白屈菜质量评价研究
摘要
本文以常用中药白屈菜为研究对象,采用薄层色谱、液相色谱、特征图谱等方法对收集于中国不同产地的20批白屈菜药材进行了质量标准研究。建立了药材中白屈菜红碱、白屈菜碱、血根碱、原阿片碱、盐酸小檗碱、黄连碱6个活性成分的薄层色谱鉴别方法。用紫外-可见分光光度法测定了药材中总生物碱的含量,并建立了原阿片碱、白屈菜碱、黄连碱、四氢黄连碱、血根碱、盐酸小檗碱、白屈菜红碱等7个生物碱成分的UPLC含量测定方法。此外还建立了20批药材UPLC特征图谱,用Q-TOF-MS鉴别了17个共有峰中的14个生物碱类成分,其中6个由对照品对照确认,8个由质谱数据推测而得,并在此基础上利用MS-MS信息初步推测了白屈菜中三大类生物碱成分的裂解方式。从而建立了白屈菜药材的质量标准。
本论文共分两章。
第一章综述综述了白屈菜的质量研究概况,主要内容有:本草考证、化学成分、药理作用、质量控制研究现状。
第二章白屈菜药材质量评价研究共分七节,简述如下:
第一节白屈菜样品采集简述了本文所用的白屈菜药材的收集情况。
第二节白屈菜显微鉴别研究对白屈菜全草进行详细的显微鉴别,为白屈菜真伪鉴别提供了一定客观依据,建立了白屈菜药材质量标准【鉴别】(1)项。
第三节白屈菜薄层色谱鉴别研究进行了一系列方法学考察,包括样品处理方法、展开剂的选择与优化、显色方法、点样量等,最终确定了样品中以对照药材和白屈菜红碱、白屈菜碱、血根碱、原阿片碱、盐酸小檗碱、黄连碱为对照的薄层色谱鉴别方法,建立了白屈菜药材质量标准【鉴别】(2)项。
第四节白屈菜水分、灰分、浸出物检查考察了不同检查方法,最终选择烘干法(第一法)测定白屈菜水分,以稀乙醇为溶剂测定白屈菜浸出物。制定其水分不得过13.0%,总灰分不得过12.0%,酸不溶性灰分不得过0.6%,浸出物不得低于17.0%。
第五节白屈菜生物碱类成分的含量测定研究
此节包括两部分内容。
内容一紫外-可见分光光度法测定白屈菜药材中总生物碱含量
采用紫外-可见分光光度法对白屈菜药材中总生物碱的含量进行了测定,并考察了该方法的可行性。该方法简便,干扰少,结果可靠。20批药材中总生物碱最高含量为32.67mg/g,最低含量为23.25 mg/g,平均含量为26.02 mg/g。
内容二UPLC同时测定白屈菜中7个主要生物碱成分的含量
色谱柱:ACQUITY UPLC BEH C_(18)(2.1×100 mm,1.7μm)。流动相:乙腈(A)-10 mmol/L醋酸铵缓冲液(用醋酸调节至pH 3.0)(B);梯度洗脱;柱温35℃;流速0.4ml/min;进样量:1μl。检测波长:240 nm用于原阿片碱、白屈菜碱、四氢黄连碱和白屈菜红碱,270nm用于黄连碱、血根碱和盐酸小檗碱。该条件下线性关系r≥0.9992,高、中、低三个水平回收率在95.55%至103.08%之间。用该方法测定7个生物碱的含量,快速、简便、准确、重复性好、溶剂消耗少且分离度高,为白屈菜定量分析提供了一定依据,有利于白屈菜质量控制研究。
第六节白屈菜UPLC特征图谱建立及生物碱类成分QTOF-MS分析研究采用UPLC建立了白屈菜药材的生物碱类成分特征图谱,分析时间在30分钟以内。20批白屈菜样本之间相似度大于0.90。获得17个共有峰。通过QTOF-MS技术对其共有峰进行分析,鉴定出14个生物碱成分,其中6个通过对照品对照确认,8个通过质谱推测,初步探讨了生物碱的裂解规律。
第七节白屈菜质量标准草案根据本文研究,拟定起草了一份白屈菜药材的全面质量标准草案。
This paper aimed at the 20 batches of Chelidonium majus L. which were collected from different location in China. A new method of TLC was established to identify the six alkaloids (protopine, chelidonine, coptisine, stylopine, sanguinarine, berberine and chelerythrine). An UPLC method was developed to determine protopine, chelidonine, coptisine, stylopine, sanguinarine, berberine and chelerythrine and the total alkaloids were also' determined by vis-spectrophotometry. Besides the UPLC specific chromatogram of Chelidonium majus L. was established and analyze the alkaloids compositions by Q-TOF-MS. There were 17 common peaks. Fourteen alkaloids compositions were identified. Among them, 6 compositions were identified by direct comparison with the reference substances and others were identified by MS and MS2 data. These methods were useful for the evaluation of the quality of Chelidonium majus L. systematically and roundly.
This dissertation is divided into two major parts.
The first part was a detailed review about the latest research of quality evaluation for Chelidonium majus L., including herbal textual research, chemical constituents, pharmacological effects and quality standard.
In the second part, there were seven chapters. Studied on the quality evaluation of Chelidonium majus L.
In the first chapter, it was the informations of the investigated samples.
In the second chapter, it was studied on the microscopic identification of Chelidonium majus L. and builded the [Identification](1) item.
In the third chapter, it was studied on TLC. In order to obtain TLC chromatograms with better resolution for six alkaloids, different extracting solvents, chromatographic conditions and colorimetric methods were compared. Then, the TLC method used for Chelidonium majus L. identification has been established named [Identification](2).
In the fourth chapter, it was studied on the content determination of water, ash content and extractives. Different methods were optimized. Then the method 1 (Drying in oven method) was used for determination of water in crude drugs containing no or scarcely any volatile constituents. The limitation of water was not exceed 13.0%, 12.0% for total ash, 0.6% for acid-insoluble ash and the limitation of extractives was not less than 17.0%.
In the fifth chapter, it was studied on the content determination of alkaloids.
This chapter included two parts.
Part one was studied on the content determination of total alkaloids. It was performed on vis-spectrophotometry and the method was simple and showed good precision, repeatability and accuracy. The total alkaloids contents was23.25 mg/g～32.67mg/g and the average were 26.02mg/g.
Part two was studied on the content determination of seven main active alkaloids. The UPLC method was performed on an ACQUITY UPLC BEH C18 (2.1×100 mm, 1.7μm) column. The mobile phase was composed of (A) acetonitrile and (B) 10 mmol/L mmonium acetate (adjusted to pH 3.0 with acetic acid) with a linear gradient elution. The flow rate of the mobile phase was 0.4 ml/min and the column temperature was maintained at 35℃. The injection volume was 1μl. Detection wavelength was set at 240 nm for protopine, chelidonine, stylopine, chelerythrine and 270nm for coptisine, sanguinarine, berberine. Good linear behaviors(r > 0.9992) over the investigated concentration ranges were observed for all the analytes. The recoveries (low, medium and high levels) were 95.55%～103.08%. The UPLC-PDA method was developed for simultaneously determination of 7 alkaloids in Chelidonium majus L., it showed good precision, repeatability, accuracy, fast analysis, acceptable resolution and less solvent consumption. The method was useful for authenticity controlling the quality of Chelidonium majus L.
In the sixth chapter, the method of fingerprint of alkaloids in Chelidonium majus L. was setting up by UPLC-PDA. The analysis time was 30 min. The results of similarity evaluation showed that the similarity was more than 0.90. There were 17 common peaks. Besides the analysis of alkaloids compositions was performed by Q-TOF-MS. Fourteen alkaloids compositions were identified. Among them, 6 compositions were identified by direct comparison with the reference substances and others were identified by MS and MS2 data. And the fragmentation patterns of different alkaloids were inferred.
In the seventh chapter, it was the protocol of the quality standard of Chelidonium majus L. which we builded based on the paper.
本论文共分两章。
第一章综述综述了白屈菜的质量研究概况,主要内容有:本草考证、化学成分、药理作用、质量控制研究现状。
第二章白屈菜药材质量评价研究共分七节,简述如下:
第一节白屈菜样品采集简述了本文所用的白屈菜药材的收集情况。
第二节白屈菜显微鉴别研究对白屈菜全草进行详细的显微鉴别,为白屈菜真伪鉴别提供了一定客观依据,建立了白屈菜药材质量标准【鉴别】(1)项。
第三节白屈菜薄层色谱鉴别研究进行了一系列方法学考察,包括样品处理方法、展开剂的选择与优化、显色方法、点样量等,最终确定了样品中以对照药材和白屈菜红碱、白屈菜碱、血根碱、原阿片碱、盐酸小檗碱、黄连碱为对照的薄层色谱鉴别方法,建立了白屈菜药材质量标准【鉴别】(2)项。
第四节白屈菜水分、灰分、浸出物检查考察了不同检查方法,最终选择烘干法(第一法)测定白屈菜水分,以稀乙醇为溶剂测定白屈菜浸出物。制定其水分不得过13.0%,总灰分不得过12.0%,酸不溶性灰分不得过0.6%,浸出物不得低于17.0%。
第五节白屈菜生物碱类成分的含量测定研究
此节包括两部分内容。
内容一紫外-可见分光光度法测定白屈菜药材中总生物碱含量
采用紫外-可见分光光度法对白屈菜药材中总生物碱的含量进行了测定,并考察了该方法的可行性。该方法简便,干扰少,结果可靠。20批药材中总生物碱最高含量为32.67mg/g,最低含量为23.25 mg/g,平均含量为26.02 mg/g。
内容二UPLC同时测定白屈菜中7个主要生物碱成分的含量
色谱柱:ACQUITY UPLC BEH C_(18)(2.1×100 mm,1.7μm)。流动相:乙腈(A)-10 mmol/L醋酸铵缓冲液(用醋酸调节至pH 3.0)(B);梯度洗脱;柱温35℃;流速0.4ml/min;进样量:1μl。检测波长:240 nm用于原阿片碱、白屈菜碱、四氢黄连碱和白屈菜红碱,270nm用于黄连碱、血根碱和盐酸小檗碱。该条件下线性关系r≥0.9992,高、中、低三个水平回收率在95.55%至103.08%之间。用该方法测定7个生物碱的含量,快速、简便、准确、重复性好、溶剂消耗少且分离度高,为白屈菜定量分析提供了一定依据,有利于白屈菜质量控制研究。
第六节白屈菜UPLC特征图谱建立及生物碱类成分QTOF-MS分析研究采用UPLC建立了白屈菜药材的生物碱类成分特征图谱,分析时间在30分钟以内。20批白屈菜样本之间相似度大于0.90。获得17个共有峰。通过QTOF-MS技术对其共有峰进行分析,鉴定出14个生物碱成分,其中6个通过对照品对照确认,8个通过质谱推测,初步探讨了生物碱的裂解规律。
第七节白屈菜质量标准草案根据本文研究,拟定起草了一份白屈菜药材的全面质量标准草案。
This paper aimed at the 20 batches of Chelidonium majus L. which were collected from different location in China. A new method of TLC was established to identify the six alkaloids (protopine, chelidonine, coptisine, stylopine, sanguinarine, berberine and chelerythrine). An UPLC method was developed to determine protopine, chelidonine, coptisine, stylopine, sanguinarine, berberine and chelerythrine and the total alkaloids were also' determined by vis-spectrophotometry. Besides the UPLC specific chromatogram of Chelidonium majus L. was established and analyze the alkaloids compositions by Q-TOF-MS. There were 17 common peaks. Fourteen alkaloids compositions were identified. Among them, 6 compositions were identified by direct comparison with the reference substances and others were identified by MS and MS2 data. These methods were useful for the evaluation of the quality of Chelidonium majus L. systematically and roundly.
This dissertation is divided into two major parts.
The first part was a detailed review about the latest research of quality evaluation for Chelidonium majus L., including herbal textual research, chemical constituents, pharmacological effects and quality standard.
In the second part, there were seven chapters. Studied on the quality evaluation of Chelidonium majus L.
In the first chapter, it was the informations of the investigated samples.
In the second chapter, it was studied on the microscopic identification of Chelidonium majus L. and builded the [Identification](1) item.
In the third chapter, it was studied on TLC. In order to obtain TLC chromatograms with better resolution for six alkaloids, different extracting solvents, chromatographic conditions and colorimetric methods were compared. Then, the TLC method used for Chelidonium majus L. identification has been established named [Identification](2).
In the fourth chapter, it was studied on the content determination of water, ash content and extractives. Different methods were optimized. Then the method 1 (Drying in oven method) was used for determination of water in crude drugs containing no or scarcely any volatile constituents. The limitation of water was not exceed 13.0%, 12.0% for total ash, 0.6% for acid-insoluble ash and the limitation of extractives was not less than 17.0%.
In the fifth chapter, it was studied on the content determination of alkaloids.
This chapter included two parts.
Part one was studied on the content determination of total alkaloids. It was performed on vis-spectrophotometry and the method was simple and showed good precision, repeatability and accuracy. The total alkaloids contents was23.25 mg/g～32.67mg/g and the average were 26.02mg/g.
Part two was studied on the content determination of seven main active alkaloids. The UPLC method was performed on an ACQUITY UPLC BEH C18 (2.1×100 mm, 1.7μm) column. The mobile phase was composed of (A) acetonitrile and (B) 10 mmol/L mmonium acetate (adjusted to pH 3.0 with acetic acid) with a linear gradient elution. The flow rate of the mobile phase was 0.4 ml/min and the column temperature was maintained at 35℃. The injection volume was 1μl. Detection wavelength was set at 240 nm for protopine, chelidonine, stylopine, chelerythrine and 270nm for coptisine, sanguinarine, berberine. Good linear behaviors(r > 0.9992) over the investigated concentration ranges were observed for all the analytes. The recoveries (low, medium and high levels) were 95.55%～103.08%. The UPLC-PDA method was developed for simultaneously determination of 7 alkaloids in Chelidonium majus L., it showed good precision, repeatability, accuracy, fast analysis, acceptable resolution and less solvent consumption. The method was useful for authenticity controlling the quality of Chelidonium majus L.
In the sixth chapter, the method of fingerprint of alkaloids in Chelidonium majus L. was setting up by UPLC-PDA. The analysis time was 30 min. The results of similarity evaluation showed that the similarity was more than 0.90. There were 17 common peaks. Besides the analysis of alkaloids compositions was performed by Q-TOF-MS. Fourteen alkaloids compositions were identified. Among them, 6 compositions were identified by direct comparison with the reference substances and others were identified by MS and MS2 data. And the fragmentation patterns of different alkaloids were inferred.
In the seventh chapter, it was the protocol of the quality standard of Chelidonium majus L. which we builded based on the paper.
引文
[1]张焱.浅谈规范中药质量标准的迫切性[J].山西中医:34.
[2]赵维胜,杨占玉.浅谈中药质量标准的现代化[J].医学信息,2009,(11):2608-2611.
[3]胡世林.中国地道药材[M].哈尔滨:黑龙江科学技术出版社.1989:97.
[4]任德权.中药质量控制的里程碑--中药特征图谱[J].中成药.2001,23(1):1.
[5]谢培山.中药制剂色谱特征图谱鉴别[J].中成药.2002,22(6):193.
[6]肖树雄,郑剑红,杨文红.中药检验中特征图谱应用的新进展[J].GAP研究与实践.2002,2(1):41.
[7]中华本草委员会.中华本草[M],第9卷.上海科技出版社.1999:616.
[8]中国人民共和国药典委员会.中华人民共和国药典[S],第一部.化学工业出版社.1977:176.
[9]徐国钧,王强.中草药彩色图谱[M].第三版.福建科技出版社.2006:764.
[10]Taborska E,Bochorakova H,Dostal J,Paulova H.The greater celandine(Chelidonium majus L.)--review of present knowledge[J].Ceska Slov Farm.1995 Apr;44(2):71-75.
[11]黄松,杜方麓.罂粟科白屈菜族的化学成分及植物化学分类依据的研究进展[J].湖南中医药导报.2002,8(10):582-584.
[12]郭靖,吴连举,刘继永.白屈菜的药理及临床应用[J].特种经济植物.2001,(10):31.
[13]ManskeR H F.The Alkaloids Chemistry and Pharmaeology.Vol.15.New Yrok:Aeademie Press Ine,1975:207-243.
[14]何志敏,佟继铭,宫凤春.白屈菜碱镇痛作用研究[J].中草药.2003.9,(34):834-838.
[15]佟继铭,刘玉玲,陈光辉,等.白屈菜总生物碱镇咳平喘作用[J].承德医学院学报,2001,18(4):277.
[16]左金梁.白屈菜药材及其制剂的质量控制方法研究[D].沈阳:沈阳药科大学.2008.
[17]宗永立,刘艳平.白屈菜红碱对人胃癌BGC823细胞的增殖抑制和凋亡诱导作用[J].中草药.2006.7,37(7):1054-1056.
[18]Lanvers-Kaminsky C,Nolting DM,K(o|¨)ster J.et.al,Anticancer Drugs.17(2006)1025-1030.
[19]Vavreckova C,Gawlik I,M(u|¨)ller K,Benzophenanthridine alkaloids of Chelidonium majus;Ⅱ.Potent inhibitory action against the growth of human keratinocytes[J].Planta Med.1996 Dec;62(6):491-494.
[20]宋立人,洪询,丁绪亮,减载阳.现代中药学大辞典[M].北京:人民卫生出版社,2001:697.
[21]王本样.现代中药药理与临床[M].天津:天津科技翻译出版公司,2004:522.
[22]杨瑞林,魏学智,毕润成.山西霍山罂粟科植物花粉形态[J].广西植物,2003,23(3):231-232
[23]Walterova D,Preininger V,Sirnanek V.Qualitative and quantitative isotaehophoretic analysis of some quaternary isoquinoline alkaloids[J].Planata Med,1984,50(2):149.
[24]Dzido T,Jusiak L,Soezewinski E.Analysis of alkaloid extraet of Chelidonium majus by TLC and micro-bore HPLC[J].Chem Anal,1986,31(1):135.
[25]Bugatti,C.,Colombo,M.L.,Tome,F.High-performance liquid chromatographic separation of quaternary alkaloids of Chelidonium majus L.roots[J].J.Chromatogr.1987,393,312-316.
[26]Han,L.F.,Nowicky,W.,Gutmann,V.,Reversed-phase high-performance liquid chromatographic separation of tertiary and quatemary alkaloids from Chelidoaium majus L[J].J.Chromatogr.1991,543,123-128.
[27]牛长群,何丽一.白屈菜中异哇琳生物碱的高效薄层分离及荧光扫描法[J].药学学报,1991,27(1):69.
[28]Taborska,E.,Bochorakova,H.,Paulova,H.,Dostal,Separation of Alkaloids in Chelidonium majus by Reversed Phase HPLC[J].J.,Planta Med.1994,60,380-381.
[29]Chang-qun,Li-yi He,Determination of isoquinoline alkaloids in Chelidonium majus L.by ion-pair high-performance liquid chromatography[J].J Chromatogr.1991,542,193-199.
[30]Rey,J.P.,Levesque,J.,Pousset,J.L.,Analytical studies of dl-stylopine in Chelidonium majus L.using high-performance liquid chromatography[J].J.Chromatogr.1993,641,180-183.
[31]Kursinszki,L.,Sark(o|¨)zi,A,Kery,A,Sz(o|¨)ke.E,Improved RP-HPLC Method for Analysis of Isoquinoline Alkaloids in Extracts of Chelidonium majus[J].J.Chromatogr,2006,63,S131-S135.
[32]Sark(o|¨)zi,A.,Janicsak,G.,Kursinszki L.,Kery.A.Alkaloid Composition of Chelidonium majus L.Studied by Different Chromatographic Techniques[J].J.Chromatogr,2006,63,S81-S86.
[33]Jana Suchomelova,Hana Bochoiakova,Hana Paulova,Pavel Musil,Eva Tatborska,HPLC quantification of seven quaternary benzo[c]phenanthridine alkaloids in six species of the family Papaveraceae[J].J.Pharm.Biomed.Anal.44(2007) 283-287.
[34]左金梁,白璐,宋欣鑫,等.RP-HPLC法同时测定白屈菜药材中血根碱、小檗碱和白屈菜红碱的含量[J].药物分析杂志.2008,28(6):903-905
[35]曲彩红,陶玲,黄际薇,等.高效液相色谱法测定白屈菜注射液中白屈菜碱的含量[J].中国医院药学杂志.2005,25(7):623-625
[36]韩海.高效液相色谱法同时测定胃痛舒胶囊中原阿片碱和白屈菜碱含量[J].时珍国医国药.2005,16(11):1092-1093
[37]陶玲,曲彩红,黄际薇,等.白屈菜注射液HPLC特征图谱研究[J].中药材,2004.5,27(5):376-377
[38]李晓蒙,朱盛华,廖华卫.白屈菜药材HPLC特征图谱研究[J].天然产物研究与开发.2002.6(14):33
[39]Bo Shao,Hong-zhu Guo,Ya-jun Cui,Ai-hua Liu,Hai-lan Yu,Hui Guo,Man Xu,De-an Guo,Simultaneous determination of six major stilbenes and flavonoids in Smilax china by high performance liquid chromatography[J].J.Pharm.Biomed.Anal.44(2007) 737-742.
[40]Xiao-jie Tan,Qing Li,Xiao-hui Chen,Zhi-wei Wang,Zheng-yuan Shi,Kai-shun Bi,Ying Jia,Simultaneous determination of 13 bioactive compounds in Herba Artemisiae Scopariae(Yin Chen) from different harvest seasons by HPLC-DAD[J].J.Pharm.Biomed.Anal.47(2008) 847-853.
[41]Yang Zhao,Zhangwan Li,Xin Zhou,Zongwei Cai,Xiaojian Gong,Chanyuan Zhou.Quality evaluation of Evodia rutaecarpa(Juss.) Benth by high performance liquid chromatography with photodiode-array detection[J].J.Pharm.Biomed.Anal.48(2008)1230-1236.
[42]Mei Liu,Yongguo Li,Guixin Chou,Xuemei Cheng,Mian Zhang,Zhengtao Wang.Extraction and ultra-performance liquid chromatography of hydrophilic and lipophilic bioactive components in a Chinese herb Radix Salviae Miltiorrhizae[J].J.Chromatogr.A 1157(2007) 51-55.
[43]Li Yang,Aizhen Xiong,Yuqi He,Zaiyong Wang,Changhong Wang,Zhengtao Wang,et.al.Bile Acids Metabonomic Study on the CC_(14)- and a-Naphthylisothiocyanate-Induced Animal Models: Quantitative Analysis of 22 Bile Acids by Ultraperformance Liquid Chromatography-Mass Spectrometry[J]. Chem. Res. Toxicol. 21 (2008) 2280-2288.
[44]J. Guan, CM. Lai, S.P. Li, A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography[J]. J. Pharm. Biomed. Anal. 44 (2007) 996-1000.
[45]Szabolcs Fekete, Jeno Fekete, Katalin Ganzler. Validated UPLC method for the fast and sensitive determination of steroid residues in support of cleaning validation in formulation area[J]. J. Pharm. Biomed. Anal. 49 (2009) 833-838.
[1]傅泉炎.中药含水量与中药仓储质量的关系[J].传统医药.2002,11(5):70.
[2]王淑慧.加强中药材灰分检测[J].现代中药研究与实践.1997.11(3):44.
[3]徐晓光,王宏,董晓辉.中华人民共和国国家标准《人参加工产品分等质量标准》(三)对人参中浸出物含量标准及卫生学标准的研究与讨论[J].人参研究.1995.(2):33.
[4]严辉,段金廒,钱大玮,宿树兰,宋秉生,何子清.我国不同产地当归药材质量的分析与评价[J].中草药.2009.12,40(12):1988-1992.
[5]孙振球.医学统计学(第二版)[M].北京:人民卫生出版社.2006.
[6]Matile.Philipe.Loealization of alkaloids and mechanism of their accumulation in vacuoles of Cheidonium majus laticifers.Nova Acta Leopold Suppl,1976.7:139
[7]冯瑞芝,连文琰,傅桂香,等.罂粟科白屈菜族的化学分类及资源利用[J].植物分类学报.1985,23(1):36.
[8]Slavik J,Slavikova L.Minor alkaloids from Cheldonum majus L[J].Collection Czechoslov Chem Commun,1977,42:2686-2693.
[9]李亚伟,方起程.籽纹紫堇中的异喹啉类生物碱[J].中草药,1991,22(11):486.
[10]王恒山,杨海荣.迭裂黄堇生物碱的研究[J].天然产物研究与开发.1997,(1):37-39.
[11]杜方麓,陈胜璜,阳长明,等.血水草化学成分研究[J].中草药.1993,24(4):177.
[12]于敏,陈卫红,焦连庆.等.白屈菜的研究进展[J].特产研究.2008(2):76-78.
[13]李立军,再帕尔·阿不力孜,项赟,等.黄连总碱的MS/MS及LC-MS/MS研究[J].质谱学报.21(3,4):81-82.
[14]匡海学,董小萍,石任兵.中药化学(第一版).北京:中国中医药出版社.2003:341.
[2]赵维胜,杨占玉.浅谈中药质量标准的现代化[J].医学信息,2009,(11):2608-2611.
[3]胡世林.中国地道药材[M].哈尔滨:黑龙江科学技术出版社.1989:97.
[4]任德权.中药质量控制的里程碑--中药特征图谱[J].中成药.2001,23(1):1.
[5]谢培山.中药制剂色谱特征图谱鉴别[J].中成药.2002,22(6):193.
[6]肖树雄,郑剑红,杨文红.中药检验中特征图谱应用的新进展[J].GAP研究与实践.2002,2(1):41.
[7]中华本草委员会.中华本草[M],第9卷.上海科技出版社.1999:616.
[8]中国人民共和国药典委员会.中华人民共和国药典[S],第一部.化学工业出版社.1977:176.
[9]徐国钧,王强.中草药彩色图谱[M].第三版.福建科技出版社.2006:764.
[10]Taborska E,Bochorakova H,Dostal J,Paulova H.The greater celandine(Chelidonium majus L.)--review of present knowledge[J].Ceska Slov Farm.1995 Apr;44(2):71-75.
[11]黄松,杜方麓.罂粟科白屈菜族的化学成分及植物化学分类依据的研究进展[J].湖南中医药导报.2002,8(10):582-584.
[12]郭靖,吴连举,刘继永.白屈菜的药理及临床应用[J].特种经济植物.2001,(10):31.
[13]ManskeR H F.The Alkaloids Chemistry and Pharmaeology.Vol.15.New Yrok:Aeademie Press Ine,1975:207-243.
[14]何志敏,佟继铭,宫凤春.白屈菜碱镇痛作用研究[J].中草药.2003.9,(34):834-838.
[15]佟继铭,刘玉玲,陈光辉,等.白屈菜总生物碱镇咳平喘作用[J].承德医学院学报,2001,18(4):277.
[16]左金梁.白屈菜药材及其制剂的质量控制方法研究[D].沈阳:沈阳药科大学.2008.
[17]宗永立,刘艳平.白屈菜红碱对人胃癌BGC823细胞的增殖抑制和凋亡诱导作用[J].中草药.2006.7,37(7):1054-1056.
[18]Lanvers-Kaminsky C,Nolting DM,K(o|¨)ster J.et.al,Anticancer Drugs.17(2006)1025-1030.
[19]Vavreckova C,Gawlik I,M(u|¨)ller K,Benzophenanthridine alkaloids of Chelidonium majus;Ⅱ.Potent inhibitory action against the growth of human keratinocytes[J].Planta Med.1996 Dec;62(6):491-494.
[20]宋立人,洪询,丁绪亮,减载阳.现代中药学大辞典[M].北京:人民卫生出版社,2001:697.
[21]王本样.现代中药药理与临床[M].天津:天津科技翻译出版公司,2004:522.
[22]杨瑞林,魏学智,毕润成.山西霍山罂粟科植物花粉形态[J].广西植物,2003,23(3):231-232
[23]Walterova D,Preininger V,Sirnanek V.Qualitative and quantitative isotaehophoretic analysis of some quaternary isoquinoline alkaloids[J].Planata Med,1984,50(2):149.
[24]Dzido T,Jusiak L,Soezewinski E.Analysis of alkaloid extraet of Chelidonium majus by TLC and micro-bore HPLC[J].Chem Anal,1986,31(1):135.
[25]Bugatti,C.,Colombo,M.L.,Tome,F.High-performance liquid chromatographic separation of quaternary alkaloids of Chelidonium majus L.roots[J].J.Chromatogr.1987,393,312-316.
[26]Han,L.F.,Nowicky,W.,Gutmann,V.,Reversed-phase high-performance liquid chromatographic separation of tertiary and quatemary alkaloids from Chelidoaium majus L[J].J.Chromatogr.1991,543,123-128.
[27]牛长群,何丽一.白屈菜中异哇琳生物碱的高效薄层分离及荧光扫描法[J].药学学报,1991,27(1):69.
[28]Taborska,E.,Bochorakova,H.,Paulova,H.,Dostal,Separation of Alkaloids in Chelidonium majus by Reversed Phase HPLC[J].J.,Planta Med.1994,60,380-381.
[29]Chang-qun,Li-yi He,Determination of isoquinoline alkaloids in Chelidonium majus L.by ion-pair high-performance liquid chromatography[J].J Chromatogr.1991,542,193-199.
[30]Rey,J.P.,Levesque,J.,Pousset,J.L.,Analytical studies of dl-stylopine in Chelidonium majus L.using high-performance liquid chromatography[J].J.Chromatogr.1993,641,180-183.
[31]Kursinszki,L.,Sark(o|¨)zi,A,Kery,A,Sz(o|¨)ke.E,Improved RP-HPLC Method for Analysis of Isoquinoline Alkaloids in Extracts of Chelidonium majus[J].J.Chromatogr,2006,63,S131-S135.
[32]Sark(o|¨)zi,A.,Janicsak,G.,Kursinszki L.,Kery.A.Alkaloid Composition of Chelidonium majus L.Studied by Different Chromatographic Techniques[J].J.Chromatogr,2006,63,S81-S86.
[33]Jana Suchomelova,Hana Bochoiakova,Hana Paulova,Pavel Musil,Eva Tatborska,HPLC quantification of seven quaternary benzo[c]phenanthridine alkaloids in six species of the family Papaveraceae[J].J.Pharm.Biomed.Anal.44(2007) 283-287.
[34]左金梁,白璐,宋欣鑫,等.RP-HPLC法同时测定白屈菜药材中血根碱、小檗碱和白屈菜红碱的含量[J].药物分析杂志.2008,28(6):903-905
[35]曲彩红,陶玲,黄际薇,等.高效液相色谱法测定白屈菜注射液中白屈菜碱的含量[J].中国医院药学杂志.2005,25(7):623-625
[36]韩海.高效液相色谱法同时测定胃痛舒胶囊中原阿片碱和白屈菜碱含量[J].时珍国医国药.2005,16(11):1092-1093
[37]陶玲,曲彩红,黄际薇,等.白屈菜注射液HPLC特征图谱研究[J].中药材,2004.5,27(5):376-377
[38]李晓蒙,朱盛华,廖华卫.白屈菜药材HPLC特征图谱研究[J].天然产物研究与开发.2002.6(14):33
[39]Bo Shao,Hong-zhu Guo,Ya-jun Cui,Ai-hua Liu,Hai-lan Yu,Hui Guo,Man Xu,De-an Guo,Simultaneous determination of six major stilbenes and flavonoids in Smilax china by high performance liquid chromatography[J].J.Pharm.Biomed.Anal.44(2007) 737-742.
[40]Xiao-jie Tan,Qing Li,Xiao-hui Chen,Zhi-wei Wang,Zheng-yuan Shi,Kai-shun Bi,Ying Jia,Simultaneous determination of 13 bioactive compounds in Herba Artemisiae Scopariae(Yin Chen) from different harvest seasons by HPLC-DAD[J].J.Pharm.Biomed.Anal.47(2008) 847-853.
[41]Yang Zhao,Zhangwan Li,Xin Zhou,Zongwei Cai,Xiaojian Gong,Chanyuan Zhou.Quality evaluation of Evodia rutaecarpa(Juss.) Benth by high performance liquid chromatography with photodiode-array detection[J].J.Pharm.Biomed.Anal.48(2008)1230-1236.
[42]Mei Liu,Yongguo Li,Guixin Chou,Xuemei Cheng,Mian Zhang,Zhengtao Wang.Extraction and ultra-performance liquid chromatography of hydrophilic and lipophilic bioactive components in a Chinese herb Radix Salviae Miltiorrhizae[J].J.Chromatogr.A 1157(2007) 51-55.
[43]Li Yang,Aizhen Xiong,Yuqi He,Zaiyong Wang,Changhong Wang,Zhengtao Wang,et.al.Bile Acids Metabonomic Study on the CC_(14)- and a-Naphthylisothiocyanate-Induced Animal Models: Quantitative Analysis of 22 Bile Acids by Ultraperformance Liquid Chromatography-Mass Spectrometry[J]. Chem. Res. Toxicol. 21 (2008) 2280-2288.
[44]J. Guan, CM. Lai, S.P. Li, A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography[J]. J. Pharm. Biomed. Anal. 44 (2007) 996-1000.
[45]Szabolcs Fekete, Jeno Fekete, Katalin Ganzler. Validated UPLC method for the fast and sensitive determination of steroid residues in support of cleaning validation in formulation area[J]. J. Pharm. Biomed. Anal. 49 (2009) 833-838.
[1]傅泉炎.中药含水量与中药仓储质量的关系[J].传统医药.2002,11(5):70.
[2]王淑慧.加强中药材灰分检测[J].现代中药研究与实践.1997.11(3):44.
[3]徐晓光,王宏,董晓辉.中华人民共和国国家标准《人参加工产品分等质量标准》(三)对人参中浸出物含量标准及卫生学标准的研究与讨论[J].人参研究.1995.(2):33.
[4]严辉,段金廒,钱大玮,宿树兰,宋秉生,何子清.我国不同产地当归药材质量的分析与评价[J].中草药.2009.12,40(12):1988-1992.
[5]孙振球.医学统计学(第二版)[M].北京:人民卫生出版社.2006.
[6]Matile.Philipe.Loealization of alkaloids and mechanism of their accumulation in vacuoles of Cheidonium majus laticifers.Nova Acta Leopold Suppl,1976.7:139
[7]冯瑞芝,连文琰,傅桂香,等.罂粟科白屈菜族的化学分类及资源利用[J].植物分类学报.1985,23(1):36.
[8]Slavik J,Slavikova L.Minor alkaloids from Cheldonum majus L[J].Collection Czechoslov Chem Commun,1977,42:2686-2693.
[9]李亚伟,方起程.籽纹紫堇中的异喹啉类生物碱[J].中草药,1991,22(11):486.
[10]王恒山,杨海荣.迭裂黄堇生物碱的研究[J].天然产物研究与开发.1997,(1):37-39.
[11]杜方麓,陈胜璜,阳长明,等.血水草化学成分研究[J].中草药.1993,24(4):177.
[12]于敏,陈卫红,焦连庆.等.白屈菜的研究进展[J].特产研究.2008(2):76-78.
[13]李立军,再帕尔·阿不力孜,项赟,等.黄连总碱的MS/MS及LC-MS/MS研究[J].质谱学报.21(3,4):81-82.
[14]匡海学,董小萍,石任兵.中药化学(第一版).北京:中国中医药出版社.2003:341.