治疗慢性肾功能衰竭的海洋复方新药初步研究
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摘要
本论文在FPS研究的基础上,结合中医配伍理论,研究将FPS与几味传统中药-大黄、黄芪、泽泻、益母草的提取物组合成治疗慢性肾功能衰竭的海洋复方新药物。由于FPS研究相对成熟,故对复方药物中FPS外其它组分(以下简称复方)的提取、分离工艺以及制定复方质量的标准和检测方法等药学内容进行研究,为下一步新药的开发打下基础。
1.对复方的醇提与水提进行了比较,确定了复方最佳提取方法-醇提。采用正交实验设计方法对醇提工艺进行优化,以复方中主要有效成分的含量为指标进行综合评价,得到了复方提取的最佳工艺:8倍量95%乙醇回流提取两次,1.5h/次,并对该提取工艺进行了验证实验,结果表明本工艺提取物中各主要有效成分的含量均较高,是可靠可行的。
2.采用微波水提法,对复方中的单味药-黄芪的主要有效成分黄芪皂苷的提取进行了研究,通过正交实验设计方法优选出了最佳提取工艺为20倍量水、微波功率800 W、对60目黄芪提取两次、15 min/次。将该工艺与传统乙醇回流提取方法进行了对比,结果表明该工艺快速、经济、有较好的皂苷提取率,是黄芪皂苷提取的很好的方法。
3.进行缺FPS的褐藻糖含量测定阴性对照实验,结果阴性无干扰,褐藻糖含量测定可以作为控制复方质量的一个指标。分别做缺大黄和缺黄芪阴性对照实验,结果总蒽醌和总皂苷含量测定阴性干扰较大,大黄素含量测定阴性无干扰,确定大黄素含量测定为控制复方质量的另一指标。
4.进行复方提取物质量控制指标之一-大黄素含量测定的分析方法学研究,找到了一稳定可行的方法-HPLC方法,该方法测定条件为:流动相甲醇与0.1%磷酸比例为85:15,柱温40℃,通过ODS柱分离,用二极管阵列检测器在436 nm下测定含量。该方法测定复方提取物中大黄素含量精密度高、专一性好、干扰小,可用来控制复方的质量。
Based on the study of FPS and the coupling theory of traditional Chinese medicine, Fufang New Marine Drug was composed of extract from some traditional Chinese medicine and FPS. The treat of Fufang New Marine Drug for chronic renal failure was studied in this thesis. Because of relatively intact study on FPS, the other part of Fufang New Marine Drug, which was named as Fufang in this thesis, was studied, including extraction, isolation, establishment of quality control standard, etc. This study was the base for the exploitation of new drug. The results are listed as following:
1. Ethanol extraction was compared with water extraction and ethanol extraction was confirmed as the better extraction technology of Fufang. Then ethanol extraction technology was optimized by orthogonal test design method, with content of effective components extracted from Fufang as evaluating standards, the optimum extraction technology was the following: Fufang was extracted twice with 8 fold-volume 95% ethanol, 1.5h for each time.
2. Radix Astragali, a composition of Fufang, was extracted by microwave water-extraction. Then comparison was made between microwave water-extraction and traditional ethanol extraction. Microwave water-extraction was confirmed as an economical, easy and effective method. An optimum extraction technology was obtained by orthogonal design method: 60-mash powder of Radix Astragali was extracted twice with 20 fold-volume water and 800 W microwave power, 15 min for each time.
3. The content of fucoidan in Fufang New Marine Drug was compared with that in Fufang. Negative extract had no influence on the determination of fucoidan. So determination of fucoidan was chosen as a standard for the quality control ofr Fufang New Marine Drug. Fufang without rhubarb and Fufang without Radix Astragali were prepared respectively. Then the content of emodin and total anthraquinones in Fufang without rhubarb was compared with that in Fufang. The content of saponins in Fufang without Radix Astragali was compared with that in Fufang. The determination of emodin in Fufang was confirmed as another standard for quality control of Fufang New Marine Drug.
4. Determination method of emodin in Fufang was studied. HPLC method, an accurate, steady, highly specific method, was established as the determination method for quality control of Fufang New Marine Drug.
1.对复方的醇提与水提进行了比较,确定了复方最佳提取方法-醇提。采用正交实验设计方法对醇提工艺进行优化,以复方中主要有效成分的含量为指标进行综合评价,得到了复方提取的最佳工艺:8倍量95%乙醇回流提取两次,1.5h/次,并对该提取工艺进行了验证实验,结果表明本工艺提取物中各主要有效成分的含量均较高,是可靠可行的。
2.采用微波水提法,对复方中的单味药-黄芪的主要有效成分黄芪皂苷的提取进行了研究,通过正交实验设计方法优选出了最佳提取工艺为20倍量水、微波功率800 W、对60目黄芪提取两次、15 min/次。将该工艺与传统乙醇回流提取方法进行了对比,结果表明该工艺快速、经济、有较好的皂苷提取率,是黄芪皂苷提取的很好的方法。
3.进行缺FPS的褐藻糖含量测定阴性对照实验,结果阴性无干扰,褐藻糖含量测定可以作为控制复方质量的一个指标。分别做缺大黄和缺黄芪阴性对照实验,结果总蒽醌和总皂苷含量测定阴性干扰较大,大黄素含量测定阴性无干扰,确定大黄素含量测定为控制复方质量的另一指标。
4.进行复方提取物质量控制指标之一-大黄素含量测定的分析方法学研究,找到了一稳定可行的方法-HPLC方法,该方法测定条件为:流动相甲醇与0.1%磷酸比例为85:15,柱温40℃,通过ODS柱分离,用二极管阵列检测器在436 nm下测定含量。该方法测定复方提取物中大黄素含量精密度高、专一性好、干扰小,可用来控制复方的质量。
Based on the study of FPS and the coupling theory of traditional Chinese medicine, Fufang New Marine Drug was composed of extract from some traditional Chinese medicine and FPS. The treat of Fufang New Marine Drug for chronic renal failure was studied in this thesis. Because of relatively intact study on FPS, the other part of Fufang New Marine Drug, which was named as Fufang in this thesis, was studied, including extraction, isolation, establishment of quality control standard, etc. This study was the base for the exploitation of new drug. The results are listed as following:
1. Ethanol extraction was compared with water extraction and ethanol extraction was confirmed as the better extraction technology of Fufang. Then ethanol extraction technology was optimized by orthogonal test design method, with content of effective components extracted from Fufang as evaluating standards, the optimum extraction technology was the following: Fufang was extracted twice with 8 fold-volume 95% ethanol, 1.5h for each time.
2. Radix Astragali, a composition of Fufang, was extracted by microwave water-extraction. Then comparison was made between microwave water-extraction and traditional ethanol extraction. Microwave water-extraction was confirmed as an economical, easy and effective method. An optimum extraction technology was obtained by orthogonal design method: 60-mash powder of Radix Astragali was extracted twice with 20 fold-volume water and 800 W microwave power, 15 min for each time.
3. The content of fucoidan in Fufang New Marine Drug was compared with that in Fufang. Negative extract had no influence on the determination of fucoidan. So determination of fucoidan was chosen as a standard for the quality control ofr Fufang New Marine Drug. Fufang without rhubarb and Fufang without Radix Astragali were prepared respectively. Then the content of emodin and total anthraquinones in Fufang without rhubarb was compared with that in Fufang. The content of saponins in Fufang without Radix Astragali was compared with that in Fufang. The determination of emodin in Fufang was confirmed as another standard for quality control of Fufang New Marine Drug.
4. Determination method of emodin in Fufang was studied. HPLC method, an accurate, steady, highly specific method, was established as the determination method for quality control of Fufang New Marine Drug.
引文
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[10] 焦淑芳,喻红.温阳通腑降浊法治疗慢性肾衰竭阳虚浊毒证疗效观察[J].湖南中医学院学报,2001,21(2):51-52.
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[12] 张春艳,王建明,张佩青.参芪地黄冲剂对慢性肾衰血流动力学的影响[J].现代中西医结合杂志,2001,lO(23):2229-2230.
[13] 张小娟,余俊文.肾维宁冲剂对慢性肾功能衰竭患者尿微量蛋白及脂质代谢疗效观察[J].新中医,2000,32(7):33-34.
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