葛根素注射液—葛根素的测定—高效液相色谱法

葛根素注射液—葛根素的测定—高效液相色谱法 方法名称: 葛根素注射液—葛根素的测定—高效液相色谱法 本方法采用高效液相色谱法测定葛根素注射液中葛根素 应用范围: (CHO)的含量。 21209 本方法适用于葛根素注射液。 供试品制成流动相溶液，进入高效液相色谱仪进行色谱分离，用方法原理: 紫外吸收 检测 工程第三方检测合同工程防雷检测合同植筋拉拔检测方案传感器技术课后答案检测机构通用要求培训 器，于波长250nm处检测葛根素吸收值，计算出其含量。 1. 0.1%枸橼酸溶液 试剂: 2. 甲醇 1. 仪器 1.1 高效液相色谱仪 1.2 色谱柱 十八烷基硅烷键合硅胶为填充剂，理论塔板数按葛根素峰计算不 低于2500，葛根素峰与相邻杂质峰间的分离度应符合要求。 仪器设备: 1.3 紫外吸收检测器 2. 色谱条件 2.1 流动相:0.1%枸橼酸溶液 甲醇=75 25。 2.2 检测波长:250nm 2.3 柱温:室温 1. 量取供试品 精密量取本品适量。 2. 对照品溶液的制备 精密称取适量，加流动相溶液溶解并制成每1mL含50μg的溶 液，摇匀，即得。 试样制备: 3. 供试品溶液的制备 将供试品加流动相溶液溶解并制成每1mL含50μg的溶液，摇 匀，即得。 注:“精密称取”系指称取重量应准确至所取重量的千分之一。“精密量取”系指量取体积 的准确度应符合国家标准中对该体积移液管的精度要求。 分别精密吸取上述对照品溶液与供试品溶液各20μL 注入高效 操作步骤: 液相色谱仪，用紫外吸收检测器，于波长250nm处测定葛根素的吸收 值，计算出其含量。 中华人民共和国药典，国家药典委员会编，化学工业出版社，2005参考文献: 年版，二部，p.689。 problems, in order to immediately dry and dry dry determination, leave no stone unturned, down-to-earth to do a good job security and peace, stability, and strive to realize the mass a sense of security and satisfaction. Staring at the "iron Lattice lattice every Duan lattice. The public security organs Service Development and protect the livelihood of the people, the most direct service is to create a good public security environment, let people have a sense of security, let investors to worry. If a burglary can prevent cases of multiple masses safety feeling low, the masses "iron", means that our work dereliction of duty. Therefore, to enhance the people a sense of security and satisfaction, so that "iron" more and less, is our biggest goal. As long as we earnestly to press incidence, enhance people's sense of security and satisfaction as the starting point and the foothold, will be able to work in the Party committee and government approval to obtain the support of the masses, the vast majority of the civilian police support, will not Continue to make new achievements. Second, we must find a good breakthrough point. In recent years, through to the primary research and development of visit, a sense of security satisfaction survey and public reception of visitors, accept masses to complain etc. channel, the police and the masses reflects the tension of police, theft cases, after the report did not deal with the results, the police slowly, service attitude stiff problems. Analysis the above problems, the breakthrough point is to solve tension of police and individual police service attitude problem. To solve the problems of shortage of police 山东省药品检验所 250012 葛根素注射液为新型血管扩张药，用于冠心病、心肌梗塞、心绞痛的治疗，其主成分葛根素为豆科植物野葛Pueraria lobata(Willd.)Ohwi的干燥根中提取、分离得到的以葛根素为主的异黄酮苷类，其含量测定,1,方法，卫生部药品标准(试行)为紫外分光光度法。在生产和检验过程中发现，按100 %投料，紫外法测定其含量可达标示量的103 %左右，经薄层检查，其原料葛根素含杂质，易导致注射液测定含量偏高。本文采用高效液相色谱法，使葛根素与杂质得到良好分离后，测定葛根素的含量，其方法准确，结果可靠。 1 仪器与试药 TSP高效液相色谱仪(包括P 2000泵，AS 3000自动进样器，UV 2000检测器，IBM 350—450 DX2数据站)。 葛根素对照品(中国药品生物制品检定所提供);对-羟基苯甲醛内标物(山东省药物研究所精制，色谱纯)。 乙醇(优级纯)，重蒸馏水，丙二醇(分析纯)，葛根素注射液(烟台中策药业有限公司提供)。 2 色谱条件 色谱柱:Turner CH，5 μm 4.6 mm×150 mm;流动相:乙醇—水(43?1837 207);流速:1 mL/min;柱温:30 ?;检测波长:250 nm;理论板数按葛根素计，不低于3000，与内标峰分离度大于5。 3 线性关系试验 取对-羟基苯甲醛适量，加流动相制成0.5 mg/mL的内标溶液。取105 ?干燥至恒重的葛根素对照品适量，精密称定，用流动相溶解，制成0.3 mg/mL的溶液，精密取该溶液1、3、5、10、20 mL，分别精密加入内标溶液10 mL，摇匀，取各溶液10 μL进样，记录色谱图(见图1)。以葛根素峰面积和内标峰面积之比Y与浓度X(mg/mL)做线性回归，得回归方程为: -3Y,11.98X+5.673×10 r=0.9991 即葛根素在0.027,0.2 mg/mL浓度范围内线性关系良好。 图1 色谱图 1.葛根素 2.内标 3,6.杂质 problems, in order to immediately dry and dry dry determination, leave no stone unturned, down-to-earth to do a good job security and peace, stability, and strive to realize the mass a sense of security and satisfaction. Staring at the "iron Lattice lattice every Duan lattice. The public security organs Service Development and protect the livelihood of the people, the most direct service is to create a good public security environment, let people have a sense of security, let investors to worry. If a burglary can prevent cases of multiple masses safety feeling low, the masses "iron", means that our work dereliction of duty. Therefore, to enhance the people a sense of security and satisfaction, so that "iron" more and less, is our biggest goal. As long as we earnestly to press incidence, enhance people's sense of security and satisfaction as the starting point and the foothold, will be able to work in the Party committee and government approval to obtain the support of the masses, the vast majority of the civilian police support, will not Continue to make new achievements. Second, we must find a good breakthrough point. In recent years, through to the primary research and development of visit, a sense of security satisfaction survey and public reception of visitors, accept masses to complain etc. channel, the police and the masses reflects the tension of police, theft cases, after the report did not deal with the results, the police slowly, service attitude stiff problems. Analysis the above problems, the breakthrough point is to solve tension of police and individual police service attitude problem. To solve the problems of shortage of police 4 校正因子的测定 取在105 ?干燥至恒重的葛根素对照品约20 mg，精密称定，置100 mL 量瓶中，用流动相溶解并稀释至刻度，精密取该溶液与内标溶液各10 mL 混匀，取10 μL进样，记录色谱图，按对照品与内标的峰面积比计算校 正因子。 5 回收率试验 根据葛根素注射液处方精密称取葛根素对照品并配制成模拟溶液(每2 mL的50 %丙二醇水溶液含葛根素100 mg)，精密取该模拟溶液1 mL，置 250 mL量瓶中，加流动相至刻度，摇匀，精密取该溶液与内标溶液各10 mL，混匀， 取10 μL进样，以内标法计算，平均回收率为99.6 %(n=5)， RSD为0.42 %。 6 样品测定 ,1,精密取本品，按回收率项下方法依法测定、计算，结果与紫外法比较， 见表1。 表1 含量测定结果(标示量%，n=4) 批号 HPLC法 紫外法 9507021 98.0 101.6 9507022 98.2 101.6 9507023 98.3 101.8 7 讨论 ,1,葛根素注射液的含量测定，紫外法测得的含量为在250 nm波长处有紫外吸收的总黄酮的总量(将有紫外吸收的杂质一并计算入含量)，如葛根素提取纯度不好，引起实验误差较大;在本文色谱条件下，HPLC法能很好地将葛根素与其它杂质分离后定量，辅料对测定无任何干扰，正确反映了注射液中葛根素含量。 参考文献 1 卫生部药品标准(试行).WS—257(X—207)—92 problems, in order to immediately dry and dry dry determination, leave no stone unturned, down-to-earth to do a good job security and peace, stability, and strive to realize the mass a sense of security and satisfaction. Staring at the "iron Lattice lattice every Duan lattice. The public security organs Service Development and protect the livelihood of the people, the most direct service is to create a good public security environment, let people have a sense of security, let investors to worry. If a burglary can prevent cases of multiple masses safety feeling low, the masses "iron", means that our work dereliction of duty. Therefore, to enhance the people a sense of security and satisfaction, so that "iron" more and less, is our biggest goal. As long as we earnestly to press incidence, enhance people's sense of security and satisfaction as the starting point and the foothold, will be able to work in the Party committee and government approval to obtain the support of the masses, the vast majority of the civilian police support, will not Continue to make new achievements. Second, we must find a good breakthrough point. In recent years, through to the primary research and development of visit, a sense of security satisfaction survey and public reception of visitors, accept masses to complain etc. channel, the police and the masses reflects the tension of police, theft cases, after the report did not deal with the results, the police slowly, service attitude stiff problems. Analysis the above problems, the breakthrough point is to solve tension of police and individual police service attitude problem. To solve the problems of shortage of police , 葛根素注射液--药物手册 【其他名称】 【药物名称】葛根素注射液 【英文名】Puerarin Injection 【汉语拼音】 【主要成分】 【性状】无色或微黄色的澄明液体。 【药理作用】主要药效学实验，进行了静脉注射对犬急性心肌梗塞范围影响的研究，结果表明:可明显限制实验性心肌梗塞范围，此结果系经心外膜电图、酶、心肌核素及心肌NBT病理染色所证实。一般药理学研究结果表明，临床等效量静脉注射对实验动物(犬)呼吸、心血管和神经系统无明显影响。 小鼠静脉注射LD50为634.3mg/Kg，其95%可信限为583.8,689.2mg/Kg;腹腔注射LD50为1412.2mg/Kg，其95%可信限为1377.8,1448.1mg/Kg。大鼠长期毒性实验结果表明，150、100、50mg/Kg腹腔注射5周无积蓄性毒性，对心、肝、肺、脾、肾、肾上腺及肠等脏器无明显毒性。犬长期毒性实验结果表明，15、30、50mg/Kg/日静脉注射，连续35天，再观察70天，对大小便常规、血常规、SGPT、BUN、血糖均无明显影响，但高剂量组病理检查肝肾组织有不同程度受损，给药部位结缔组织增生、管壁增厚。致突变实验结果显示，无任何致突变活性，说明没有潜在性的致癌和致突变危险性。致畸实验结果表明，对雌性大鼠胚胎及雄性大鼠生殖细胞均无致畸作用。 problems, in order to immediately dry and dry dry determination, leave no stone unturned, down-to-earth to do a good job security and peace, stability, and strive to realize the mass a sense of security and satisfaction. Staring at the "iron Lattice lattice every Duan lattice. The public security organs Service Development and protect the livelihood of the people, the most direct service is to create a good public security environment, let people have a sense of security, let investors to worry. If a burglary can prevent cases of multiple masses safety feeling low, the masses "iron", means that our work dereliction of duty. Therefore, to enhance the people a sense of security and satisfaction, so that "iron" more and less, is our biggest goal. As long as we earnestly to press incidence, enhance people's sense of security and satisfaction as the starting point and the foothold, will be able to work in the Party committee and government approval to obtain the support of the masses, the vast majority of the civilian police support, will not Continue to make new achievements. Second, we must find a good breakthrough point. In recent years, through to the primary research and development of visit, a sense of security satisfaction survey and public reception of visitors, accept masses to complain etc. channel, the police and the masses reflects the tension of police, theft cases, after the report did not deal with the results, the police slowly, service attitude stiff problems. Analysis the above problems, the breakthrough point is to solve tension of police and individual police service attitude problem. To solve the problems of shortage of police 【药代动力学】关于药代动力学的研究，采用放射性氚标记的葛根素观察了静脉给药后的血药浓度变化及其在10种脏器中的药物浓度变化，基于开放型二室模型，计算了葛根素药代动力学参数，并对高、中、低三个剂量组的动力学参数进行了比较，结果表明，小鼠静脉注射葛根素后，随着给药剂量的增加，药物的消除半衰期(t1/2β)依次降低(11.80，10.37，4.65hr)，分布半衰期(t1/2α)依次增加(0.53，0.64，0.67hr);药物在各组织的分布以肝、肾、心脏及血浆中较高;睾丸、肌肉和脾脏次之;并可通过血脑屏障进入脑内，但含量较低。通过对各种参数分析结果说明，在体内以分布广，消除快，不易积蓄为特点。 【适应症】用于冠心病、心肌梗塞、心绞痛。 【用法与用量】稀释后静脉滴注。一日400,600mg，15天为一疗程。 【不良反应】个别病例用药开始时有暂时性腹胀、恶心等消化道反应，继续用药自行消失。 【禁忌症】 【注意事项】为50%丙二醇配制，性微粘稠，取样时注意。同时，遇碱变黄，请勿在稀释时加入碱性药物。 【规格】2ml?0.1g 【贮藏】遮光，密闭，在干燥处保存。 【有效期】暂定二年。 【处方药】是 problems, in order to immediately dry and dry dry determination, leave no stone unturned, down-to-earth to do a good job security and peace, stability, and strive to realize the mass a sense of security and satisfaction. Staring at the "iron Lattice lattice every Duan lattice. The public security organs Service Development and protect the livelihood of the people, the most direct service is to create a good public security environment, let people have a sense of security, let investors to worry. If a burglary can prevent cases of multiple masses safety feeling low, the masses "iron", means that our work dereliction of duty. Therefore, to enhance the people a sense of security and satisfaction, so that "iron" more and less, is our biggest goal. As long as we earnestly to press incidence, enhance people's sense of security and satisfaction as the starting point and the foothold, will be able to work in the Party committee and government approval to obtain the support of the masses, the vast majority of the civilian police support, will not Continue to make new achievements. Second, we must find a good breakthrough point. In recent years, through to the primary research and development of visit, a sense of security satisfaction survey and public reception of visitors, accept masses to complain etc. channel, the police and the masses reflects the tension of police, theft cases, after the report did not deal with the results, the police slowly, service attitude stiff problems. Analysis the above problems, the breakthrough point is to solve tension of police and individual police service attitude problem. To solve the problems of shortage of police