中药罗仙子抗动脉粥样硬化活性部位氨基酸HPLC指纹图谱分析

Study on HPLC fingerprint of amino acids in anti-atherosclerotic active extract of Musca domestica

目的：构建中药罗仙子抗动脉粥样硬化活性部位氨基酸指纹图谱,为明确其氨基酸组成及建立其质量控制方法提供实验依据。方法用2,4-硝基氟苯柱前衍生法建立罗仙子抗动脉粥样硬化活性部位水解氨基酸的HPLC指纹图谱,优化衍生条件。结果衍生剂量为1%（体积分数）2,4-二硝基氟苯（2,4-dinitrofluorobenzene,DNFB ）乙腈溶液1.0 mL、衍生温度为60℃、衍生时间为60 min时,衍生化完全；样品水解氨基酸中谷氨酸和精氨酸的质量分数较高,均大于5%；HPLC指纹谱图共确定24个共有峰,10批样品相似度在0．99-1．00之间。结论本方法稳定、可靠,重复性好,明确了罗仙子抗动脉粥样硬化活性部位的氨基酸组成,并初步建立了该部位的质量控制方法。

Objective To study on the HPLC fingerprint of amino acids in anti-atherosclerotic active extract from Musca domestica and provide experimental evidence for the amino acid composition and quality control. Methods 2,4-dinitrofluorobenzene（ DNFB） precolumn derivatization was used and the conditions were optimized. Results When the sample was derivatized by 1.0 mL 1%（φ） DNFB acetonitrile solution under 60℃ for 60 min,it could be completely derivatized. The glutamic acid and arginine were higher than other amino acids and their contents were both more than 5%. 24 common peaks in the HPLC fingerprint of anti-atherosclerotic active extract of Musca domestica were identified and the similarity for 10 bathes of samples was among 0.99-1.00. Conclusion Good fingerprints were obtained and the amino acids of anti-atherosclerotic active extract of Musca domestica were defined and the quality control methods were initially established.