【作者】 卢兴红；

【导师】 张兰桐；

【作者基本信息】 河北医科大学 ， 药物分析学， 2003， 硕士

【摘要】 栀子是茜草科植物栀子Gardenia jasminoides Ellis.的干燥成熟果实，中医学认为栀子具有护肝、利胆、降压、镇静、止血、消肿等作用，临床上常用于治疗黄疸型肝炎、扭挫伤、高血压、糖尿病等疾病。栀子苷是栀子中主要的有效成分，中国药典2000年版已把栀子苷确定为栀子原药材的质量控制指标。对于含栀子的复方制剂如加味逍遥丸和清开灵注射液，建立栀子苷的测定方法，可为制剂的质量控制提供一项指标。本研究以栀子全果、栀子皮、栀子仁、加味逍遥丸和清开灵注射液为研究对象，建立了同时测定栀子及含栀子制剂中栀子苷的方法，为中药栀子及其含栀子制剂的质量控制提供一种快速、简便的方法。目的：建立高效液相色谱法测定栀子各部位及含栀子制剂的栀子苷含量，为栀子及含栀子制剂的质量控制提供一项指标。方法：①提取：用超声波振荡提取样品中的栀子苷。②色谱条件：选用合适的色谱柱，调整流动相的组成、配比、流速，使栀子苷色谱峰的分离符合测定要求。③标准曲线的绘制：配制一系列浓度的栀子苷对照品溶液，测定峰面积；以栀子苷的含量为横坐标，相应的峰面积为纵坐<WP=5>标，制备标准曲线。④最低检测限的确定：将栀子苷对照品溶液逐步稀释，当信噪比S/N≥3时确定为最低检测限。⑤稳定性试验：取样品溶液分别放置8h后进样测定峰面积。⑥精密度试验：取同一份栀子药材样品溶液，重复进样6次，测定峰面积。⑦重复性试验：取栀子全果药材、加味逍遥丸和清开灵注射液，各平行制备同一浓度的样品5份。⑧回收率试验：取已知栀子苷含量的栀子药材及含栀子制剂，分别加入适量的栀子苷对照品，各平行制备同一浓度的样品5份。⑧样品测定： 在上述色谱条件下，测定栀子全果、栀子皮、栀子仁、加味逍遥丸和清开灵注射液中栀子苷的含量。结果：①提取：超声波振荡提取，具有快速、简便和稳定等优点。②色谱条件：以Kromasil C18为色谱柱，以乙腈-水（10:90，V/V）为流动相，流速为1.0ml·min-1，进样量为20μl，检测波长为238nm。在此色谱条件下，样品色谱峰的分离度大于1.5，理论塔板数大于6000，不对称因子0.98~1.04，阴性样品对栀子苷的测定无干扰。③栀子苷在0.596~2.980μg范围内，呈良好的线性关系，回归方程为Y=4.966X-0.0515，r=0.9998（n=5），④最低检测量为11.2ng。⑤样品在8h内稳定，其RSD分别为1.5%、0.75%和1.8%。⑥样品的精密度良好，其RSD值为0.37%。⑦样品的重复性好，栀子苷的含量分别为41.9mg·g-1、0.43 mg·g-1和30.4mg·100ml-1，其RSD分别为0.46%、1.3%和1.7%。⑧样品回收率分别为99.20%、99.85%和99.49%，其相应的RSD值为0.64%、0.68%和0.57%。⑨依据上述色谱条件进行分析，由标准曲线计算得到上述各药的栀子<WP=6>苷的含量，它们分别是：栀子全果41.8mg·g-1、栀子皮12.3mg·g-1、栀子仁67.3mg·g-1、加味逍遥丸4.26mg·g-1、清开灵注射液0.316mg·ml-1。结论：本文首次建立了用同一高效液相色谱法测定栀子及其制剂中栀子苷的含量，此方法专属性强、操作简便、结果准确、精密度和稳定性良好，可用于栀子及其制剂中栀子苷的含量测定。更多还原

【Abstract】 Gardenia Jasminoides Ellis, Rubia cordifolia plant’s dry mellow fruit, can reduce blood pressure, protect liver, be secretory bile, stanch and so on in Traditional Chinese Medicine theory and can relieve hypertension, diabetes, hepatitis in clinic. Geniposide, one of bioactive components, has been used to control the quality of Gardenia Jasminoides Ellis in Chinese Pharmacopoeia (2000 edtion, the first part). But determination of the content of geniposide in many preparations including Gardenia Jasminoides Ellis such as Jiaweixiaoyao pill and Qingkailing injection may give one guide line for their quality control. The paper studied the method with RP-HPLC to determine the geniposide in Gardenia Jasminoides Ellis fruit, peel, seed, Jiaweixiaoyao pill and Qingkailing injection, synchronously. It provides one quick, simple method to control quality of Gardenia Jasminoides Ellis and the preparations containing Gardenia Jasminoides Ellis.<WP=11>Objective：To establish a method for the assay of geniposide in Gardenia jasminoides Ellis and the preparations containing Gardenia jasminoides Ellis in order to provide one guide line for their quality control.Methods: ①Separate and extract geniposide from Gardenia jasminoides Ellis fruit, peel, seed, Jiaweixiaoyao pill and Qingkailing injection by supersonic wave-extraction.②Choose appropriate column, different formulation and proportion of mobile phase and adjust optimal flow rate and wavelength in order to separate geniposide well.③Preparation of standard curve: prepare a series of the reference solution, determine peak area，then the regression equation was obtained with the content of geniposide as abscissa, the relevant peak area as ordinate. ④Establishment of the lowest detection limitation: Dilute the reference solution until the value of S/N is more than 3. The relevant concentration is the lowest detection limitation. ⑤In the experiment of stability, transfer the sample solution from Gardenia jasminoides Ellis and the preparations containing Gardenia jasminoides Ellis to determine the relevant peak areas at 1, 2, 3, 4, 5, 6, 7 and 8 hour, respectively. ⑥In the experiment of precision, take the same test solution, inject it into the apparatus for six times, determine the peak areas of geniposide and calculate the concentrations of geniposide according to the same standard curve with the same apparatus in the same day, record the chromatograms, then<WP=12>calculate the precision. ⑦In the experiment of reproducibility, prepare one concentration sample solutions of Gardenia jasminoides Ellis and the preparations containing Gardenia jasminoides Ellis, repeat each concentration solution five times in the same way, respectively. ⑧In the experiment of recovery, transfer 5 shares of Gardenia Jasminoides Ellis and the preparations containing Gardenia jasminoides Ellis , respectively, add same volume of reference solution of geniposide. Inject them into the apparatus and determine the content of every sample. ⑨The samples of Gardenia Jasminoides Ellis fruit, peel, seed, Jiaweixiaoyao pill and Qingkailing injection were analyzed under above-mentioned conditions.Results: ①The method of supersonic wave-extraction was simple, quick and stable. ②The HPLC system was carried out on Kromasil C18 (4.6mm×250mm, 5μm) with acetonitrile-water (10:90) as a mobile phase, and the geniposide was detected at wavelength of 238 nm. Under the above circumstance, the peaks of these samples were separated well with the resolution of more than 1.5. Theoretical plates was more than 6000 and the tailing factor for the peak of geniposide was between 0.98 ~ 1.04. The result showed that the negative samples did not affect the determination of geniposide. ③The regression equation was: Y=4.9662X-0.0515, r=0.9998 (n=5), the linear range of geniposide was from 0.596μg to 2.980μg. ④The lowest<WP=13>detection limitation was 11.2ng. ⑤The samples were stable for 8 hours, and the relevant RSD values were 1.5%, 0.75% and 1.8%, respectively.⑥ The precision of sample was g更多还原