含量的测定。

A chiral chromatographic method for the determination of calcium L-5-methyltetrahydrofolate(L-5-MTHF-Ca)was established,and the feasibility of the method was evaluated by methodology validation.Methods high performance liquid chromatography is used.The chromatographic column is CHIRALPAK HSA chiral column(4 mm×150 mm,5μm).The mobile phase is phosphate buffer solution∶acetonitrile(90∶10,V∶V).The flow rate is 0.6 mL/min.The injection volume is 5μL.The column temperature is 30℃and the detection wavelength is 280 nm.The results shows that the method could separate L-5-MTHF-Ca well,the resolution is 3.24,and the linear relationship is good in the concentration range of 0.016~0.100 mg/mL,the correlation coefficient is 0.9997,the detection limit and quantitation limit are 3.11μg/mL and 4.98μg/mL respectively,the precision is 0.38%,and the average recovery is 103.71%.System suitability test results are good,but L-5-MTHF-Ca will slowly degrade over time,so sample analysis as soon as possible.The method is simple,accurate and can be used for the determination of L-5-MTHF-Ca in various foods and health products.