【作者】 王芳；

【导师】 张开莲；

【作者基本信息】 泸州医学院 ， 药理学， 2009， 硕士

【摘要】 目的:采用气相色谱法对积雪草挥发油化学成分指纹图谱进行研究,建立积雪草挥发油的气相指纹图谱标准,用于积雪草药材挥发油的质量控制,并为雪抑乐质量标准的制订提供依据。方法:采用水蒸气蒸馏法提取积雪草挥发油。利用气相色谱法测定积雪草挥发油指纹图谱,通过考察不同条件下的分离效果,优选色谱条件:色谱柱的选择,柱温或程序升温模式,载气流速,进样量;考察供试品溶剂对测定的干扰;并进一步考察指纹图谱的稳定性、精密度、重现性,最终确定积雪草指纹图谱分析的气相色谱条件。按所确定的色谱条件和测定方法,对积雪草所含挥发油成分的气相色谱指纹图谱进行研究。先后对10批积雪草药材供试品进行GC检测分析,以石竹烯为参照物,按“中药注射剂指纹图谱研究的技术要求”计算相对保留时间及峰面积比值,标定共有指纹峰,并对10批药材进行相似度比较。结果:优选出的色谱条件为:色谱柱:HP-5Ms石英毛细管柱(30m×0.25mm);柱温:初温90℃(保持5min)以10℃/min升至170℃(保持8min)以5℃/min升至250℃(保持2min);载气:高纯氮气,流量1.2ml/min(0.12Mpa);补充气:氮气,流量40ml/min;分流比:1:10;燃烧气:氢气,流量40ml/min;助燃气;空气,流量500ml/min;检测器:氢火焰离子化检测器(FID),检测器温度270℃,气化室温度235℃;进样量:1ul。供试品溶剂以选择石油醚(30～60℃)对测定无干扰。方法学考察结果:精密度试验共有峰的相对保留时间、相对峰面积的RSD为0～2.78%,重现性试验共有峰相对峰面积、相对保留时间的RSD为0.02%～2.81%,供试品在8小时之内检测稳定,共有峰的相对保留时间、相对峰面积的RSD为0.01%～2.80%。对10批积雪草药材供试品进行GC检测分析,积雪草GC指纹图谱中共有32个共有峰,共有峰峰面积总百分含量为91.62±9.12。10批药材的GC指纹图谱相似度分别为0.904～0.965。结论:本研究所建立的GC分析方法可使积雪草挥发油中各组分达到较好分离效果,精密度、重现性、稳定性良好。10批药材的GC指纹图谱相似性高,所建立的GC指纹图谱标准符合国家指纹图谱技术要求,可用于积雪草药材挥发油的质量控制,并为雪抑乐质量标准的制订提供依据。更多还原

【Abstract】 Objective: By GC to carry out the research into the Fingerprint of the volatile oil in Centella Asiatica(L.) Urb., and to establish the standard of the volatile oil by GC Fingerprint. Method: The volatile oil in Centella asatica (L.) Urb. was extracted by wet distillation. By GC, the Fingerprint of the volatile oil in Centella asatica (L.) Urb. was determined. The separate effects under different conditions were studied so as to optimize the chromatographic condition, including the chromatographic column, the column temperature or the programmed heating mode, the flow rate of the carrier gas and the sample size. The intervention of the sample was also studied, as well as the stability, precision, reproducibility of the Fingerprint, which enabled the determination of the final chromatographic condition of the Fingerprint to Centella Asiatica(L.) Urb.. According to the optimized chromatographic condition and assay method, the Fingerprint of the volatile oil in Centella Asiatica(L.) Urb. was studied. Taking caryophyllene as the reference material, 10 batches of Centella Asiatica(L.) Urb.was assayed by GC, and the relative retention times and proportionalities of the peak area was calculated, the common peaks were marked and compared among the 10 according to the Technical Requirements for the Fingerprint Research into Chinese Crude Drug. Results: the optimal conditions were as follows: chromatographic column: HP-5Ms quartz capillary column (30mx0.25mm); column temperature: 90℃for the first 5 min, up to 170℃at the rate of 10℃/min, for 8 min, then up to 250℃at the rate of 5℃/min, for 2 min; the carrier gas: high pure nitrogen, flow rate 1.2ml/min (0.12Mpa); supplementary gas: nitrogen, flow rate 40ml/min; split ratio: 1:10; combustion air: hydrogen, flow rate 40ml/min; combustion-supporting gas: air, flow rate 500ml/min; detector: FID, detector temperature 270℃, vaporizer temperature 235℃; sample size: 1ul. Petroleum ether (30～60℃) was chosen as solvent. Results to the methodology study: the RSD of the relative retention time and the area of common peaks was 0～2.78% in the precision test, 0.02%-2.81% in the reproducibility test; 0.01%-2.80% in the stability test, which signified that the sample stayed stable within 8 hours. The Fingerprint of the 10 bathes of Centella asatica (L.) Urb. showed that there were 32 common peaks in the chromatogram, and the added area of which covered 91.62±9.12 in total percentage. Conclusion: The GC analytical method establish in this study can finely separate the ingredients in the volatile oil in Centella asatica (L.) Urb., and the precision, reproducibility and stability were satisfying. 10 batches of Centella asatica (L.) Urb. showed high similarity, and the established standard for the GC Fingerprint can meet the requirement from the guidance, thus can be applied to control the quality of Centella asatica (L.) Urb., meanwhile to provide a basis for the standard of the quality of Xue Yile.更多还原