UHPLC-Q-TOF MS和化学计量学方法分析川芎硫磺熏蒸前后化学成分的变化
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摘要
采用UHPLC-Q-TOF MS结合化学计量学方法分析川芎硫磺熏蒸前后化学成分的变化。选取19批未经硫磺熏蒸川芎饮片、11批实验室自制硫磺熏蒸川芎饮片和9批市售硫磺熏蒸川芎饮片作为实验样品,制备供试品溶液进样后,应用PeakView1.2软件依据一级质谱的精确质荷比和二级质谱的碎片信息进行成分解析,并结合MarkerView1.2.1软件进行主成分分析(PCA)和t检验,鉴定其中的共有成分和差异性成分。结果表明,在PCA分析中,正负离子模式下的未硫熏和硫熏川芎样品均可被明显地区分开,主成分之间存在明显的差异;t检验结果也显示二者之间存在大量的差异性成分,同时鉴定出部分差异性成分,并进一步推测了其可能的转化方式。本实验可为研究硫磺熏蒸对川芎质量的影响提供客观参考,并为控制硫磺熏蒸在中药材和饮片加工过程中的过度应用和确保中医临床用药的安全提供理论依据。
Sulfur-fumigation has been used around the world for centuries in the food industry to inhibit oxidation of the light-colored fruits or vegetables,and this sulfurbased preservative approach has also been employed in postharvest handling of the medicinal herbs to preserve color and freshness,and to protect against insects and molds over the last several decades in China.However,more and more recent reports about sulfur-fumigation show that it can cause the chemical transformations of the original bioactive components(the sulfur dioxide obtained from the heating of sulfur,as a strong reducing agent,may react with the ketonic group and hydroxyl radical-containing components of the medicinal herbs),affect the efficacies and the pharmacokinetics of the medicinal herbs and their extracts or preparations,and more seriously it can even lead to a toxic reaction in the liver and kidneys.Therefore,the influence on the intrinsic quality of Chuanxiong Rhizomacaused by sulfur-fumigation needs to be investigated systematically.In this paper,the differences of chemical constituents between sun-dried and sulfur-fumigated Chuanxiong Rhizoma were investigated based on UHPLC-Q-TOF MS and chemometric analysis.The samples of 19 batches of sun-dried Chuanxiong Rhizoma,11 batches of lab-prepared sulfur-fumigated Chuanxiong Rhizoma,and 9batches of industrial sulfur-fumigated Chuanxiong Rhizoma were taken as the research objects.The sample solutions were prepared by the optimized extraction method.After injection of the prepared sample solutions,PeakView1.2software was applied to analyze the mass spectrometric data and to identify the constituents based on the accurate m/z values of Q-TOF MS and the fragments of Q-TOF MS2.Meanwhile,MarkerView1.2.1software was combined to develop the principal component analysis and t-test statistical analysis to identify as more common components and components with significant differences as possible.The results showed that the samples of sun-dried and sulfur-fumigated Chuanxiong Rhizoma could be classified clearly both in positive and negative ion modes by the means of principal component analysis,and there were obvious statistical differences existed among the principal components.What's more,the results of t-test also demonstrated that there were a large number of constituents with statistical differences existed between sun-dried and sulfur-fumigated Chuanxiong Rhizoma,and parts of them were identified and their possible transformation pathways were further proposed.The study can provide the objective reference for the influence of sulfur-fumigation on the quality of Chuanxiong Rhizoma,and the theoretical basis for the control of excessive use of sulfur-fumigation in the processing of Chinese medicinal herbs and for the ensurance of safety and effectiveness of clinical medication in Chinese medicine.
Sulfur-fumigation has been used around the world for centuries in the food industry to inhibit oxidation of the light-colored fruits or vegetables,and this sulfurbased preservative approach has also been employed in postharvest handling of the medicinal herbs to preserve color and freshness,and to protect against insects and molds over the last several decades in China.However,more and more recent reports about sulfur-fumigation show that it can cause the chemical transformations of the original bioactive components(the sulfur dioxide obtained from the heating of sulfur,as a strong reducing agent,may react with the ketonic group and hydroxyl radical-containing components of the medicinal herbs),affect the efficacies and the pharmacokinetics of the medicinal herbs and their extracts or preparations,and more seriously it can even lead to a toxic reaction in the liver and kidneys.Therefore,the influence on the intrinsic quality of Chuanxiong Rhizomacaused by sulfur-fumigation needs to be investigated systematically.In this paper,the differences of chemical constituents between sun-dried and sulfur-fumigated Chuanxiong Rhizoma were investigated based on UHPLC-Q-TOF MS and chemometric analysis.The samples of 19 batches of sun-dried Chuanxiong Rhizoma,11 batches of lab-prepared sulfur-fumigated Chuanxiong Rhizoma,and 9batches of industrial sulfur-fumigated Chuanxiong Rhizoma were taken as the research objects.The sample solutions were prepared by the optimized extraction method.After injection of the prepared sample solutions,PeakView1.2software was applied to analyze the mass spectrometric data and to identify the constituents based on the accurate m/z values of Q-TOF MS and the fragments of Q-TOF MS2.Meanwhile,MarkerView1.2.1software was combined to develop the principal component analysis and t-test statistical analysis to identify as more common components and components with significant differences as possible.The results showed that the samples of sun-dried and sulfur-fumigated Chuanxiong Rhizoma could be classified clearly both in positive and negative ion modes by the means of principal component analysis,and there were obvious statistical differences existed among the principal components.What's more,the results of t-test also demonstrated that there were a large number of constituents with statistical differences existed between sun-dried and sulfur-fumigated Chuanxiong Rhizoma,and parts of them were identified and their possible transformation pathways were further proposed.The study can provide the objective reference for the influence of sulfur-fumigation on the quality of Chuanxiong Rhizoma,and the theoretical basis for the control of excessive use of sulfur-fumigation in the processing of Chinese medicinal herbs and for the ensurance of safety and effectiveness of clinical medication in Chinese medicine.
引文
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[8]练杭芸,徐王彦君,梁乾德,等.UPLC-QTOF MS法比较赤芍、白芍煎液化学成分差异[J].质谱学报,2014,35(3):269-278.LIAN Hangyun,XU Wangyanjun,LIANG Qiande,et al.Chemical comparison between decoctions of Radix Paeoniae Rubra and Radix Paeoniae Alba by UPLC-QTOF MS[J].Journal of Chinese Mass Spectrometry Society,2014,35(3):269-278(in Chinese).
[9]侯娅,马阳,邹立思,等.基于UPLC-Triple-TOFMS/MS技术分析不同产地太子参的差异化学成分[J].质谱学报,2015,36(4):359-366.HOU Ya,MA Yang,ZOU Lisi,et al.Difference of chemical compositions in Pseudostellariae Radix from different origins by UPLC-Triple TOF-MS/MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(4):359-366(in Chinese).
[10]王静哲,刘震,马立满,等.基于UPLC-Q-TOF MS分析加工炮制对玄参化学成分的影响[J].质谱学报,2016,37(1):1-9.WANG Jingzhe,LIU Zhen,MA Liman,et al.Changes of chemical constituents in Scrophulariae Radix during processing based on UPLC-QTOF MS[J].Journal of Chinese Mass Spectrometry Society,2016,37(1):1-9(in Chinese).
[11]张小平,蒋可志,吕惠卿,等.HPLC-Q-TOF MS鉴定条叶榕根茎乙酸乙酯提取物中的主要化学成分[J].质谱学报,2015,36(4):310-320.ZHANG Xiaoping,JIANG Kezhi,LV Huiqing,et al.Identification and characterization of major chemical compounds in the ethyl acetate extract from Ficus pandurata Hance Aerial Roots by HPLC-Q-TOF MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(4):310-320(in Chinese).
[12]宋燕燕,李敏,吴柱樟,等.采用UPLC-Q-TOF MS技术分析坛紫菜中脂质成分[J].质谱学报,2015,36(5):425-433.SONG Yanyan,LI Min,WU Zhuzhang,et al.Rapid identification of lipid compositions from Porphyra haitanensis by UPLC-Q-TOF MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(5):425-433(in Chinese).
[13]马强,白桦,王超,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查化妆品中18种香豆素类化合物[J].分析测试学报,2014,33(3):248-255.MA Qiang,BAI Hua,WANG Chao,et al.Rapid screening of 18coumarin compounds in cosmetics by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J].Journal of Instrumental Analysis,2014,33(3):248-255(in Chinese).
[14]董静,王弘,万乐人,等.高效液相色谱/电喷雾-离子阱-飞行时间质谱分析鉴定中药虎杖中的主要化学成分[J].色谱,2009,27(4):425-430.DONG Jing,WANG Hong,WAN Leren,et al.Identification and determination of major constituents in Polygonum cuspidatum Sieb.et Zucc.by high performance liquid chromatography/electrospray ionization-ion trap-time-of-flight mass spectrometry[J].Chinese Journal of Chromatography,2009,27(4):425-430(in Chinese).
[2]李松林,林鸽,钟凯声,等.应用HPLC-DAD-MS联用技术研究中药川芎指纹图谱[J].药学学报,2004,39(8):621-626.LI Songlin,LIN Ge,CHUNG Hoising,et al.Study on fingerprint of Rhizome Chuanxiong by HPLC-DAD-MS[J].Acta Pharmaceutica Sinica,2004,39(8):621-626(in Chinese).
[3]彭仕华,周同惠.丁基苯酞的体内代谢转化研究[J].药学学报,1996,31(10):780-784.PENG Shihua,ZHOU Tonghui.Investigation on in vivo metabolism of n-butyl phthalide[J].Acta Pharmaceutica Sinica,1996,31(10):780-784(in Chinese).
[4]徐皓亮,冯亦璞.丁基苯酞对大鼠血栓形成及血小板功能的影响[J].药学学报,2001,36(5):329-333.XU Haoliang,FENG Yipu.Effects of 3-Nbutylphthalide on thrombosis formation and platelet function in rats[J].Acta Pharmaceutica Sinica,2001,36(5):329-333(in Chinese).
[5]刘静静,刘晓,李松林,等.硫磺熏蒸中药材及饮片的研究现状[J].中草药,2010,41(8):1 403-1 406.LIU Jingjing,LIU Xiao,LI Songlin,et al.Current situation in studies on traditional Chinese medicinal materials and Yinpian by sulfur-fumigated process[J].Chinese Traditional and Herbal Drugs,2010,41(8):1 403-1 406(in Chinese).
[6]LI S L,SONG J Z,CHOI F F K,et al.Chemical profiling of Radix Paeoniae evaluated by ultra performance liquid chromatography/photo-diodearray/quadrupole time-of-flight mass spectrometry[J].Journal of Pharmaceutical and Biomedical Analysis,2009,49(2):253-266.
[7]黄昆,王文辉,李宝才,等.超高效液相色谱和质谱联用在药物研究领域的应用[J].光谱实验室,2009,26(4):922-930.HUANG Kun,WANG Wenhui,LI Baocai,et al.Applications of combined ultra-high performance liquid chromatography(UPLC)and mass spectrometry(MS)in the field of drug research[J].Chinese Journal of Spectroscopy Laboratory,2009,26(4):922-930(in Chinese).
[8]练杭芸,徐王彦君,梁乾德,等.UPLC-QTOF MS法比较赤芍、白芍煎液化学成分差异[J].质谱学报,2014,35(3):269-278.LIAN Hangyun,XU Wangyanjun,LIANG Qiande,et al.Chemical comparison between decoctions of Radix Paeoniae Rubra and Radix Paeoniae Alba by UPLC-QTOF MS[J].Journal of Chinese Mass Spectrometry Society,2014,35(3):269-278(in Chinese).
[9]侯娅,马阳,邹立思,等.基于UPLC-Triple-TOFMS/MS技术分析不同产地太子参的差异化学成分[J].质谱学报,2015,36(4):359-366.HOU Ya,MA Yang,ZOU Lisi,et al.Difference of chemical compositions in Pseudostellariae Radix from different origins by UPLC-Triple TOF-MS/MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(4):359-366(in Chinese).
[10]王静哲,刘震,马立满,等.基于UPLC-Q-TOF MS分析加工炮制对玄参化学成分的影响[J].质谱学报,2016,37(1):1-9.WANG Jingzhe,LIU Zhen,MA Liman,et al.Changes of chemical constituents in Scrophulariae Radix during processing based on UPLC-QTOF MS[J].Journal of Chinese Mass Spectrometry Society,2016,37(1):1-9(in Chinese).
[11]张小平,蒋可志,吕惠卿,等.HPLC-Q-TOF MS鉴定条叶榕根茎乙酸乙酯提取物中的主要化学成分[J].质谱学报,2015,36(4):310-320.ZHANG Xiaoping,JIANG Kezhi,LV Huiqing,et al.Identification and characterization of major chemical compounds in the ethyl acetate extract from Ficus pandurata Hance Aerial Roots by HPLC-Q-TOF MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(4):310-320(in Chinese).
[12]宋燕燕,李敏,吴柱樟,等.采用UPLC-Q-TOF MS技术分析坛紫菜中脂质成分[J].质谱学报,2015,36(5):425-433.SONG Yanyan,LI Min,WU Zhuzhang,et al.Rapid identification of lipid compositions from Porphyra haitanensis by UPLC-Q-TOF MS[J].Journal of Chinese Mass Spectrometry Society,2015,36(5):425-433(in Chinese).
[13]马强,白桦,王超,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查化妆品中18种香豆素类化合物[J].分析测试学报,2014,33(3):248-255.MA Qiang,BAI Hua,WANG Chao,et al.Rapid screening of 18coumarin compounds in cosmetics by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J].Journal of Instrumental Analysis,2014,33(3):248-255(in Chinese).
[14]董静,王弘,万乐人,等.高效液相色谱/电喷雾-离子阱-飞行时间质谱分析鉴定中药虎杖中的主要化学成分[J].色谱,2009,27(4):425-430.DONG Jing,WANG Hong,WAN Leren,et al.Identification and determination of major constituents in Polygonum cuspidatum Sieb.et Zucc.by high performance liquid chromatography/electrospray ionization-ion trap-time-of-flight mass spectrometry[J].Chinese Journal of Chromatography,2009,27(4):425-430(in Chinese).