藏药红景天颗粒中多指标组分的 体外 溶出度测定
Summary
在这里，我们在 体外测试了红景天颗粒（RG）的溶解度，绘制了红景天苷，没食子酸和没食子酸乙酯在超纯水中的溶解曲线，并将曲线拟合到不同的数学模型中。该协议为RG的 体内 生物等效性和体内外体 外 相关性研究提供了信息和指导。
Abstract
藏药红景天颗粒（RG）成分复杂，RG的整体质量难以确定。因此，建立一种测定RG多组分 体外 溶出度的方法对质量控制具有重要意义。本研究采用中国药典（2020年版）第四通则0931的第二桨法，符合美国药典（USP）的器具2。将溶解装置设置为100 rpm的转速，以超纯水为溶解介质。在每个时间点收集1 mL的样品体积。采用高效液相色谱法测定没食子酸、红景天苷和没食子酸乙酯在不同时间点的累积溶出量。最后，绘制了溶解曲线，并将曲线拟合到GompertzMod，Gompertz，Logistic和Weibull方程。结果表明：没食子酸在RG中的累积溶出度在1 min时大于80%，红景天苷和没食子酸乙基在5 min时的累积溶出度超过65%，30 min后各指标组分的累积溶出度下降。曲线拟合表明，GompertzMod方程是RG每个指数分量的最佳拟合模型。总之，本协议中描述的溶出度测试方法简单，准确且可靠。它可以表征RG中指标组分的体 外溶出行为，为RG的质量控制和其他民族化合物的质量评价提供方法学参考。
Introduction
在中国，心血管疾病患病率持续上升，心血管疾病发病率和死亡率居中国居民首位1。冠心病的心绞痛是由冠状动脉粥样硬化引起的管腔狭窄引起的，导致冠状动脉血液供应相对不足和心肌缺血缺氧2。近年来，中医药治疗冠心病的疗效得到了众多医生的认可3。
中医药在缓解临床症状和提高患者生活质量方面发挥着重要作用4.红景天颗粒（RG）是从青藏高原药用植物红 景天L 中提取和精制的。RG的主要成分是红景天苷、红景天和类黄酮5，6。RG具有补气7 ，活化和促进血液循环缓解疼痛的作用。临床上用于治疗气虚血瘀引起的胸梗阻、冠心病、心绞痛8.仅含量测定并不能完全反映药物的内在质量，因为 体外崩 解和溶出都会影响药物的生物利用度和功效9，10。中药溶出的检验方法有旋转篮法、桨法、小杯法等。旋转篮法的缺点是旋转过程中只有旋转篮的外部与溶解介质接触，这并不能反映现实世界的溶解行为。桨法可以克服上述缺点，使其比篮子法更适合一些固体中药制剂11。目前尚无RG体 外 溶出分析的报道。为了更全面地控制RG的质量，研究了RG中3种指标成分（没食子酸、红景天苷和没食子酸乙酯）的溶出行为。本研究为RG的质量控制提供了数据，为其他民族化合物制剂的质量评价提供了方法学参考。
Protocol
1. 溶液制备 制备对照物质储备溶液：在电子分析天平上分别称取10.6 mg红景天苷、5.24 mg没食子酸和5.21 mg没食子酸乙酯，并将它们单独加入5 mL容量瓶中。然后，加入HPLC级甲醇溶解并稀释至5 mL。最后摇匀，得到质量浓度分别为2.120 mg/mL、1.048 mg/mL和1.042 mg/mL的对照品储备溶液。注意：对照物质储备溶液含有2.120 mg / mL红景天苷，1.048 mg / mL没食子酸和1.042mg / mL没食子酸乙酯作为随…
Representative Results
本研究显示，RG的精密度、稳定性、重复性和样品回收率均在中国药典（2020年第4卷）12规定的方法学范围内，表明该方法是可行的。经过反复调试，确定本研究中使用的洗脱梯度对RG中的三种指标组分具有良好的分辨率（图1）。RG中的3种指标组分在特定浓度范围内具有良好的线性关系（表2）。精密度测试结果（表3）显示，红景天?…
Discussion
溶出度试验是模拟固体口服制剂在胃肠道15中崩解和溶出的理想体外方法。是评价和控制固体口服制剂质量的重要指标。因此，溶出度试验在固体药物口服制剂的开发中起着至关重要的作用16。特别是随着中药（TCM）质量控制技术的发展，溶出度测定已逐步应用于中药和民族药复方制剂的筛选研究17，18。
<p c…Declarações
The authors have nothing to disclose.
Acknowledgements
这项工作由国家重点研发计划（2017YFC1703904）、成都中医药大学-企业（西藏红景天制药控股有限公司）合作项目（1052022040101）;四川省科技厅区域创新合作项目（2020YFQ0032）;青海省科技厅重点研发转化计划（2020-SF-C33）。
Materials
| Chromatographic column | ZORBAX Eclipse | XDB-C18 | 4.6 mm x 250 mm, 5 µm |
| Drug dissolution tester | Shanghai Huanghai Pharmaceutical Inspection Instrument Co., Ltd. | RCZ-6B3 | |
| Electronic analytical balance | Shanghai Liangping Instruments Co., Ltd. | FA1004 | |
| Ethyl gallate (HPLC, ≥98%) | Chengdu Desite Biotechnology Co., Ltd. | DSTDM006301 | |
| Function drawing software | OriginLab Corporation, Northampton, MA, USA | 2022 | |
| Gallic acid (HPLC, ≥98%) | Chengdu Desite Biotechnology Co., Ltd. | DSTDM000802 | |
| High performance liquid chromatography | Agilent Technologies Singapore (International) Pte. Ltd. | Agilent 1260 Infinity  |
|
| HPLC grade methanol | Thermo Fisher Scientific (China) Co., Ltd. | 216565 | |
| Injector | Chengdu Xinjin Shifeng Medical Apparatus & Instrument Co., Ltd. | 0.7 (22 G) | |
| Millipore filter | Tianjin Jinteng Experimental Equipment Co., Ltd | φ13 0.22 Nylon66 | |
| Rhodiola granules | Tibet Nodikang Pharmaceutical Co., Ltd. | 210501 | |
| Salidroside (HPLC, ≥98%) | Chengdu Desite Biotechnology Co., Ltd. | DST200425-037 | |
| Ultra pure water systemic | Merck Millipore Ltd. | Milli-Q | |
| Ultrasonic cleansing machine | Ningbo Xinyi Ultrasonic Equipment Co., Ltd | SB-8200 DTS |
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Citar este artigo
Du, Q., He, Q., Zhang, F., Mi, J., Li, Y., Wang, S., Zhang, Y. An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules. J. Vis. Exp. (189), e64670, doi:10.3791/64670 (2022).