反相高效液相色谱-柱前衍生化法测定复方板蓝根颗粒中氨基酸的含量

目的:建立复方板蓝根颗粒中指标性成分精氨酸和脯氨酸的RP-HPLC柱前衍生化法.方法:采用2,4-二硝基氟苯柱前衍生化,将氨基酸在碱性条件下衍生后直接进样测定.Sinochrom ODS-BP(250 mm×4.6 mm,5μm)色谱柱,醋酸钠缓冲溶液(pH6.4)-乙腈(850:150)为流动相,检测波长为360 nm,外标法计算含量.结果:精氨酸和脯氨酸的线性范围分别为0.627～5.016μg和0.874～7.000μg.精氨酸和脯氨酸的平均回收率分别为97.7%(RSD=2.0%)和97.7%(RSD=1.5%).结论:本法简便、准确、可靠,适用于复方板蓝根颗粒的质量控制.

Determination of Amino Acids in Fu Fang Banlangen Granulae by a Pre-column Derivatization RP-HPLC Method

OBJECTIVE:To establish a pre-column derivatization reversed-phase high-performance liquid chromatographic method for the determination of amino acids, which was known as the characteristic constituents of Fu Fang Banlangen Granulae. METHODS: We determined the amino acids in alkalescence by a pre-column derivatization of DNFB. NaAc buffer (pH 6.4)-Acetonitrile (850:150) was used as the mobile phase on a SinoChrom ODS-BP column. The detector was operated at 360 nm. RESULTS: The method was carried out over the range of 0.627 - 5. 016 microg for Arg and 0.874 - 7.000 microg for Pro. The recoveries were 97.7% with the RSD = 2.0% for Arg and 97.7% with the RSD = 1.5% for Pro. CONCLUSIONS: The method has good accuracy and repeatability and it can be used for the quality control of Fu Fang Banlangen Granulae.