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研究生:邱佩玉
研究生(外文):chiu pei yu
論文名稱:製劑分析方法開發與柑橘傲才基原鑑定研究
指導教授:許順吉許順吉引用關係
指導教授(外文):Sheu Shuenn-Jyi
學位類別:碩士
校院名稱:國立臺灣師範大學
系所名稱:化學研究所
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:188
中文關鍵詞:柑橘
外文關鍵詞:citri
相關次數:
  • 被引用被引用:4
  • 點閱點閱:183
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  • 收藏至我的研究室書目清單書目收藏:0
高效液相層析(HPLC)和毛細管電泳(CE)同屬液相分離技術,都有多種分離模式,為目前最常用來定量中藥材指標成分的分析方法。本研究以這兩種儀器開發加味三黃錠製劑的分析方法,並比較其在複雜中藥製劑分析上的適宜性;陳皮及相關藥材,以中極性成分居多,本研究利用HPLC做為基原辨識的工具。
加味三黃錠由大黃、黃芩、黃連及白花蓮四種藥材所組成,可用CZE及MEKC方式分析。在CZE部分,利用硼酸鹽(pH=10.08)和氰甲烷調配的緩衝溶液,在40分鐘內分離其中九種純品成分(sennoside A, palmatine,berberine, rhein,aloe-emodin,emodin,chrysophanol,jatrorrhizine ,coptisine);在MEKC部分,利用含陰離子界面活性劑SC之硼酸鹽及磷酸鹽溶液和氰甲烷之緩衝溶液,在45分鐘內分析其中十九種純品成分(sennoside B,sennoside A,berberastine,baicalin,columbamine,jatrorrhizine,epiberberine,coptisine,oroxylin A-7-O-glucuronide,wogonin-7-O-glucronide,palmatine,berberine,baicalein,rhein,aloe-emodin,wogonin,oroxylin A,emodin,chrysophanol ),該分析條件之選擇和評估亦一併探討;應用於實際樣品分析,CE共可定量十九成分。在HPLC分析中,梯度沖提系統之流動相(A)為20mM KH2PO4 酸性水溶液(以10%H3PO4調整pH至2.47),流動相(B)為CH3CN/H2O = 80/20(V/V),在分離管柱Cosmosile 5C18-MS、偵測波長254及280 nm的條件下,於90分鐘內分析加味三黃錠的二十個純品成分(比CE方法多reserpine,但應用於製劑時,其量無法偵測);應用於實際樣品,共可分析十九成分。
檢討加味三黃錠之HPLC和CE的分析結果,發現在系統適宜性(包含再現性、回收率和偵測極限)、分析時間、基線穩定性及各吸收峰之理論板數等各有其優劣點,兩者應可相輔相成、搭配使用。
另外,本研究收集市售陳皮、青皮、橘紅、橘皮、橘白、橘紅皮及佛手等樣品共計81件,其中陳皮可分為廣陳皮、焦黑色陳皮及深褐色陳皮,青皮計有四化青皮及個青皮,橘紅與橘皮頗為相似,橘白與橘紅皮則是不同藥用部位,佛手為整個果實縱切。各樣品用HPLC分析,以umbelliferone ,citropten,narirutin,hesperidin,sinensetin,nobiletin,acacetin,tangeretin,5-demethyl-nobiletin及synephrine 等十個成分為指標,發現這些成分之總含量平均值依次為四化青皮、焦黑色陳皮、個青皮、橘紅、廣陳皮、橘紅皮、橘皮、橘白、深褐色陳皮及佛手,佛手含量極低不及其他藥材十分之一。以個別成分而言,synephrine在青皮含量較多,其中又以四化青皮為佳,在陳皮則完全沒有;umbelliferone則以焦黑色陳皮為多;naringin 及hesperidin以青皮及橘白含量比較多; sinensetin 及tangeretin 以橘紅皮為多;nobiletin 以焦黑色陳皮為多;acacetin 以深褐色陳皮為多;5-demethyl-nobiletin 以四化青皮為多;citropten則只存在佛手中。各不同藥材之LC指紋圖譜有明顯不同,但同類藥材則有明確規則性,可做為判別不同種類原料藥材的依據;原則上各藥材可用其中3至4個波峰積分面積比值分辨之。
Both high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) belong to the chromatographic technique and each has a plurality of separation modes. They are currently the most popularly used analysis methods for assaying the marker substances in Chinese herb drugs. In this study too methods for analyzing the Chinese herb formula Expanded Coptis and Rhubarb Combination Tablets (ECRCT) were developed, and their appropriateness in the analyses of complicated Chinese herbal formulas. Were compared Citri Leiocarpae Exocarpium and related herbs contain predominantly neutral constituents, wherefore this study used HPLC as a tool to identify the sources of the herbs.
ECRCT is composed of four herbs: Rhei Rhizoma, Scutellariae Radix, Coptidis Rhizoma and Rauwolfiae Verticillatae Radix, which can be analyzed by CZE and MEKC. The CZE method uses a buffer consisting of a borate (pH = 10.08) and acetonitrile, and can accomplish within 45 min. to analyze 9 pure constituents (sennoside A, palmatine, berberine, rhein, aloe-emodin, emodin, chrysophanol, jatrorrhizine and coptisine). The MEKC method uses a buffer comprising surfactant SC, a borate, a phosphate, and acetonitrile. The method can accomplish within 45 min. to assay 19 pure constituents (sennoside B, sennoside A, berberastine, baicalin, columbamine, jatrorrhizine, epiberberine, coptisine, oroxylin A-7-O-glucuronide, wogonin-7-O-glucuronide, palmatine, berberine, baicalein, rhein, aloe-emodin, wogonin, oroxyline A, emodin and chrysophanol). Selection and assessment of analysis conditions are also investigated and discussed When applied to analyzing read samples , CE can assay 19 constituents. A gradient eluent system in the HPLC method contains mobile phase (A), an acidic solution of 20 mM KH2PO4, and mobile phase (B), which is CH3CN/H2O = 80/20. The chromatographic column is Cosmosile 5C18-MS and the detection wavelengths are 254 and 280 nm. With these conditions, HPLC method can analyze 20 pure constituents in ECRCT within 90 min. (one more constituent, reserpine, than those the CE method can handle. However, in herbal preparations this extra constituent is too minute to be detected.) When applied to analysis of practical samples of herb preparations, the method can analyze 19 constituents.
In investigating the results of analyses of ECRCT by HPLC and CE, we have found that in the aspects of system suitability (including reproducibility, recovery and detection limit), analysis time, baseline stability and the theoretical plate numbers of the various peaks, both methods have superiorities and inferiorities, and that the two methods can support and accommodate each other if used jointly.
Besides, this study collected 81 samples of stale orange peel (Citri Leocarpae Exocarpium Vertustas), green orange peel (Citri Pericarpium Immaturus), red orange peel (Citri Exocarpium Rubrum), orange peel (Citri Leocarpae Exocarpium), white vascular tissue of orange fruit (Aurantii Vascular), ripe orange red peel (Citri Exocarpium Maturus Rubrum) and Buddha-hand orange (Citri Sarcodactyli Fructus). Among these articles, the stale orange peel is further divided into Cantonese stale orange peel, charred black stale orange peel and dark brown stale orange peel; the green orange peel includes szuhua green orange peel and go green orange peel; red orange peel and orange peel are quite similar; white vascular tissue of orange fruit and ripe orange red peel are derived from different parts of the orange fruit; and Buddha-hand orange is cut longitudinally. Each sample is analyzed by HPLC, using the ten constituents umbelliferone, citropten, narirutin, hesperidin, sinensetin, nobiletin, acacetin, tangeretin, 5-demethyl-nobiletin and synephrine as the marker substances. It is found that the average value of the total contents of these substances in the various articles is a decreasing in the order of szuhua green orange peel, charred black stale orange peel, go green orange peel, red orange peel, Cantonese stale orange peel, ripe orange red peel, orange peel, white vascular tissue of orange fruit, dark brown stale orange peel and Buddha-hand orange. Buddha-hand orange has the lowest value, about one tenth of those in the other articles. In terms of the individual constituents, synephrine exists in a comparatively higher amount in green orange peel, whereof the szuhua variety has the highest content and the stale orange peel does not contain it; umbelliferone is most abundant in charred black stale range peel; naringin and hesperidin are found to exist in higher quantities in green orange peel and white vascular tissue of orange fruit; sinensetin and tangeretin are higher in ripe orange red peel; nobiletin occupies the most in charred black stale orange peel; acacetin is higher in dark brown stale orange peel; 5-memethyl-nobiletin is present in higher quantity in szuhua green peel; while citropten exists only in Buddha-hand orange. The various articles of the herb materials show distinctly different LC fingerprints. And yet drug materials of the same category show a clear regularity in their fingerprint patterns, which can be used for identifying drug materials of different categories. As a rule, the peak-area ratios of 3 and 4 of the fingerprints can be used for the identification.
目 錄
圖目錄…………………………………………………………………….Ⅲ
表目錄……………………………………………………………………. Ⅵ
中文摘要…………………………………………………………………Ⅸ
英文摘要……………………………………………………………… .ⅩⅠ
第一章 緒論
第一節 前言…………………………………………………………..1
第二節 高效液相層析法……………………………………………..3
第三節 毛細管電泳分析法…………………………………………..5
第四節 分析條件參數與適宜性之評估……………………..………18
第二章 加味三黃錠之毛細管電泳分析
第一節 前言…………………………………………………………..30
第二節 實驗部分………………………………………………………34
第三節 結果與討論………………………..…………………………41
第三章 加味三黃錠之高效能液相層析法
第一節 前言…………………………………………………………..78
第二節 實驗部分………………………………………………………80
第三節 結果與討論………………………..………………………….86
第四章 市售柑橘藥材基原之化學辨識
第一節 前言…………………………………………………………..104
第二節 實驗部分……………………………………………………… 112
第三節 結果與討論………………………..………….…………….118
第五章 結論
第一節 HPLC與CE之比較…………………………..……………….158
第二節 柑橘藥材基原辨識…………………………………………..165
參考文獻…………………………………………………………………….171
附 錄一 加味三黃錠指標成分之UV吸收光譜圖……………………….180
附 錄二 柑橘類指標成分之UV吸收光譜圖……………………………..185
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