| RP—HPLC法测定白细胞中LTB4和5-HETE的含量 |
| |
| 引用本文： | 林景瑞,陈晓辉,毕开顺,王灵芝,乔延江.RP—HPLC法测定白细胞中LTB4和5-HETE的含量[J].世界科学技术,2009,11(6):872-875. |
| |
| 作者姓名： | 林景瑞 陈晓辉 毕开顺 王灵芝 乔延江 |
| |
| 作者单位： | 林景瑞,陈晓辉,毕开顺(沈阳药科大学药学院,沈阳,110016);王灵芝,乔延江(北京中医药大学中药信息工程中心,北京,100102) |
| |
| 基金项目： | 北京市科委重点实验室(JD100260652):清热解毒中药有效成分发现研究，国家中医药管理局中医药行业科研专项(200707010):针对病毒性疾病的中药活性发现关键技术研究 |
| |
| 摘 要： | 目的：建立了5-脂氧酶抑制剂筛选模型的RP—HPLC测定方法。方法：采用BravaBDSC18柱（250mm×4．6mm，5μm），流动相为甲醇-0．02％乙酸水溶液，流速为0．8mL·min^-1，5-LOX的主要产物ⅡB4和5-HETE的检测波长分别为275nm和235nm。结果：LTB4和5-HETE分别在2．0～50．0μg·mL^-1（r=0．9999）和5．0～125．0μg·mL^-1（r=1．0000）范围内呈良好的线性关系；回收率分别为100．0％和97．7％（RSD分别为1．1％和1．5％）；日内、日间精密度的RSD分别为0．7％，1．1％和0．3％，1．3％。结论：该法分离效果和重复性均良好，且结果准确可靠，可用于LTB4和5-HETE的检测，为筛选新的具有抗炎活性的中药成分奠定了基础。
|
| 关 键 词： | 高效液相色谱法LTB4 5-HETE 5-脂氧酶抑制剂 |
Determination of LTB4 and 5-HETE in Leukocytes by RP-HPLC |
| |
| Institution: | Lin Jingrui, Chen Xiaohui, Bi Kaishun,Wang Lingzhi, Qiao Yanjiang(School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016 , China ;Research Center of TCM-information Engineering, Beijing University of Chinese Medicine, Beijing 100102, China) |
| |
| Abstract: | This study aimed to develop an efficient RP-HPLC method for determination of 5-1ipoxygenase （5-LOX） inhibitory activity. The content of LTB4 and 5-HETE, the main products of the 5-LOX metabolism pathway of arachidonic acid, was determined by the RP-HPLC method. The RP-HPLC was performed on a Brava BDS C18 column （250mm×4.6mm, 5μm） using a mobile phase consisting of 0.02% acetic acid solution-methanol at 30 ℃ with the flow rate of 0.8mL·min^-1 and detection wavelength at 235nm and 275nm. As a result, the calibration curve was hnear in the range of 2.0-50.0 μg·mL^-1 （r = 0.9999） for determination of LTB4, and 5.0-125.0μg·mL^-1 （r=1.0000） for determination of 5-HETE. The average recoveries of LTB4 and 5-HETE were 100.0% （RSD=1.1%） and 97.7% （RSD=1.5%）, respectively. The intra-day and inter-day RSDs were 0.7%, 1.1% and 0.3%, 1.3%. Being sensitive, accurate and reliable with good reproducibility, this method is available for the screening of new anti-inflammatory active components of Chinese medicine. |
| |
| Keywords: | LTB4 5-HETE |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
