梔子.荊芥之成分分析及基原辨識
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2005
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高效液相層析儀（HPLC）為測定中藥指標成分最常用的分析工具，加上質譜儀（LC-MS）可達到定性與定量的雙重目標。本研究使用光二極陣列（PDA）及質譜儀為偵測器，開發梔子與荊芥藥材的分析適宜條件，測定各樣品中主要成分含量，以供基原辨識及品管的重要依據。
本研究分為三個部分，第一部分為分別開發梔子藥材之HPLC與LC-MS分析方法。梔子藥材為一清熱涼血的常用中藥，主要由環烯醚類、cyclohexenoids、酸類、黃酮類組成，其中又以環烯醚類為最主要的藥理活性成分。實驗結果顯示，以5C18-MSⅡ作為分離管柱，分別在磷酸系統與醋酸系統下，和氰甲烷、甲醇、水最佳比例作為有機沖提液，於240、440nm偵測下，均可成功的分析出十七個成份，依序為十個環烯醚類：geniposide(11)、gardenoside(7)、shanziside methylester(9)、deacetyl asperulosidic acid(1)、geniposidic acid(4)、genipin gentiobioside(10)、6”-p-coumaroyl genipin gentiobioside(15)、gardoside(2)、butoxygeniposide(3)、hydroxygeniposide(6)及三個酸類3,4-di-O-caffeoyl quinic acid(16)、chlorogenic acid(5)、crocin及三個cyclohexenoids類Jasminoide A(13)、Jasminoide B(8)、Jasminoide E(12) 及一個黃酮類rutin(17)。本方法之R.S.D良好，intraday 的滯留時間為0.38%至1.22%，面積比為0.04%至0.18﹪；interday 的滯留時間為0.47%至1.48﹪，面積比為0.11%至0.39﹪。
第二部分為運用開發的分析方法於梔子藥材之基原辨識、殼仁、成熟度上的含量研究，並將高效能液相層析圖譜中所獲得之資訊作雷達指紋圖。本研究中自順天堂與市售上共收集37批藥材，分別為山梔子30批，水梔子7批，品種上主要以genipin gentiobioside與shanziside比值作為標的，genipin gentiobioside/shanziside>2.0是山梔，genipin gentiobioside/
shanziside<2.0是水梔。此外，加以運用感應耦合電漿質譜儀(ICP-MS)進行66個微量元素分析，以山梔子Ca、Rb微量元素含量顯著高於水梔子。最後輔以多變量解析予以圖形化區辨兩品種，其中所使用之解析包括群聚分析、主成分分析以及區別分析，希望能於人工判別圖形及成分比較外，找到一個更有效率且有系統之鑑別方式，統計數據結果上顯示品種方面，可以8個有機成分和8個微量元素將所獲得的藥材加以分為兩類。
第三部分在開發荊芥藥材HPLC與LC-MS之分析方法。由於目前種植上藥材品種僅為一種，因此不同部位的使用療效與差異更顯出其重要性。實驗結果顯示，以5C18-MSⅡ作為分離管柱，分別在磷酸系統與醋酸系統下，和氰甲烷、甲醇、水最佳比例作為有機沖提液，於280nm偵測下，均可成功的分析出十個成份，依序為六個diosmetin(8)、hesperidin(5)、luteolin(9)、apigenin(10)、luteolin-7-O-glucoside(4)、apigenin-7-O-glucoside(6)及兩個酸類rosmarinic acid(3)、caffeic acid(1)及兩個揮發油類benzaldehyde(7)、cyclohexanone(2)。其R.S.D，intraday 的滯留時間為0.68%至1.31%，面積比為0.06%至0.11﹪；interday 的滯留時間為0.82%至1.46﹪，面積比為0.06%至0.17﹪。本研究自順天堂與市售共得9批荊芥穗、3批荊芥莖及13批荊芥全草，可發現其於含量上之高低均以荊芥全草>荊芥穗>荊芥莖，因此可推論出在市售方面應以全草及穗的使用上療效為較佳，此外尚加上對此一藥材的莖穗粗細及顏色對成熟度觀察作一變化圖。使用ICP-MS分析61個微量元素含量，發現以Ca、In、Eu、Tb、Ho、Er、Tm、Yb、Lu、Hf含量上之高低依然為荊芥全草>荊芥穗>荊芥莖順序排列，Ba含量則以荊芥全草>荊芥穗≒荊芥莖，因此可獲知不論在有機成分或微量元素含量分析對於藥材之整體運用上，以使用穗及全草為較佳。最後輔以多變量解析，可獲知以2個有機成分與8個微量元素將所獲的藥材清楚將荊芥穗與荊芥莖作一良好的區別，僅於全草上則涵蓋此兩分類中。
High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometer (LC-MS) are the most common tools for analyzing the marker substances in Chinese herbs. In this study, photodiode array (PDA) and mass spectrometer were used as detectors and appropriate analyzing methods had been developed to determine the major components and their amount contained in each sample of Gardenia jasminoides Ellis. and Schizonepeta tenuifolia Briq.. The results provide an important basis for chemotaxonomic identification as well as quality control. This study consists of three parts: first, were established the analyzing methods of HPLC and LC-MS for iridoid glycosides, cyclohexenoids, acids, and flavones in Gardenia jasminoides Ellis.. With 5C18-MSⅡ column, phosphate and acetate buffers,the contents of seventeen components are measured under 240 and 440 nm: ten iridoid glycosides including geniposide (11), gardenoside (7), shanziside methylester (9), deacetyl asperulosidic acid (1), geniposidic acid (4), genipin gentiobioside (10), 6”-p-coumaroyl genipin gentiobioside (15), gardoside (2), butoxygeniposide (3), and hydroxygeniposide (6); three acids including 4-di-O-caffeoyl quinic acid (16), chlorogenic acid (5), and crocin; three cyclohexenoids including Jasminoide A (13), Jasminoide B (8), and Jasminoide E (12); one flavones, rutin (17).The method shows good reproducibility for retention times and peak-area ratios in either marker substances or plant extracts. Second, the analyse described above were used for the chemotaxonomic identification and the comparison quantitative of shell and maturity for Gardenia jasminoides Ellis.. In this study, 37 samples were collected from Sun Ten Pharmaceutical Co. Ltd. and the herbal shops in Taiwan, including 30 samples of Gardenia jasminoides Ellis. and 7 samples of Gardenia jasminoides Ellis. var. grandiflora Nakai.. It is shown that the ratio of genipin gentiobioside/shanziside > 2.0 belongs to Gardenia jasminoides Ellis. and genipin gentiobioside/shanziside< 2.0 is Gardenia jasminoides Ellis. var. grandiflora Nakai. In addition, inductively coupled plasma-mass spectrometer (ICP-MS) is used to analyze 66 trace elements. The amount of Ca and Rb in Gardenia jasminoides Ellis. is significantly higher than that in Gardenia jasminoides Ellis. var. grandiflora Nakai. A multivariate analysis including cluster, major component, and differentiation analysis was also used. In stead of comparing with their components manually, this study expects to look for amore efficient method in distinguishing Gardenia jasminoides Ellis. and var. grandiflora Nakai..The resulting statistics shows that those samples can be categorized into two groups by 8 organic components and 8 trace elements. Finally, HPLC and LC-MS were also developed for analyzing Schizonepeta tenuifolia Briq. samples. With 5C18-MSⅡ column, phosphate and acetate buffers, the contents of ten components are measured under 280 nm: they are diosmetin (8), hesperidin (5), luteolin (9), apigenin (10), luteolin-7-O-glucoside (4), apigenin-7-O-glucoside (6), rosmarinic acid (3), caffeic acid (1), benzaldehyde (7), and cyclohexanone (2). The method shows good reproducibility for retention times and peak-area ratios in either marker substances or plant extracts.In this study, 25 samples were collected from Sun Ten Pharmaceutical Co. Ltd. and the herbal shops in Taiwan ,including 9 batches of Schizonepeta tenuifolia Briq. tassels, 3 batches of stems, and 13 batches of whole trunks to determine the amount contained. From the most to the least is: whole trunks＞tassels＞stems. We therefore conclude that the whole trunks and tassels of Schizonepeta tenuifolia Briq. are more effective in remedy. In ICP-MS analysis, among 61 trace elements, the amount of Ca, In, Eu, Tb, Ho, Er, Tm, Yb, Lu, and Hf from more to less is: whole trunks＞tassels＞stems. The quantitative analysis of Ba is: whole trunks>tassels≒stems. It is thus found that tassels and whole trunks are more effective in remedy after analysis of organic components and trace elements. A multivariate analysis was also used. In stead of comparing with their components manually, this study expects to look for a more efficient method in distinguishing Schizonepeta tenuifolia Briq. tassels from stems. The resulting statistics shows that those samples can be categorized into two groups by 2 organic components and 8 trace elements.
High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometer (LC-MS) are the most common tools for analyzing the marker substances in Chinese herbs. In this study, photodiode array (PDA) and mass spectrometer were used as detectors and appropriate analyzing methods had been developed to determine the major components and their amount contained in each sample of Gardenia jasminoides Ellis. and Schizonepeta tenuifolia Briq.. The results provide an important basis for chemotaxonomic identification as well as quality control. This study consists of three parts: first, were established the analyzing methods of HPLC and LC-MS for iridoid glycosides, cyclohexenoids, acids, and flavones in Gardenia jasminoides Ellis.. With 5C18-MSⅡ column, phosphate and acetate buffers,the contents of seventeen components are measured under 240 and 440 nm: ten iridoid glycosides including geniposide (11), gardenoside (7), shanziside methylester (9), deacetyl asperulosidic acid (1), geniposidic acid (4), genipin gentiobioside (10), 6”-p-coumaroyl genipin gentiobioside (15), gardoside (2), butoxygeniposide (3), and hydroxygeniposide (6); three acids including 4-di-O-caffeoyl quinic acid (16), chlorogenic acid (5), and crocin; three cyclohexenoids including Jasminoide A (13), Jasminoide B (8), and Jasminoide E (12); one flavones, rutin (17).The method shows good reproducibility for retention times and peak-area ratios in either marker substances or plant extracts. Second, the analyse described above were used for the chemotaxonomic identification and the comparison quantitative of shell and maturity for Gardenia jasminoides Ellis.. In this study, 37 samples were collected from Sun Ten Pharmaceutical Co. Ltd. and the herbal shops in Taiwan, including 30 samples of Gardenia jasminoides Ellis. and 7 samples of Gardenia jasminoides Ellis. var. grandiflora Nakai.. It is shown that the ratio of genipin gentiobioside/shanziside > 2.0 belongs to Gardenia jasminoides Ellis. and genipin gentiobioside/shanziside< 2.0 is Gardenia jasminoides Ellis. var. grandiflora Nakai. In addition, inductively coupled plasma-mass spectrometer (ICP-MS) is used to analyze 66 trace elements. The amount of Ca and Rb in Gardenia jasminoides Ellis. is significantly higher than that in Gardenia jasminoides Ellis. var. grandiflora Nakai. A multivariate analysis including cluster, major component, and differentiation analysis was also used. In stead of comparing with their components manually, this study expects to look for amore efficient method in distinguishing Gardenia jasminoides Ellis. and var. grandiflora Nakai..The resulting statistics shows that those samples can be categorized into two groups by 8 organic components and 8 trace elements. Finally, HPLC and LC-MS were also developed for analyzing Schizonepeta tenuifolia Briq. samples. With 5C18-MSⅡ column, phosphate and acetate buffers, the contents of ten components are measured under 280 nm: they are diosmetin (8), hesperidin (5), luteolin (9), apigenin (10), luteolin-7-O-glucoside (4), apigenin-7-O-glucoside (6), rosmarinic acid (3), caffeic acid (1), benzaldehyde (7), and cyclohexanone (2). The method shows good reproducibility for retention times and peak-area ratios in either marker substances or plant extracts.In this study, 25 samples were collected from Sun Ten Pharmaceutical Co. Ltd. and the herbal shops in Taiwan ,including 9 batches of Schizonepeta tenuifolia Briq. tassels, 3 batches of stems, and 13 batches of whole trunks to determine the amount contained. From the most to the least is: whole trunks＞tassels＞stems. We therefore conclude that the whole trunks and tassels of Schizonepeta tenuifolia Briq. are more effective in remedy. In ICP-MS analysis, among 61 trace elements, the amount of Ca, In, Eu, Tb, Ho, Er, Tm, Yb, Lu, and Hf from more to less is: whole trunks＞tassels＞stems. The quantitative analysis of Ba is: whole trunks>tassels≒stems. It is thus found that tassels and whole trunks are more effective in remedy after analysis of organic components and trace elements. A multivariate analysis was also used. In stead of comparing with their components manually, this study expects to look for a more efficient method in distinguishing Schizonepeta tenuifolia Briq. tassels from stems. The resulting statistics shows that those samples can be categorized into two groups by 2 organic components and 8 trace elements.
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梔子, 荊芥