黄连多糖中单糖组成的HPLC-MS~n法快速识别
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摘要
目的:探讨采用HPLC-MS~n法快速识别黄连多糖中的单糖组成。方法:黄连药材经过提取、分离、除蛋白、除色素,最终得到黄连多糖,用2 mol/L的三氟乙酸水解黄连多糖,以1-苯基-3-甲基-5-吡唑啉酮(PMP)作单糖衍生化试剂,利用HPLC-MS~n识别黄连中单糖的组成,并解析其衍生物的裂解规律。结果:通过分析,得到的黄连多糖中含有甘露糖、核糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖/木糖、岩藻糖,其中核糖、岩藻糖为首次在黄连多糖中报道的单糖。结论:HPLC-MS~n法对中药多糖组分中的单糖成分进行分析具有重要的指导意义。
Objective: To explore a rapid identification of monosaccharide composition of polysaccharide from Rhizoma Coptidis through HPLC-MS~n. Methods: After being extracted,separated,removed of protein and pigment,Rhizoma Coptidis polysaccharide was obtained from Rhizoma Coptidis. Polysaccharides of Rhizoma Coptidis was hydrolyzed with the TFA of 2 mol. L~(-1). 1-phenyl-3-methyl-5-pyrazolone( PMP) was used as derivatization reagent. And high performance liquid chromatography-mass spectrometry( HPLC-MS~n) was used to identify the composition of monosaccharide,and the resolve fragmentation regularity of its derivatives was analyzed. Results: Through analysis,the monosaccharides of polysaccharides from Rhizoma Coptidis were mannose,ribose,rhamnose,galacturonic acid,glucose,galactose,arabinose/xylose,fucose,among which ribose and fucose were the first reported monosaccharides that obtained from polysaccharide of Rhizoma Coptidis. Conclusion: HPLC-MS~nhas a guiding significance in analyzing the monosaccharide composition of polysaccharides from Chinese materia medica.
Objective: To explore a rapid identification of monosaccharide composition of polysaccharide from Rhizoma Coptidis through HPLC-MS~n. Methods: After being extracted,separated,removed of protein and pigment,Rhizoma Coptidis polysaccharide was obtained from Rhizoma Coptidis. Polysaccharides of Rhizoma Coptidis was hydrolyzed with the TFA of 2 mol. L~(-1). 1-phenyl-3-methyl-5-pyrazolone( PMP) was used as derivatization reagent. And high performance liquid chromatography-mass spectrometry( HPLC-MS~n) was used to identify the composition of monosaccharide,and the resolve fragmentation regularity of its derivatives was analyzed. Results: Through analysis,the monosaccharides of polysaccharides from Rhizoma Coptidis were mannose,ribose,rhamnose,galacturonic acid,glucose,galactose,arabinose/xylose,fucose,among which ribose and fucose were the first reported monosaccharides that obtained from polysaccharide of Rhizoma Coptidis. Conclusion: HPLC-MS~nhas a guiding significance in analyzing the monosaccharide composition of polysaccharides from Chinese materia medica.
引文
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[3]王舒.中药黄连研究进展[J].亚太传统医药,2015,11(9):44-46.
[4]张月月,王君明,崔瑛,等.中药生物碱类成分抗焦虑或抑郁活性及黄连生物碱药理作用研究进展[J].中华中医药杂志,2015,30(4):1184-1187.
[5]鲁周南,包晓霞,薛晓鸥,等.黄连抗肿瘤临床运用及安全性评估研究进展[J].辽宁中医药大学学报,2017,19(1):131-133.
[6]陆瑞群,庞雅琴,庞广福.岩黄连的药理作用及临床应用研究进展[J].中国医药导报,2014,11(5):159-160,165.
[7]徐维相.探讨中药黄连的研究进展[J].世界最新医学信息文摘(连续型电子期刊),2016,16(22):13,12.
[8]尹丽丽.黄连的药理作用及现代研究进展[J].中医临床研究,2016,8(28):144-145.
[9]曲华,张莹,冒慧敏,等.黄连及其提取物治疗心血管疾病的临床研究进展[J].中医药导报,2016,22(6):103-106.
[10]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RPHPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,42(24):4579-4586.
[11]王婷婷,钟凌云.姜制黄连炮制近年来研究进展[J].时珍国医国药,2016,27(2):427-429.
[12]卿大双,罗维早,孙建彬,等.一测多评法测定黄连及其炮制品中6种生物碱[J].中草药,2016,42(2):324-329.
[13]刘丹,曹广尚,司席席,等.黄连中生物碱类成分抗心律失常研究概述[J].山东中医杂志,2017,36(2):164-166,171.
[14]黄程程,孙磊,张泽辉,等.中药有效成分对细菌溶血素的抑制作用[J].中国畜牧兽医,2016,43(12):3363-3367.
[15]王利红,唐文照,辛义周.黄连中生物碱成分及药理作用研究进展[J].山东中医药大学学报,2015,39(4):389-392.
[16]刘卫霞,尤凯,孙燕.黄连解毒汤化学成分及临床应用的研究进展[J].沈阳医学院学报,2015,17(2):121-124.
[17]常惠,吴效科.黄连素对PCOS治疗作用及相关机制研究进展[J].现代妇产科进展,2017,26(3):235-237.
[18]马聪颖,陈佳纬,卢长林.黄连素在心血管疾病应用的研究进展[J].中西医结合心血管病(连续型电子期刊),2016,4(31):37-39.
[19]王维琪.黄连有效成分的药理研究与进展[J].中医临床研究,2016,8(26):147-148.
[20]张青,李琰,陈磊.黄连素对2型糖尿病及其并发症的治疗作用及相关机制研究进展[J].中国中药杂志,2015,40(9):1660-1665.
[21]陈瑞,吴云,田维毅.黄连化学成分及其影响相关代谢途径的研究进展[J].湖南中医杂志,2016,32(4):190-192.
[22]孟芳,谭小武.黄连素诱导肿瘤细胞凋亡机制的研究进展[J].中国现代医药杂志,2016,18(4):99-102.
[23]孙宁阳,张宏川,顾健,等.葛根黄连组分毒理学研究[J].中药药理与临床,2017,33(1):95-98.
[24]Fan G,Tao LH,Yue QH,et al.Metabolic discrimination of rhizoma coptidis from different species using 1H NMR spectroscopy and principal component analysis[J].Planta Med,2012,78(6):641-648.
[25]Fan G,Zhang MY,Zhou XD,et al.Quality evaluation and species differentiation of Rhizoma coptidis by using proton nuclear magnetic resonance spectroscopy[J].Anal Chim Acta,2012,747:76-83.
[26]霍志鹏,王玉,周水平,等.黄连生物碱成分的HPLCDADMS分析[J].天津药学,2016,28(2):8-12.
[27]黄涛阳,王晖,翁燕君,等.电喷雾电离-串联质谱快速分析黄连解毒汤的化学成分[J].中药材,2017,40(1):119-121.
[28]李云,王炜,尹登科,等.黄连多糖不同组分抗氧化活性比较研究[J].安徽中医药大学学报,2015,34(1):66-69.
[29]李敏,金晶,叶新,等.黄连多糖抗蛋白质非酶糖基化活性初步研究[J].黑龙江大学自然科学学报,2014,31(2):223-227.
[30]姜爽,李璐,王星云,等.黄连多糖提取工艺的优化及其体外抗氧化作用[J].中成药,2016,38(6):1405-1407.
[31]王伟,刘世军,安法娥,等.黄连多糖提取、分离纯化鉴定及清除羟基自由基能力测定[J].山东医药,2013,53(10):79-82.
[32]尹登科,杨晔,陈松,等.黄连多糖对AGEs诱导内皮细胞增殖及其受体表达的作用研究[J].药物生物技术,2012,19(6):476-479.
[33]Honda S,Akao E,Suzuki S,et al.High-performance liquid chromatography of reducing carbohydrates as strongly ultraviolet-absorbing and electrochemically sensitive 1-phenyl-3-methyl-5-pyrazolone derivatives[J].Anal Biochem,1989,180(2):351-357.
[34]Wang HX,Zhao J,Li DM,et al.Structural investigation of a uronic acid-containing polysaccharide from abalone by graded acid hydrolysis followed by PMP-HPLC-MSnand NMR analysis[J].Carbohydr Res,2015,402:95-101.
[35]Wu X,Jiang W,Lu J,et al.Analysis of the monosaccharide composition of water-soluble polysaccharides from Sargassum fusiforme by high performance liquid chromatography/electrospray ionisation mass spectrometry[J].Food Chem,2014,145:976-983.
[36]Wang H,Zhao J,Li D,et al.Comparison of polysaccharides of Haliotis discus hannai and Volutharpa ampullacea perryi by PMP-HPLCMS(n)analysis upon acid hydrolysis[J].Carbohydr Res,2015,415:48-53.
[37]赵宏,柴桂芳,王秋红,等.一种车前子多糖的分离纯化及单糖组成分析[J].中国实验方剂学杂志,2014,20(19):97-100.
[38]吕永磊,卜海博,杨蕾,等.乌头母根、子根、须根多糖的比较[J].中国中药杂志,2011,36(9):1154-1157.
[2]王强.中药分析[M].北京:中国医药科技出版社,2005:306.
[3]王舒.中药黄连研究进展[J].亚太传统医药,2015,11(9):44-46.
[4]张月月,王君明,崔瑛,等.中药生物碱类成分抗焦虑或抑郁活性及黄连生物碱药理作用研究进展[J].中华中医药杂志,2015,30(4):1184-1187.
[5]鲁周南,包晓霞,薛晓鸥,等.黄连抗肿瘤临床运用及安全性评估研究进展[J].辽宁中医药大学学报,2017,19(1):131-133.
[6]陆瑞群,庞雅琴,庞广福.岩黄连的药理作用及临床应用研究进展[J].中国医药导报,2014,11(5):159-160,165.
[7]徐维相.探讨中药黄连的研究进展[J].世界最新医学信息文摘(连续型电子期刊),2016,16(22):13,12.
[8]尹丽丽.黄连的药理作用及现代研究进展[J].中医临床研究,2016,8(28):144-145.
[9]曲华,张莹,冒慧敏,等.黄连及其提取物治疗心血管疾病的临床研究进展[J].中医药导报,2016,22(6):103-106.
[10]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RPHPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,42(24):4579-4586.
[11]王婷婷,钟凌云.姜制黄连炮制近年来研究进展[J].时珍国医国药,2016,27(2):427-429.
[12]卿大双,罗维早,孙建彬,等.一测多评法测定黄连及其炮制品中6种生物碱[J].中草药,2016,42(2):324-329.
[13]刘丹,曹广尚,司席席,等.黄连中生物碱类成分抗心律失常研究概述[J].山东中医杂志,2017,36(2):164-166,171.
[14]黄程程,孙磊,张泽辉,等.中药有效成分对细菌溶血素的抑制作用[J].中国畜牧兽医,2016,43(12):3363-3367.
[15]王利红,唐文照,辛义周.黄连中生物碱成分及药理作用研究进展[J].山东中医药大学学报,2015,39(4):389-392.
[16]刘卫霞,尤凯,孙燕.黄连解毒汤化学成分及临床应用的研究进展[J].沈阳医学院学报,2015,17(2):121-124.
[17]常惠,吴效科.黄连素对PCOS治疗作用及相关机制研究进展[J].现代妇产科进展,2017,26(3):235-237.
[18]马聪颖,陈佳纬,卢长林.黄连素在心血管疾病应用的研究进展[J].中西医结合心血管病(连续型电子期刊),2016,4(31):37-39.
[19]王维琪.黄连有效成分的药理研究与进展[J].中医临床研究,2016,8(26):147-148.
[20]张青,李琰,陈磊.黄连素对2型糖尿病及其并发症的治疗作用及相关机制研究进展[J].中国中药杂志,2015,40(9):1660-1665.
[21]陈瑞,吴云,田维毅.黄连化学成分及其影响相关代谢途径的研究进展[J].湖南中医杂志,2016,32(4):190-192.
[22]孟芳,谭小武.黄连素诱导肿瘤细胞凋亡机制的研究进展[J].中国现代医药杂志,2016,18(4):99-102.
[23]孙宁阳,张宏川,顾健,等.葛根黄连组分毒理学研究[J].中药药理与临床,2017,33(1):95-98.
[24]Fan G,Tao LH,Yue QH,et al.Metabolic discrimination of rhizoma coptidis from different species using 1H NMR spectroscopy and principal component analysis[J].Planta Med,2012,78(6):641-648.
[25]Fan G,Zhang MY,Zhou XD,et al.Quality evaluation and species differentiation of Rhizoma coptidis by using proton nuclear magnetic resonance spectroscopy[J].Anal Chim Acta,2012,747:76-83.
[26]霍志鹏,王玉,周水平,等.黄连生物碱成分的HPLCDADMS分析[J].天津药学,2016,28(2):8-12.
[27]黄涛阳,王晖,翁燕君,等.电喷雾电离-串联质谱快速分析黄连解毒汤的化学成分[J].中药材,2017,40(1):119-121.
[28]李云,王炜,尹登科,等.黄连多糖不同组分抗氧化活性比较研究[J].安徽中医药大学学报,2015,34(1):66-69.
[29]李敏,金晶,叶新,等.黄连多糖抗蛋白质非酶糖基化活性初步研究[J].黑龙江大学自然科学学报,2014,31(2):223-227.
[30]姜爽,李璐,王星云,等.黄连多糖提取工艺的优化及其体外抗氧化作用[J].中成药,2016,38(6):1405-1407.
[31]王伟,刘世军,安法娥,等.黄连多糖提取、分离纯化鉴定及清除羟基自由基能力测定[J].山东医药,2013,53(10):79-82.
[32]尹登科,杨晔,陈松,等.黄连多糖对AGEs诱导内皮细胞增殖及其受体表达的作用研究[J].药物生物技术,2012,19(6):476-479.
[33]Honda S,Akao E,Suzuki S,et al.High-performance liquid chromatography of reducing carbohydrates as strongly ultraviolet-absorbing and electrochemically sensitive 1-phenyl-3-methyl-5-pyrazolone derivatives[J].Anal Biochem,1989,180(2):351-357.
[34]Wang HX,Zhao J,Li DM,et al.Structural investigation of a uronic acid-containing polysaccharide from abalone by graded acid hydrolysis followed by PMP-HPLC-MSnand NMR analysis[J].Carbohydr Res,2015,402:95-101.
[35]Wu X,Jiang W,Lu J,et al.Analysis of the monosaccharide composition of water-soluble polysaccharides from Sargassum fusiforme by high performance liquid chromatography/electrospray ionisation mass spectrometry[J].Food Chem,2014,145:976-983.
[36]Wang H,Zhao J,Li D,et al.Comparison of polysaccharides of Haliotis discus hannai and Volutharpa ampullacea perryi by PMP-HPLCMS(n)analysis upon acid hydrolysis[J].Carbohydr Res,2015,415:48-53.
[37]赵宏,柴桂芳,王秋红,等.一种车前子多糖的分离纯化及单糖组成分析[J].中国实验方剂学杂志,2014,20(19):97-100.
[38]吕永磊,卜海博,杨蕾,等.乌头母根、子根、须根多糖的比较[J].中国中药杂志,2011,36(9):1154-1157.