保留灌肠剂—复方黄芪肠宁颗粒的药学部分研究
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摘要
目的:研制中药复方制剂-复方黄芪肠宁颗粒;建立其质量标准;考察其初步稳定性。
方法:采用小鼠实验性溃疡性结肠炎模型,以模型小鼠的体征、结肠溃疡点数和病理切片中杯状细胞和炎症淋巴细胞的变化情况作为指标,选择处方的提取条件;
以人参皂苷Rg_1、人参皂苷Rb_1、三七皂苷R_1的含量之和为指标,采用正交实验设计,优选三七药材的提取工艺;
以黄芪甲苷、总黄酮以及总固体得率为指标,采用正交实验设计,优选黄芪等群药的水提工艺;
以黄芪甲苷、总黄酮以及总固体得率为指标,考察黄芪等群药水提液的精制工艺;
以黄芪甲苷、总黄酮以及PNS含量为指标,对水提液、醇提液的浓缩、干燥工艺进行考察;
通过浸膏粉吸湿性试验,以改善吸湿率为指标,对制粒的辅料种类和用量进行筛选;
采用湿法制粒法制备颗粒;
在质量标准研究中,采用薄层色谱法对制剂中的黄芪、三七、补骨脂、地榆进行鉴别;采用HPLC-ELSD法测定制剂中的黄芪甲苷含量;采用HPLC法测定制剂中PNS的含量;
通过加速试验和长期试验法考察了本品的初步稳定性。
结果:确定三七采用单独提取的方法,优选的提取工艺为加70%乙醇6倍量,提取3次,每次0.5h;
黄芪等其余药材采用混合提取的方法,优选的提取工艺为加水9倍量,提取3次,每次1.0h;
采用乙醇沉淀法对黄芪等群药的水提液进行精制,条件是将药液浓缩至相对密度1.05(60℃)、调整醇沉浓度为50%,后静置24h,滤过,回收乙醇;
药液在温度70℃,真空度为0.08～0.1Mpa条件下浓缩,浓缩到相对密度是1.10(60℃)的浸膏并置减压干燥器中干燥,条件为温度70℃,真空度为0.08～0.1Mpa;
以干浸膏量70%的乳糖作为成型辅料,湿法制粒所得颗粒棕色,颗粒的堆密度是0.53g/ml。休止角是35.1度,临界相对湿度是62%。粒度、溶化性、水份均符合2005年版《中国药典》一部附录IC颗粒剂项下规定;
质量标准研究中,黄芪、三七、补骨脂、地榆薄层色谱斑点清晰,分离度好,无阴性干扰。黄芪甲苷在1.430μg～14.30μg范围内峰面积对数值与进样量对数值呈良好的线性关系,平均回收率是100.6%,RSD=1.1%,初步确定复方黄芪肠宁颗粒中含黄芪甲苷的量为不少于0.15mg/g;人参皂苷Rg_1、人参皂苷Rb_1、三七皂苷R_1分别在1.285μg～12.85μg、1.193μg～11.93μg、0.6220μg～6.216μg范围内进样量与峰面积呈良好的线性关系,PNS平均回收率是99.6%,RSD=1.9%,初步确定复方黄芪肠宁颗粒中含三七以PNS计不少于6.5776mg/g。
选择复合铝箔膜作为包装材料,铝袋材质为PET/Al/PE。在密封状态下保存。本品每袋装15g,含生药量71.5g,使用时加开水40ml溶解,降至体温后供保留灌肠用,一日2次。
经过加速试验和长期试验考察,本品性状、鉴别、水分、粒度、溶化性及含量测定未发生明显变化。
结论:复方黄芪肠宁颗粒制备工艺合理可行;质量控制方法简单、专属性强、重现性好、回收率高,可有效控制产品质量;本制剂质量稳定。
Objective: To study the preparation precedure of compound huangqi changning granules, establish the quality standard and investigate the preliminary stability.
Methods: The preliminary extraction technology was chosen by the indexes of the changes of physical sign, the numbers of colonic ulcer, caliciform cells, and inflammation lymphocyte by the application of the experimental ulcerative colitis animal model of rat. Orthogonal test was employed for selecting the optimum of extraction technology by the indexes of the contents of ginsenoside Rg_1, ginsenoside Rb_1, notoginsenoside R_1 of the herbs of Radix Notoginseng. Orthogonal test was also employed for selecting the optimal extraction technology of Radix astragali and other herbs with colligation score by the indexes of Astragaloside IV, total solid matters and total flavone in water extraction. And they were also used to investigate the purification technology of water extraction. The concentrated and dried technology of water extraction and ethanol extraction by the contents of Astragaloside IV, total flavone and Panax Notoginseng Saponins. The kind and dosage of adjuvant were investigated by the index of improved hygroscopicity with the experiment of hygroscopicity of powdered extract. The granules were prepared by the method of wet granulation. Radix Astragali, Radix Notoginseng, Radix Sanguisorbae and Fructus Psoraleae in compound huangqi changning granules were identified by TLC; The content of Astragaloside IV in this preparation was determined by HPLC-ELSD in the study of quality standard. And the content of Panax Notoginseng Saponins in this preparation was determined by HPLC. Preliminary stability of the preparation was investigated by accelerated test and long-term testing. Results: Six volumes of 70% ethanol, extracting 3 times with 0.5 hour for each time was considered the optimum extraction technology of the herbs of Radix Notoginseng. And nine volumes of water, extracting 3 times with one hour for each time was considered the optimum extraction technology of Radix astragali and other herbs. Water extraction was purified by ethanol precipitation, and it was condensed to the relative density of 1.05 under the condition of 60℃, and the content of ethanol was adapted to 50%, standed by 24 hours, filtrated. The water extration and ethanol extraction were concentrated and then vacuum dried under the condition of 70℃, vacuum=0.08～0.1Mpa. 70% weight of dry powered extraction was applied to prepare the granules. And the granules were brown. The bulk density was 0.53g/ml. The angle of repose was 35.1 degree. The critical relative humidity was 62%. Granularity, moisture content, and solubility were qualified all the rules of granule in《Chinese Pharmacopoeia》2005ed Vol I. The quality identification by TLC was specific,having a good separation and a clear spot in each thin layer plate. Astragaloside IV was linear with the logarithm of the peak area in the range of 1.430μg～14.30μg, and the average recovery was 100.6%, RSD=1.1%(n=6). The content of Astragaloside IV was no less than 0.15mg/g. Ginsenoside Rg_1, ginsenoside Rb_1 and notoginsenoside R_1 were linear with the peak area in the range of 1.285μg～12.85μg, 1.193μg～11.93μg, 0.6220μg～6.216μg respectively, and the average recovery of Panax Notoginseng Saponins was 99.6%, RSD=1.9%(n=6). The content of Panax Notoginseng Saponins was no less than 6.5776mg/g. Aluminium foil was chosen to be packing material, and it was PET/Al/PE. The granules were seal up to preserve. Every bag was filled to 15g. It was dissolved by using 40ml of boiled water and retation enema when it was lowered to body temperature for two times a day. The charater, identification, moisture content, solubility, granularity, assaying of Astragaloside IV hadn’t been changed obviously by accelerated test and long-term testing.
Conclusion: The preparation precedure of compound huangqi changning granules was reasonable, practical. The evaluation methods are simple, specificity, reproducible and can be used in the quality control of compound huangqi changning granules. And the quality of the sample was steady as well.
方法:采用小鼠实验性溃疡性结肠炎模型,以模型小鼠的体征、结肠溃疡点数和病理切片中杯状细胞和炎症淋巴细胞的变化情况作为指标,选择处方的提取条件;
以人参皂苷Rg_1、人参皂苷Rb_1、三七皂苷R_1的含量之和为指标,采用正交实验设计,优选三七药材的提取工艺;
以黄芪甲苷、总黄酮以及总固体得率为指标,采用正交实验设计,优选黄芪等群药的水提工艺;
以黄芪甲苷、总黄酮以及总固体得率为指标,考察黄芪等群药水提液的精制工艺;
以黄芪甲苷、总黄酮以及PNS含量为指标,对水提液、醇提液的浓缩、干燥工艺进行考察;
通过浸膏粉吸湿性试验,以改善吸湿率为指标,对制粒的辅料种类和用量进行筛选;
采用湿法制粒法制备颗粒;
在质量标准研究中,采用薄层色谱法对制剂中的黄芪、三七、补骨脂、地榆进行鉴别;采用HPLC-ELSD法测定制剂中的黄芪甲苷含量;采用HPLC法测定制剂中PNS的含量;
通过加速试验和长期试验法考察了本品的初步稳定性。
结果:确定三七采用单独提取的方法,优选的提取工艺为加70%乙醇6倍量,提取3次,每次0.5h;
黄芪等其余药材采用混合提取的方法,优选的提取工艺为加水9倍量,提取3次,每次1.0h;
采用乙醇沉淀法对黄芪等群药的水提液进行精制,条件是将药液浓缩至相对密度1.05(60℃)、调整醇沉浓度为50%,后静置24h,滤过,回收乙醇;
药液在温度70℃,真空度为0.08～0.1Mpa条件下浓缩,浓缩到相对密度是1.10(60℃)的浸膏并置减压干燥器中干燥,条件为温度70℃,真空度为0.08～0.1Mpa;
以干浸膏量70%的乳糖作为成型辅料,湿法制粒所得颗粒棕色,颗粒的堆密度是0.53g/ml。休止角是35.1度,临界相对湿度是62%。粒度、溶化性、水份均符合2005年版《中国药典》一部附录IC颗粒剂项下规定;
质量标准研究中,黄芪、三七、补骨脂、地榆薄层色谱斑点清晰,分离度好,无阴性干扰。黄芪甲苷在1.430μg～14.30μg范围内峰面积对数值与进样量对数值呈良好的线性关系,平均回收率是100.6%,RSD=1.1%,初步确定复方黄芪肠宁颗粒中含黄芪甲苷的量为不少于0.15mg/g;人参皂苷Rg_1、人参皂苷Rb_1、三七皂苷R_1分别在1.285μg～12.85μg、1.193μg～11.93μg、0.6220μg～6.216μg范围内进样量与峰面积呈良好的线性关系,PNS平均回收率是99.6%,RSD=1.9%,初步确定复方黄芪肠宁颗粒中含三七以PNS计不少于6.5776mg/g。
选择复合铝箔膜作为包装材料,铝袋材质为PET/Al/PE。在密封状态下保存。本品每袋装15g,含生药量71.5g,使用时加开水40ml溶解,降至体温后供保留灌肠用,一日2次。
经过加速试验和长期试验考察,本品性状、鉴别、水分、粒度、溶化性及含量测定未发生明显变化。
结论:复方黄芪肠宁颗粒制备工艺合理可行;质量控制方法简单、专属性强、重现性好、回收率高,可有效控制产品质量;本制剂质量稳定。
Objective: To study the preparation precedure of compound huangqi changning granules, establish the quality standard and investigate the preliminary stability.
Methods: The preliminary extraction technology was chosen by the indexes of the changes of physical sign, the numbers of colonic ulcer, caliciform cells, and inflammation lymphocyte by the application of the experimental ulcerative colitis animal model of rat. Orthogonal test was employed for selecting the optimum of extraction technology by the indexes of the contents of ginsenoside Rg_1, ginsenoside Rb_1, notoginsenoside R_1 of the herbs of Radix Notoginseng. Orthogonal test was also employed for selecting the optimal extraction technology of Radix astragali and other herbs with colligation score by the indexes of Astragaloside IV, total solid matters and total flavone in water extraction. And they were also used to investigate the purification technology of water extraction. The concentrated and dried technology of water extraction and ethanol extraction by the contents of Astragaloside IV, total flavone and Panax Notoginseng Saponins. The kind and dosage of adjuvant were investigated by the index of improved hygroscopicity with the experiment of hygroscopicity of powdered extract. The granules were prepared by the method of wet granulation. Radix Astragali, Radix Notoginseng, Radix Sanguisorbae and Fructus Psoraleae in compound huangqi changning granules were identified by TLC; The content of Astragaloside IV in this preparation was determined by HPLC-ELSD in the study of quality standard. And the content of Panax Notoginseng Saponins in this preparation was determined by HPLC. Preliminary stability of the preparation was investigated by accelerated test and long-term testing. Results: Six volumes of 70% ethanol, extracting 3 times with 0.5 hour for each time was considered the optimum extraction technology of the herbs of Radix Notoginseng. And nine volumes of water, extracting 3 times with one hour for each time was considered the optimum extraction technology of Radix astragali and other herbs. Water extraction was purified by ethanol precipitation, and it was condensed to the relative density of 1.05 under the condition of 60℃, and the content of ethanol was adapted to 50%, standed by 24 hours, filtrated. The water extration and ethanol extraction were concentrated and then vacuum dried under the condition of 70℃, vacuum=0.08～0.1Mpa. 70% weight of dry powered extraction was applied to prepare the granules. And the granules were brown. The bulk density was 0.53g/ml. The angle of repose was 35.1 degree. The critical relative humidity was 62%. Granularity, moisture content, and solubility were qualified all the rules of granule in《Chinese Pharmacopoeia》2005ed Vol I. The quality identification by TLC was specific,having a good separation and a clear spot in each thin layer plate. Astragaloside IV was linear with the logarithm of the peak area in the range of 1.430μg～14.30μg, and the average recovery was 100.6%, RSD=1.1%(n=6). The content of Astragaloside IV was no less than 0.15mg/g. Ginsenoside Rg_1, ginsenoside Rb_1 and notoginsenoside R_1 were linear with the peak area in the range of 1.285μg～12.85μg, 1.193μg～11.93μg, 0.6220μg～6.216μg respectively, and the average recovery of Panax Notoginseng Saponins was 99.6%, RSD=1.9%(n=6). The content of Panax Notoginseng Saponins was no less than 6.5776mg/g. Aluminium foil was chosen to be packing material, and it was PET/Al/PE. The granules were seal up to preserve. Every bag was filled to 15g. It was dissolved by using 40ml of boiled water and retation enema when it was lowered to body temperature for two times a day. The charater, identification, moisture content, solubility, granularity, assaying of Astragaloside IV hadn’t been changed obviously by accelerated test and long-term testing.
Conclusion: The preparation precedure of compound huangqi changning granules was reasonable, practical. The evaluation methods are simple, specificity, reproducible and can be used in the quality control of compound huangqi changning granules. And the quality of the sample was steady as well.
引文
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