整合UPLC-QTOF-MS/MS全扫描和模拟MRM方法综合评价茯苓乙醇提取物与后续乙酸乙酯萃取物三萜酸类组分化学一致性
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摘要
本研究采用超高效液相色谱串联四级杆飞行时间质谱法(UPLC-QTOF-MS/MS)对茯苓乙醇提取物与后续乙酸乙酯萃取物中三萜酸类成分的化学一致性进行评价。通过全扫描高分辨质谱数据分析,对茯苓的乙醇提取物和后续乙酸乙酯萃取物中的化学组成进行定性比较,结合文献及对照品数据对照,共鉴定出23个主要三萜酸类成分。利用模拟三重四级杆质谱(TQ-MS)的多反应监控(mMRM)模式对上述23个主要成分进行定量或相对定量分析,并计算三萜酸类组分在萃取过程中的转移率和富集率。结果表明,乙酸乙酯萃取物与乙醇提取物化学组成基本一致,萃取过程中三萜酸类成分转移率高,通过萃取可有效富集三萜酸类组分。本研究不但为科学评估后续乙酸乙酯萃取茯苓三萜酸类组分的必要性提供依据,而且为综合评价中药提取物在萃取前后的化学一致性提供了参考方法。
An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS/MS)method was developed to evaluate the chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof.First,high resolution mass spectrometry data were obtained with Full scan mode,by comparing with MS data from the reference compounds and literatures,a total of 23 components were unequivocally or tentatively identified in ethanol extracts and acetic ether extracts thereof.Then,a mimic multiple reaction monitoring(mMRM)mode was established using UPLC-QTOF-MS/MS to quantify the triter-pene acids in ethanol extracts and acetic ether extracts thereof.Eleven components were absolutely quantified with reference compounds,while 12 components without reference compounds were relatively quantified with peak areas,the transfer and enrichment rate of triterpene acids during liquid-liquid extraction were calculated.It was found all of the 23 triterpene acids identified in Poria ethanol extracts could be transferred into acetic ether extracts with high transfer and enrichment rate.The present study provides not only scientific evidence for further extrac-tion of triterpene acids in Poria by acetic ether,but also an approach for comprehensive evaluation of the chemical consistency of herbal medicine extracts before and after the liquid-liquid extraction.
An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS/MS)method was developed to evaluate the chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof.First,high resolution mass spectrometry data were obtained with Full scan mode,by comparing with MS data from the reference compounds and literatures,a total of 23 components were unequivocally or tentatively identified in ethanol extracts and acetic ether extracts thereof.Then,a mimic multiple reaction monitoring(mMRM)mode was established using UPLC-QTOF-MS/MS to quantify the triter-pene acids in ethanol extracts and acetic ether extracts thereof.Eleven components were absolutely quantified with reference compounds,while 12 components without reference compounds were relatively quantified with peak areas,the transfer and enrichment rate of triterpene acids during liquid-liquid extraction were calculated.It was found all of the 23 triterpene acids identified in Poria ethanol extracts could be transferred into acetic ether extracts with high transfer and enrichment rate.The present study provides not only scientific evidence for further extrac-tion of triterpene acids in Poria by acetic ether,but also an approach for comprehensive evaluation of the chemical consistency of herbal medicine extracts before and after the liquid-liquid extraction.
引文
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[17]Wu LF,Wang KF,Mao X,et al.Screening and analysis of the potential bioactive components of Poria cocos(Schw.)Wolf by HPLC and HPLC-MSn with the aid of chemometrics[J].Molecules,2016,21:18.
[18]Liu XT,Winkler AL,Schwan WR,et al.Antibacterial compounds from mushrooms II:lanostane triterpenoids and an ergostane steroid with activity against Bacillus cereus isolated from Fomi-topsis pinicola[J].Planta Med,2010,76:464-466.
[19]Pasandide B,Khodaiyan F,Mousavi ZE,et al.Optimization of aqueous pectin extraction from Citrus medica peel[J].Carbohydr Polym,2017,178:27-33.
[20]Zhu L,Xu J,Zhang S,et al.Qualitatively and quantitatively comparing secondary metabolites in three medicinal parts derived from Poria cocos(Schw.)Wolf using UHPLC-QTOF-MS/MS-based chemical profiling[J].J Pharm Biomed Anal,2018,150:278-286.
[2]Yu SJ,Tseng J.Fu-ling,a chinese herbal drug,modulates cytokine secretion by human peripheral blood monocytes[J].Int J Immunopharmacol,1996,18:37-44.
[3]Jeong JW,Lee HH,Han MH,et al.Ethanol extract of Poria cocos reduces the production of inflammatory mediators by suppressing the NF-κB signaling pathway in lipopolysaccharidestimulated RAW 264.7 macrophages[J].BMC Complement Altern Med,2014,14:101.
[4]Akihisa T,Nakamura Y,Tokuda H,et al.Triterpene acids from Poria cocos and their anti-tumor-promoting effects[J].J Nat Prod,2007,70:948-953.
[5]Wang DL,Chen WD,Xu XX.Anti-inflammatory effect of triter-pene acids from Poria cocos[J].Anhui Med Pharm J(安徽医药),2009,13:1021-1023.
[6]Zhang XS,Rao ZG,Hu P,et al.Preventive effect of triterpenes from Poria cocos on liver injury in mice[J].Food Sci(食品科学),2012,33:270-273.
[7]Zhang LQ,Jia Y,Luo J,et al.UPLC determination of determine dehydrotumulosic acid etc.six active compounds in Poria[J].Chin Pharm J(中国药学杂志),2012,47:1080-1083.
[8]Che S,Li Q,Huo YS,et al.RP-HPLC simultaneous determina-tion of five triterpenoid acids in different parts of Poria cocos by UV wavelengths switch[J].Acta Pharm Sin(药学学报),2010,45:494-497.
[9]Liu KY,Song YG,Liu YL,et al.An integrated strategy using UPLC-QTOF-MSE and UPLC-QTOF-MRM(enhanced target)for pharmacokinetics study of wine processed Schisandra Chinensis fructus in rats[J].J Pharm Biomed Anal,2017,139:165-178.
[10]Chindarkar NS,Park HD,Stone JA,et al.Comparison of different time of flight-mass spectrometry modes for small molecule quantitative analysis[J].J Anal Toxicol,2015,39:675-685.
[11]Zheng Y,Yang XW.Two new lanostane triterpenoids from Poria cocos[J].J Asian Nat Prod Res,2008,10:289-292.
[12]Akihisa T,Uchiyama E,Kikuchi T,et al.Anti-tumor-promoting effects of 25-methoxyporicoic acid A and other triterpene acids from Poria cocos[J].J Nat Prod,2009,72:1786-1792.
[13]Ukiya M,Akihisa T,Tokuda H,et al.Inhibition of tumorpromoting effects by poricoic acids G and H and other lanostane-type triterpenes and cytotoxic activity of poricoic acids A and G from Poria cocos[J].J Nat Prod,2002,65:462-465.
[14]Xia B,Zhou Y,Tan HS,et al.Advanced ultra-performance liquid chromatography-photodiode array-quadrupole time-of-flight mass spectrometric methods for simultaneous screening and quantifi-cation of triterpenoids in Poria cocos[J].Food Chem,2014,152:237-244.
[15]Zhao Y,Li SQ,Li HJ,et al.Lanostane triterpenoids from the fungus Ceriporia lacerate associated with Acanthaster planci[J].Chem Nat Compd,2013,49:653-656.
[16]Cai TG,Cai Y.Triterpenes from the fungus Poria cocos and their inhibitory activity on nitric oxide production in mouse macro-phages via blockade of activating protein-1 pathway[J].Chem Biodivers,2011,8:2135-2143.
[17]Wu LF,Wang KF,Mao X,et al.Screening and analysis of the potential bioactive components of Poria cocos(Schw.)Wolf by HPLC and HPLC-MSn with the aid of chemometrics[J].Molecules,2016,21:18.
[18]Liu XT,Winkler AL,Schwan WR,et al.Antibacterial compounds from mushrooms II:lanostane triterpenoids and an ergostane steroid with activity against Bacillus cereus isolated from Fomi-topsis pinicola[J].Planta Med,2010,76:464-466.
[19]Pasandide B,Khodaiyan F,Mousavi ZE,et al.Optimization of aqueous pectin extraction from Citrus medica peel[J].Carbohydr Polym,2017,178:27-33.
[20]Zhu L,Xu J,Zhang S,et al.Qualitatively and quantitatively comparing secondary metabolites in three medicinal parts derived from Poria cocos(Schw.)Wolf using UHPLC-QTOF-MS/MS-based chemical profiling[J].J Pharm Biomed Anal,2018,150:278-286.