HPLC异硫氰酸苯酯柱前衍生化测定黄柏及其不同炮制品中17种氨基酸的含量  

Content Determination of 17 Kinds of Amino Acids in Different Processed Products of Phellodendron by HPLC with Pre-column PITC Derivation

作　　者：张凡[1,2,3] 周远征[1,2,3] 赵远[1,2,3] 刘蓬蓬[1,2,3] 徐珊[1,2,3] 贾天柱[1,2,3] 

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]国家中医药管理局中药炮制原理解析重点实验室,辽宁大连116600 [3]辽宁省中药炮制工程技术研究中心,辽宁大连116600

出　　处：《中华中医药学刊》2017年第9期2259-2262,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81274083);国家公益性科研专项项目(201507002)

摘　　要：目的:研究黄柏炮制前后17种氨基酸的含量变化情况进而评价黄柏炮制前后的寒热药性的变化。方法:采用反相高效液相色潽法,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用C18(250mm×4.6 mm,5μm)色谱柱,柱温为43℃;流动相A为0.1 mol·L-1醋酸钠缓冲液(以醋酸调p H 6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1.0 m L·min-1;检测波长为254 nm。结果:17种氨基酸浓度在线性范围内呈良好的线性关系(r=0.996 8~0.999 5);回收率在96.7%~104.26%之间,RSD均小于3.0%。结论:本方法灵敏、准确,具有良好的重复性和稳定性,可用于黄柏中氨基酸的检测。Objective： To study the changes of content of 17 kinds of amino acids in Phellodendron with different processing products. Methods： Use RP-HPLC method with phenyl isothiocyanate（ PITC） derived after derivatization reagent for HPLC analysis. Ecosil C18（ 250 mm × 4. 6 mm,5 μm） column was used. The column temperature was 43 ℃.The mobile phase A was 0. 1 mol·L-1sodium acetate buffer（ adjusted with acetic acid p H 6. 5）-acetonitrile（ 93 ∶ 7）and the mobile phase B was acetonitrile-water（ 4 ∶ 1） by gradient elution. The flow rate was 1. 0 m L·min-1and the detection wavelength was 254 nm. Results： 17 kinds of amino acid concentration in the linear range showed a good linear relationship（ r = 0. 996 8 - 0. 999 5）. The average recovery was between 96. 7% - 104. 26% and RSDs were less than3. 0%. Conclusion： The method is sensitive and accurate,with good reproducibility and stability,and can be used to detect the amino acids in Phellodendron.

关 键 词：黄柏 炮制 氨基酸 中药药性 

分 类 号：R284.1[医药卫生—中药学]