基于非靶向代谢组学分析手工与机器煎药对八珍汤中化学成分的影响
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摘要
目的:通过非靶向代谢组学方法研究手工与机器煎煮对八珍汤中化学成分的影响,寻找这2种不同煎药方式之间的差异性化学成分。方法:以规范化手工煎药与机器煎药2种方式对八珍汤进行煎煮,利用正交偏最小二乘法-判别分析(OPLS-DA)等多元统计方法,结合变量重要性投影(VIP)值及t检验分析2种煎药方式对该复方中化学成分的影响。通过UPLC-LTQ-Orbitrap MS对差异性化合物进行分析,流动相乙腈(A)-0.1%甲酸水溶液(B)梯度洗脱(0～3 min,5%A; 3～7 min,5%～8%A; 7～9 min,8%～15%A; 9～17 min,15%～25%A; 17～27 min,25%～35%A; 27～29 min,35%～40%A; 29～35 min,40%～95%A),柱温30℃,流速0.4 mL·min-1;采用电喷雾离子源和正、负离子模式,扫描范围m/z 50～1 500。结果:在正、负离子监测模式下,共发现87个差异性成分,通过质谱数据结合文献报道,共识别并鉴定了其中的40个成分,包括洋川芎内酯A,甘草素,阿魏酸等。结论:手工与机器煎药获得的八珍汤煎煮液在色泽及化学成分上存在着较为明显的差异。若将二者的优点结合起来,在保持手工煎药优势的基础上,借鉴机器煎药包装药液的方式,建立规范的煎药流程,将使中药汤剂的药效得到最大程度上的发挥。
Objective:To evaluate the effect of manual decocting and machine decocting on the chemical constituents in Bazhentang based on non-targeted metabolomics,and to find the differential chemical constituents of these two decocting methods.Method:Bazhentang was boiled by standardized manual decocting and machine decocting methods,respectively.Orthogonal partial least square-discriminate analysis(OPLS-DA) and other multivariate statistical methods,combined with variable importance in the projection(VIP) value and t-test,were employed to analyze the effect of two decocting methods on the chemical constituents in Bazhentang.The differential chemical constituents were analyzed by UPLC-LTQ-Orbitrap MS under positive and negative ion modes,mobile phase was acetonitrile-0.1% formic acid aqueous solution for gradient elution,the scanning range was m/z 50-1 500.Result:Under the positive and negative ion modes of high-resolution mass spectrometry,a total of 87 differential components were found,40 of them were identified according to the mass spectrometry data and literature reports,including senkyunolide A,glycyrrhizin,ferulic acid,etc.Conclusion:Based on the analysis of color and chemical compositions of Bazhentang,there are obvious differences between the standardized manual decocting and machine decocting.If the advantages of these two methods are combined,a standardized decoction process can be established on the basis of maintaining the advantages of manual decocting,the effectiveness of traditional Chinese medicine decoction will be maximized and it will be convenient for patients to take it.
Objective:To evaluate the effect of manual decocting and machine decocting on the chemical constituents in Bazhentang based on non-targeted metabolomics,and to find the differential chemical constituents of these two decocting methods.Method:Bazhentang was boiled by standardized manual decocting and machine decocting methods,respectively.Orthogonal partial least square-discriminate analysis(OPLS-DA) and other multivariate statistical methods,combined with variable importance in the projection(VIP) value and t-test,were employed to analyze the effect of two decocting methods on the chemical constituents in Bazhentang.The differential chemical constituents were analyzed by UPLC-LTQ-Orbitrap MS under positive and negative ion modes,mobile phase was acetonitrile-0.1% formic acid aqueous solution for gradient elution,the scanning range was m/z 50-1 500.Result:Under the positive and negative ion modes of high-resolution mass spectrometry,a total of 87 differential components were found,40 of them were identified according to the mass spectrometry data and literature reports,including senkyunolide A,glycyrrhizin,ferulic acid,etc.Conclusion:Based on the analysis of color and chemical compositions of Bazhentang,there are obvious differences between the standardized manual decocting and machine decocting.If the advantages of these two methods are combined,a standardized decoction process can be established on the basis of maintaining the advantages of manual decocting,the effectiveness of traditional Chinese medicine decoction will be maximized and it will be convenient for patients to take it.
引文
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[13]冒海蕾,徐旻,王斌,等.正交信号校正在正常成人血清1H-NMR谱的代谢组分析中的滤噪作用评价[J].化学学报,2007,65(2):152-158.
[14] LIN L Z,HE X G,LIAN L Z,et al. Liquid chromatographic-electrospray mass spectrometric study of the phthalides of Angelica sinensis and chemical changes of Z-ligustilide[J]. J Chromatogr A,1998,810(1/2):71-79.
[15]倪书茂,钱大玮,尚尔鑫,等.大川芎方化学成分的超高效液相色谱-电喷雾-四极杆飞行时间质谱分析[J].中国实验方剂学杂志,2010,16(1):39-45.
[2]李时珍.本草纲目(上)[M].刘衡如,刘山永,校注.北京:华夏出版社,2009:297.
[3]李明森,曹文龙,王浩宇,等.中药煎煮机的现状与发展趋势[J].中国医疗器械信息,2017,23(17):29-31.
[4] Fiehn O. Metabolomics——the link between genotypes and phenotypes[J]. Plant Mol Biol,2002,48(1/2):155-171.
[5]林俊芝,张定堃,裴瑾,等.基于植物代谢组学方法区分安县附子与江油附子质量的化学标志物[J].中国实验方剂学杂志,2017,23(13):35-40.
[6]郭慧,崔扬,王秋红,等.基于代谢组学技术的中药复方研究近况[J].中国实验方剂学杂志,2017,23(1):213-219.
[7]闫利利,史家文,王金芳,等.基于UPLC/Q-TOF-MSE方法分析半夏泻心汤的化学成分[J].药学学报,2013,48(4):526-531.
[8]王镇方,赵阳,庞旭,等.利用UPLC-Q-TOF-MSE快速分析四物汤中的化学成分[J].中国中药杂志,2013,38(21):3702-3708.
[9]彭怀仁,项平.中医方剂大辞典精选本.上册[M].北京:人民卫生出版社,1999:55.
[10]王新.八珍汤临床应用体会[J].实用中医药杂志,2006,22(11):708-709.
[11]周春玉,曾庆红.八珍汤治疗气血虚弱型痛经的临床研究[J].中国中医药现代远程教育,2017,15(19):60-62.
[12]漆小泉,王玉兰,陈晓亚.植物代谢组学:方法与应用[M].北京:化学工业出版社,2011:13-18.
[13]冒海蕾,徐旻,王斌,等.正交信号校正在正常成人血清1H-NMR谱的代谢组分析中的滤噪作用评价[J].化学学报,2007,65(2):152-158.
[14] LIN L Z,HE X G,LIAN L Z,et al. Liquid chromatographic-electrospray mass spectrometric study of the phthalides of Angelica sinensis and chemical changes of Z-ligustilide[J]. J Chromatogr A,1998,810(1/2):71-79.
[15]倪书茂,钱大玮,尚尔鑫,等.大川芎方化学成分的超高效液相色谱-电喷雾-四极杆飞行时间质谱分析[J].中国实验方剂学杂志,2010,16(1):39-45.