不同炮制方法对延胡索中有效成分含量的影响

不同炮制方法对延胡索中有效成分含量的影响

来源：中国论文下载中心 [ 11-05-04 16:59:00 ] 编辑：studa20

作者：曹柳,窦志英,王萍,田永亮

【摘要】目的比较不同炮制方法对延胡索中3种有效成分含量的影响。方法将鲜延胡索、水煮延胡索和市售的延胡索药材分别经醋煮、醋炙和酒炙后,分别采用水超声法和甲醇热回流法进行提取,采用反相高效液相色谱法测定3种有效成分的含量。色谱柱为Agilent

TC-C18(250 mm×4.6 mm,5 μm),流动相：乙腈-0.1%磷酸溶液(三乙胺调pH 5.3) 为28∶72,流速：1 mL/min,柱温：35 ℃,检测波长：280 nm。结果延胡索经炮制后水提液中生物碱含量均有所提高,醇提液中生物碱含量也有上升的趋势；延胡索鲜品直接经醋炙、醋煮、酒炙处理后,去氢紫堇碱含量较高,延胡索在产地水煮后延胡索乙素含量较高。结论本研究建立了同时检测延胡索中3种有效成分的检测方法,该方法简便、可靠、重复性好。根据临床需要,延胡索炮制工艺可采用鲜品水煮后再炮制或产地加工和炮制一体化的两种操作方式。

【关键词】延胡索；原阿片碱；去氢紫堇碱；延胡索乙素；高效液相色谱法；炮制

Abstract：Objective To explore the effect of different processes on the contents of active ingredients in Rhizoma Corydalis. Methods Processes of vinegar-boiling, vinegar-frying and alcohol-frying were used to fresh corydalis, boiled corydalis and purchased corydalis. Then the method of extraction by ultrasound with water and by reflux with methanol were used to extract the sample. The contents of Protopine, Tetrahydropalmatine and Dehydrocorydaline in corydalis was determined by RP-HPLC. Agilent TC-C18 (25 mm×4.6 mm, 5 μm) column was used at

35 ℃, with acetonitrile-0.1% phosphonic acid (pH 5.3 with triethylamine) (28∶72) as mobile phase. The flow rate was 1 mL/min and detection wavelength was set at 280 nm. Results In the sample extracted by water, the contents of active ingredients in processed products were higher than in corydalis. And in the sample extracted by methanol, it also has the tendency that the contents of active ingredients in processed products were higher than in corydalis. The content of Dehydrocorydaline in fresh corydalis which was processed by vinegar-boiling, vinegar-frying and alcohol- frying was high. The content of Tetrahydropalmatine in purchased corydalis which was processed by water-boiling at production place was high. Conclusion The method of determining active ingredients in Rhizoma Corydalis was established. It was convenient and the result was accurate. The process technology that fresh corydalis was boiled by water and then processed or fresh corydalis was processed in production place can be used.

Key words：Corydalis；Protopine；Dehydrocorydaline；Tetrahydropalmatine；HPLC；process

延胡索为罂粟科植物延胡索Corydalis yanhusuo W.T.Wang的干燥块茎,具有活血、利气、止痛功效,临床用于治疗胸胁、脘腹疼痛、经闭痛经、产后瘀阻、跌扑肿痛等症[1],主产于浙江。延胡索中主要有效成分为生物碱,生物碱在药材中多数以结合态的生物碱盐存在,少数碱

性极弱的以游离态形式存在。延胡索的药理作用表现为镇痛、催眠、降低冠脉阻力及增加血流量等,另外亦对心血管系统有较好的药理活性。延胡索多炮制后入药,主要炮制方法为醋煮、醋炙和酒炙。本试验采用水超声提取法和甲醇加热回流提取法分别提取总生物碱盐和总生物碱,通过高效液相色谱法比较了不同炮制方法对延胡索中延胡索乙素、原阿片碱和去氢紫堇碱含量的影响,为优化延胡索炮制工艺和阐明炮制机理奠定基础。

1 仪器与试药

烘箱DHG78-1(湖北省黄石市医疗器械厂)；日本岛津HPLC系统(SCL-10AVP HPLC Pump；SPD-ML0AVP二级管阵列检测器)；自动进样系统；超声波清洗器AS20500AT(天津奥特赛恩斯仪器有限公司)。甲醇、乙腈为色谱纯(天津市协和昊鹏色谱科技有限公司)；其余试剂均为分析纯；水为超纯水。延胡索乙素(批号0726-200409,中国药品生物制品检定所)；去氢紫堇碱(批号043-27611,日本和光纯药工业株式会社)；原阿片碱(批号0853-200201,中国药品生物制品检定所)。

鲜延胡索药材产自浙江东阳；市售延胡索购于天津中新药业集团股份有限公司药材分公司,产自浙江。经本校中药鉴定教研室马琳教授鉴定为罂粟科植物延胡索Corydalis yanhusuo W.T.Wang的块茎。

2 方法与结果

2.1 样品的制备

醋煮延胡索：取延胡索适量,加入20%的醋,加适量水没过药面,浸润4 h,煮干[2]。

醋炙延胡索：取延胡索适量,加入20%的醋,加适量水没过药面,浸润4 h,文火炒干[3]。

酒炙延胡索：取延胡索适量,加入10%的酒,加适量水没过药面,浸润4 h,文火炒干。

鲜延胡索：将新鲜的延胡索经过晾晒后(含水量约为20%),除去泥土,洗净,按照上述各炮制方法进行炮制。

水煮延胡索：将新鲜的延胡索经过晾晒后(含水量约为20%),加水煮至内无白心,取出,干燥。按照上述各炮制方法进行炮制。

市售延胡索：直接按照各炮制方法进行炮制。

2.2 延胡索乙素、去氢紫堇碱和原阿片碱的含量测定

2.2.1 色谱条件与系统性试验

色谱柱为Agilent TC-C18 (250 mm×4.6 mm,5 μm),美国安杰伦公司；流动相：乙腈-0.1%磷酸溶液(三乙胺调pH 5.3)＝28∶72,流速为1 mL/min,柱温为35 ℃,检测波长为280 nm。在此条件下,理论塔板数按延胡索乙素峰、去氢紫堇碱峰和原阿片碱峰计算均不低于3 000。空