采用UPLC-Q-TOF-MS/MS分析半夏药材中的化学成分
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摘要
目的:采用超高效液相色谱串联四极杆飞行时间高分辨率质谱技术(UPLC-Q-TOF-MS/MS)对半夏的化学成分进行定性分析。方法:采用shim-pack xR-ODSⅢ色谱柱(2. 1 mm×75 mm,1. 6μm),以乙腈-0. 1%甲酸水为流动相进行梯度洗脱,柱温40℃,流速0. 25 m L·min-1,采用电喷雾离子化源,正离子全扫描的一级和二级质谱信息。结果:通过一级精确荷质比和二级的碎片数据、参考文献或数据库,质谱裂解规律和标准品的保留时间,发现90个化学成分,初步定性80个化学成分,分别为7个生物碱类,8个醇醚类,12个脂肪酸甘油脂类,5个黄酮类,12个溶血磷脂酰胆碱类,10个醇氨类,11个氨基酸类,11个酰胺类,其他类型4个化合物。结论:通过UPLC-Q-TOF-MS/MS对半夏化学成分时行分析,发现半夏中含有溶血磷脂酰胆碱类物质,其可引起炎症反应及神经元髓鞘脱失和不同程度变性。可推测其可能与半夏的刺激性毒性相关,同时对半夏化学成分全面分析也为半夏药效物质基础研究和质量控制奠定了一定的基础。
Objective: To identify the chemical constituents of Pinelliae Rhizoma( BX) by ultrahighpressure liquid chromatography coupled with electrospray time-of-flight tandem mass spectrometry( UPLC-Q-TOFMS/MS). Method: Chromatograpic separation was performed on a shim-pack xR-ODS Ⅲ column( 2. 1 mm ×75 mm,1. 6 μm) using a gradient elution program with mixtures of acetonitrile and 0. 1% formic acid-water as mobile phases at a flow rate of 0. 25 m L·min-1. UPLC-Q-TOF-MS/MS was developed for rapid and highthroughput screening of the preliminary chemical profile of BX in positive ion modes. Result: Ninety chemical components were found in BX,according to the accurate Charge-mass Ratio and the MS/MS data,retention time of reference standard, references or databases, the fragmentation regularities of mass spectra. Eighty were identified preliminarily, including 7 alkaloids, 8 poly-alcohols, 12 fatty glycerides, 5 flavonoids, 12 lysophosphatidylcholines( LPCs),10 alcohol amines,11 amino acids,11 amides and 4 other type. Conclusion:LPCs were first found in BX. Because BX has certain toxicity,it is found that BX contains LPCs by analyzing the chemical constituents of BX, which can cause inflammatory reaction and neuronal myelin sheath loss and degeneration. Therefore,the analysis of the chemical composition of BX can explain the causes of the toxicity and provide a foundation for the basic research and quality control of the potency of BX.
Objective: To identify the chemical constituents of Pinelliae Rhizoma( BX) by ultrahighpressure liquid chromatography coupled with electrospray time-of-flight tandem mass spectrometry( UPLC-Q-TOFMS/MS). Method: Chromatograpic separation was performed on a shim-pack xR-ODS Ⅲ column( 2. 1 mm ×75 mm,1. 6 μm) using a gradient elution program with mixtures of acetonitrile and 0. 1% formic acid-water as mobile phases at a flow rate of 0. 25 m L·min-1. UPLC-Q-TOF-MS/MS was developed for rapid and highthroughput screening of the preliminary chemical profile of BX in positive ion modes. Result: Ninety chemical components were found in BX,according to the accurate Charge-mass Ratio and the MS/MS data,retention time of reference standard, references or databases, the fragmentation regularities of mass spectra. Eighty were identified preliminarily, including 7 alkaloids, 8 poly-alcohols, 12 fatty glycerides, 5 flavonoids, 12 lysophosphatidylcholines( LPCs),10 alcohol amines,11 amino acids,11 amides and 4 other type. Conclusion:LPCs were first found in BX. Because BX has certain toxicity,it is found that BX contains LPCs by analyzing the chemical constituents of BX, which can cause inflammatory reaction and neuronal myelin sheath loss and degeneration. Therefore,the analysis of the chemical composition of BX can explain the causes of the toxicity and provide a foundation for the basic research and quality control of the potency of BX.
引文
[1]张亚囡,黄幼异,鲍志烨,等.半夏酸水渗漉液单次给药对小鼠肝毒性“量-时-毒”关系研究[J].中国药物警戒,2011,8(1):15-19.
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[15] Maruno M. Active pliciples of pinelliae tuder and new preparation of crude drug[J]. Wakan Lyakugaku Zasshi,1997,14(2):81-88.
[16] CHEN J H,CUI G Y,LIU J Y,et al. Pinelloside,an antimicrobial cerebroside from Pinellia ternata[J].Phytochemistry,2003,64(4):903-906.
[17]陈金梅,廖锦红,高金薇,等. UPLC-Q-TOF-MS/MS研究胆木药材水提物的化学成分[J].中国实验方剂学杂志,2018,24(15):109-116.
[18]何芋岐,鲁艳柳,李利生,等.基于UPLC-ESI-OrbitrapMS技术对金钗石斛生物碱的分析[J].中国实验方剂学杂志,2017,23(20):30-35.
[19]李玲云,王云,刘梦娇,等.栀子金花丸化学成分的UPLC-Q-TOF-MS/MS快速鉴定与分析[J].中国实验方剂学杂志,2017,23(14):1-11.
[20] Gosettia F,Chiuminatto U,Mazzucco E,et al. Ultra-highperformance liquid chromatography/tandem highresolution mass spectrometry analysis of sixteen red beverages containing carminic acid:identification of degradation products by using principal component analysis/discriminant analysis[J]. Food Chem,2015,167:454-462.
[21]李定祥,王珍,罗建光,等.坤泰胶囊化学成分的LCESI-MS/MS分析[J].中国实验方剂学杂志,2017,23(19):90-93.
[22] Chan E C,Yap S L,Lau A J,et al. Ultra-performance liquid chromatography/time-of-flight mass spectrometry based metabolomics of raw and steamed Panax notoginseng[J]. Rapid Comm Mass Spectrom,2007,21(4):519-528.
[23] Pedro A,Eva L,YANG Y,et al. The impact of exogenousω'-6 andω'-3 polyunsaturated fatty acids on the induced production of pro-and anti-inflammatory prostaglandins and leukotrienes in Atlantic salmon head kidney cells using a full factorial design and LC-MS/MS[J]. J Chromatogra B,2014,964:164-171.
[24] ZHANG Z H,ZHAO Y Y,CHENG X L,et al. General toxicity of Pinellia ternate(Thunb.)Berit. in rat:A metabonomic method for profiling of serum metabolic changes[J]. J Ethnopharmacol,2013,149(1):303-310.
[25] Seefelder S,Schwerdt G,Freudinger R,et al. Liquid chromatography/electrospray ionisation-mass spectrometry method for the quantification of sphingosine and sphinganine in cell cultures exposed to fumonisins[J]. J Chromatogr B,2002,780(1):137-144.
[26]吴世芳.何首乌中磷脂类化合物的提取、分离及分析方法研究[D].南昌:南昌大学,2007.
[27] GENG L,SUN H,YUAN Y,et al. Discrimination of raw and vinegar-processed Genkwa Flos using metabolomics coupled with multivariate data analysis:a discrimination study with metabolomics coupled with PCA[J].Fitoterapia,2013,84(1):286-294.
[28]亢爱春,霍勇,齐丽彤.溶血磷脂酰胆碱在动脉粥样硬化中的作用[J].中国动脉硬化杂志,2006,14(12):1083-1086.
[29]刘小丰,钱祝银,苗毅.溶血卵磷脂在胰性脑病发病机制中的作用[J].中华实验外科杂志,2005,22(4):413-415.
[2]杨守业,叶文华,吴子伦,等.半夏炮制前后对小白鼠急性、亚急性和蓄积性毒性的研究[J].中成药,1988(7):18-19.
[3] ZHANG Z H,ZHAO Y Y,CHENG X L,et al. General toxicity of Pinellia ternate(Thunb.)Berit. in rat:A metabonomic method for profiling of serum metabolic changes[J]. J Ethnopharmacol,2013,149(1):303-310.
[4] Shin S,Park D,Jeon J H,et al. Effect of Pinellia ternata,extract on fetal development of rats[J].Reproduct Toxicol,2007,24(1):74.
[5] ZHANG Z H,ZHAO Y Y,CHENG X L,et al.Metabonomic study of biochemical changes in the rat urine induced by Pinellia ternata(Thunb.)Berit.[J].J Pharm Biomed Anal,2013,85(1):186-193.
[6] LI S L,SONG J Z,QIAO C F,et al. A novel strategy to rapidly explore potential chemical markers for the discrimination between raw and processed Radix Rehmanniae by UHPLC-TOFMS with multivariate statistical analysis[J]. J Pharm Biomed Anal,2010,51(4):812-823.
[7]魏运姣.法半夏炮制工艺及质量研究[D].武汉:湖北中医学院,2005.
[8]丸野正雄. Active priciples of pinelliae tube and new preparation of crude drug[J]. J Traditional Med,1997,14(2):81-82.
[9]张科卫,吴皓,武露凌.半夏药材中脂肪酸成分的研究[J].南京中医药大学学报:自然科学版,2002,18(5):291-292.
[10]吴皓,张科卫,李伟,等.半夏的化学成分研究[J].中草药,2003,34(7):593-594.
[11]大盐春治. Isolation of L-ephedrine from “pinelliae tuber” chemical[J]. Pharmaceut Bull,1978,26(7):2096.
[12]范美华,周吉源.半夏的研究进展[J].西北药学杂志,2004,19(2):90-92.
[13]王锐,倪京满,马蓉,等.中药半夏挥发油成分的研究[J].中国药学杂志,1995,30(8):457-459.
[14]李先端,胡世林,杨连菊.半夏类药材氨基酸与无机元素分析[J].中国中药杂志,1990,15(10):37-38.
[15] Maruno M. Active pliciples of pinelliae tuder and new preparation of crude drug[J]. Wakan Lyakugaku Zasshi,1997,14(2):81-88.
[16] CHEN J H,CUI G Y,LIU J Y,et al. Pinelloside,an antimicrobial cerebroside from Pinellia ternata[J].Phytochemistry,2003,64(4):903-906.
[17]陈金梅,廖锦红,高金薇,等. UPLC-Q-TOF-MS/MS研究胆木药材水提物的化学成分[J].中国实验方剂学杂志,2018,24(15):109-116.
[18]何芋岐,鲁艳柳,李利生,等.基于UPLC-ESI-OrbitrapMS技术对金钗石斛生物碱的分析[J].中国实验方剂学杂志,2017,23(20):30-35.
[19]李玲云,王云,刘梦娇,等.栀子金花丸化学成分的UPLC-Q-TOF-MS/MS快速鉴定与分析[J].中国实验方剂学杂志,2017,23(14):1-11.
[20] Gosettia F,Chiuminatto U,Mazzucco E,et al. Ultra-highperformance liquid chromatography/tandem highresolution mass spectrometry analysis of sixteen red beverages containing carminic acid:identification of degradation products by using principal component analysis/discriminant analysis[J]. Food Chem,2015,167:454-462.
[21]李定祥,王珍,罗建光,等.坤泰胶囊化学成分的LCESI-MS/MS分析[J].中国实验方剂学杂志,2017,23(19):90-93.
[22] Chan E C,Yap S L,Lau A J,et al. Ultra-performance liquid chromatography/time-of-flight mass spectrometry based metabolomics of raw and steamed Panax notoginseng[J]. Rapid Comm Mass Spectrom,2007,21(4):519-528.
[23] Pedro A,Eva L,YANG Y,et al. The impact of exogenousω'-6 andω'-3 polyunsaturated fatty acids on the induced production of pro-and anti-inflammatory prostaglandins and leukotrienes in Atlantic salmon head kidney cells using a full factorial design and LC-MS/MS[J]. J Chromatogra B,2014,964:164-171.
[24] ZHANG Z H,ZHAO Y Y,CHENG X L,et al. General toxicity of Pinellia ternate(Thunb.)Berit. in rat:A metabonomic method for profiling of serum metabolic changes[J]. J Ethnopharmacol,2013,149(1):303-310.
[25] Seefelder S,Schwerdt G,Freudinger R,et al. Liquid chromatography/electrospray ionisation-mass spectrometry method for the quantification of sphingosine and sphinganine in cell cultures exposed to fumonisins[J]. J Chromatogr B,2002,780(1):137-144.
[26]吴世芳.何首乌中磷脂类化合物的提取、分离及分析方法研究[D].南昌:南昌大学,2007.
[27] GENG L,SUN H,YUAN Y,et al. Discrimination of raw and vinegar-processed Genkwa Flos using metabolomics coupled with multivariate data analysis:a discrimination study with metabolomics coupled with PCA[J].Fitoterapia,2013,84(1):286-294.
[28]亢爱春,霍勇,齐丽彤.溶血磷脂酰胆碱在动脉粥样硬化中的作用[J].中国动脉硬化杂志,2006,14(12):1083-1086.
[29]刘小丰,钱祝银,苗毅.溶血卵磷脂在胰性脑病发病机制中的作用[J].中华实验外科杂志,2005,22(4):413-415.