固相微萃取与气相色谱-质谱联用分析蛇莓、贝母和卵巢癌的挥发性组分
文献类型：学位论文
| 作者 | 王晨旭 |
| 学位类别 | 博士 |
| 答辩日期 | 2015-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 中国科学院长春应用化学研究所 |
| 导师 | 徐经伟 |
| 关键词 | 固相微萃取 气相色谱-质谱联用 挥发性组分 蛇莓 贝母 卵巢癌 |
| 中文摘要 | 固相微萃取是一种较新的提取方法，与索氏提取、超声提取、水蒸气蒸馏等传统提取方法相比，固相微萃取在样品处理过程中不使用有机溶剂，绿色环保。此外，由于能够将提取、富集、进样集于一步完成，极大地简化了实验步骤，降低了操作过程中引入干扰与污染的机会。其与气相色谱-质谱联用可对挥发性组分进行快速分析。 中药蛇莓（Duchesnea indica (Andr.) Focke）属于蔷薇科蛇莓属，是多年生草本植物蛇莓的全草，又名蛇泡草、龙吐珠。蛇莓具有较强的抗氧化、抗菌消炎和中枢神经系统抑制作用。民间主要用于治疗发热、湿疹、带状疱疹和蛇虫咬伤等。近年来，由于发现蛇莓对多种肿瘤细胞的杀伤作用显著，其化学成分成为人们关注的焦点，已见报道的化合物种类主要包括萜类、酚酸类、黄酮类等。除了这些组分外，蛇莓疗效也可能与其挥发性组分相关，然而目前仅有一篇文献报道采用水蒸气蒸馏和气相色谱-质谱联用的方法分析印度产蛇莓。针对以上情况，本论文第二章中采用索氏提取、超声提取、水蒸气蒸馏和固相微萃取四种方法对国产蛇莓的挥发性组分进行了系统的研究。实验共鉴定出66种化合物，其中47种为首次在蛇莓中发现，这些新化合物的发现为蛇莓的开发利用提供重要参考。 贝母是百合科（Liliaceae）贝母属(Fritillaria)植物的干燥鳞茎，具有十分悠久的药用历史，始载于《神农本草经》，列为中品“气味辛、平，无毒，主伤寒烦热。淋沥邪气，病症，喉痹，乳难”。贝母种类繁多，仅《中国药典》一部中收录的药用贝母种类就包括川贝母、浙贝母、平贝母、伊贝母和湖北贝母，其中川贝母止咳化痰疗效显著，中药处方用量非常大。然而川贝母产量低，价格高，这导致市场中出现使用其他种类贝母冒充川贝母进行销售的现象。由于不同种类贝母的产地具有明显的区域性，我们推测生长环境的不同可能引起贝母的挥发性组分存在差异，能否通过挥发性组分对贝母种类进行区分引发了我们的研究兴趣。在本论文第三章中，我们采用固相微萃取与气相色谱-质谱联用技术对不同种类贝母的挥发性组分进行了鉴定，采用主成分分析与聚类分析对贝母的亲疏远近关系进行了分析。实验结果表明，相同种类的贝母在挥发性组分上具有更多的相似性，固相微萃取与气相色谱-质谱联用技术可以考虑作为区分贝母种类的辅助手段。 卵巢肿瘤有良性和恶性之分。成熟型畸胎瘤是一种典型的卵巢良性肿瘤，可通过手术治愈，致死率低。卵巢癌是卵巢恶性肿瘤，全球每年约有125000名女性死于卵巢癌，致死率居妇科肿瘤之首。缺乏有效的生物标志物是造成卵巢癌高致死率的主要原因。近年来，人们开始通过检测血液中挥发性化合物来寻找癌症生物标志物。在本论文第四章中，我们将固相微萃取与气相色谱-质谱联用法应用于卵巢癌、成熟型畸胎瘤和正常血样的分析，对影响提取效果的实验条件进行了优化。采用偏最小二乘判别分析法和独立样本t检验对实验数据进行处理，发现三组血液样本能够得到明显区分，在检出的28种挥发性组分中，8种化合物存在显著性差异，对这些化合物作为卵巢癌、成熟型畸胎瘤生物标志物的可能性进行了探讨。 |
| 英文摘要 | The newly developed solid-phase microextraction (SPME) is a solvent-free and environmentally sound technique, compared with the conventional extraction methods such as soxhlet extraction, ultrasonic extraction and steam distillation etc. Besides, SPME only needs one step to accomplish extraction and pre-concentration of analytes from sample matrix, which could simplify experimental procedure and avoid sample contamination. Solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME/GC-MS) is an effective method in analysing the volatile components of various samples. Belonging to the family Rosaceae, Duchesnea indica (Andr.) Focke is a kind of perennial herb which possesses many bioactivities such as antioxidative, anti-inflammatory properties and the ability to inhibit central nervous system. The whole plant of D. indica has been used as a folk medicine in treating fever, eczema, herpes zoster and traumatic injuries, etc. Recently, the chemical composition of D. indica has aroused researcher’s concern due to its excellent antitumor activity. Various active constituents, mainly terpenes, phenolic acids and flavonoids have been isolated from D. indica in recent years. Compared to the studies on soluble constitutions, the volatile components of D. indica have received little attention. Only several volatile components have been reported from an analysis on a steam distillation (SD) extraction of D. indica using gas chromatography–mass spectrometry. But the curative effect of D. indica may also be related to its volatile components. In this study, the volatile components of D. indica were studied by soxhlet extraction (SE), ultrasonic extraction (UAE), steam distillation and solid-phase microextraction (SPME). 66 compounds were identified totally, among which 47 compounds were first reported. Identification of these compounds should provide more information and expand the application of D. indica. Fritillaria is one of the largest genera in the plant family of Liliaceae. The bulbs of many Fritillaria species growing in China have been used as expectorant, anti-asthmatic and antitussive herbs with the Chinese name “Beimu” for more than 2000 years. Officially, herbal Beimu is the bulbs of 6 Fritillaria species documented in China Pharmacopoeia including Fritillariae cirrhosae bulbus (Chuan-Beimu, CB), Fritillariae thunbergii bulbus (Zhe-Beimu, ZB), Fritillariae ussuriensis bulbus (Ping-Beimu, PB), Fritillariae pallidiflorae bulbus (Yi-Beimu, YB) and Fritillariae hupehensis bulbus (Hubei-Beimu, HB). Among them, CB possesses characteristics of better therapy effects and lower production than others. Consequently, the value of CB is much higher than the other species, resulting in abundant adulterations with other species of the Fritillaria genus on the commercial markets. It is obviously that different species of fritillaria are cultivated from different geographical origins, and we speculate that the growing environment may also cause a significant difference in the volatile components of fritillaria. In chapter 3, the volatile components of four kinds of fritillaria were analyzed by solid-phase microextraction and gas chromatography-mass spectroscopy (SPME/GC-MS). The conditions for solid-phase microextraction were optimized by an orthogonal experimental design. Principal component analysis and hierarchical cluster analysis were performed to evaluate the GC-MS profiling of the different fritillaria species. The experimental results demonstrated that the same kinds of fritillaria has more similarity in the volatile components and SPME/ GC-MS can be considered as a distinguish method of different species of the fritillaria. Ovarian tumors can be divided into two categories, malignant tumor and the benign tumor. Mature teratoma of ovary is a typical benign tumor, which can be treated with surgery and achieve a quite high cure rate. Ovarian cancer is a kind of malignant tumor. Worldwide, about 125000 women succ |
| 语种 | 中文 |
| 公开日期 | 2016-05-03 |
| 源URL | [http://ir.ciac.jl.cn/handle/322003/64479]  |
| 专题 | 长春应用化学研究所_长春应用化学研究所知识产出_学位论文 |
| 推荐引用方式 GB/T 7714 | 王晨旭. 固相微萃取与气相色谱-质谱联用分析蛇莓、贝母和卵巢癌的挥发性组分[D]. 中国科学院长春应用化学研究所. 中国科学院研究生院. 2015. |
入库方式： OAI收割
来源：长春应用化学研究所
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