基于UPLC-Q-TOF-MS法分析百合珠芽化学成分及其薯蓣皂苷元抗肿瘤活性研究
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摘要
为建立超高效液相色谱-四级杆飞行时间质谱(UPLC-Q-TOF-MS)快速鉴定百合珠芽中10种化学成分的分析方法,并对薯蓣皂苷元进行抗肿瘤研究。实验采用Thermo Hypersil Gold C_(18)柱(100 mm×2.1 mm,1.9μm);乙腈(A)-0.5%乙酸(B)为流动相,梯度洗脱;电喷雾离子源(ESI),正负离子同步监测。采用CCK8法进行薯蓣皂苷元抗肺肿瘤A549细胞、胃肿瘤HGC-27细胞的药效筛选。结果表明,通过二级高分辨质谱分析结合相关文献,共鉴定对香豆酸、薯蓣皂苷、槲皮素等10个化合物。药理研究显示,薯蓣皂苷元对肺癌A549细胞增殖只有微弱的抑制活性,而对胃癌HGC-27细胞增殖表现出较强的抑制作用。本试验证实UPLC-Q-TOF-MS/MS可从保留时间、紫外吸收光谱、精确相对分子质量、分子式和二级结构碎片等方面对百合珠芽的主要成分进行定性分析,为寻找百合珠芽中活性物质提供一种快速准确的分析方法,且揭示百合珠芽中鉴定得到的单体成分可能具有较强的抗肿瘤活性。
To establish a method for rapid identification of 10 components in bulbil of Lilium lancifolium Thunb.by UPLC-Q-TOF-MS,and carry out research of tumor cell proliferation study of diosgenin.A Thermo Hypersil Gold C_(18) column(100 mm×2.1 mm,1.9 μm) was used with the mobile phase of acetonitrile(A)-0.5% acetic acid(B) by gradient elution.Positive and negative ions were simultaneously monitored by electro-spray ionization(ESI),and CCK-8 assay was used to screen the effect of diosgenin on lung cancer A549 cells and gastric cancer HGC-27 cells.Based on the fragmentation pattern and reference results,the structures of 10 compounds including p-coumaric acid,diosgenin and quercetin were identified.The pharmacological study showed that diosgenin had only a weak inhibitory activity on the proliferation of A549 cells in lung cancer,and had a strong inhibitory effect on the proliferation of HGC-27 cells in gastric cancer.These results suggested that UPLC-Q-TOF-MS/MS can be used in the qualitative analysis of the main components of bulbil of Lilium lancifolium Thunb.from retention time,UV absorption spectrum,precise relative molecular weight,molecular formula and secondary fragment ions,and so on.Meanwhile,diosgenin have a strong inhibitory effect on the proliferation of gastric cancer HGC-27 cells.This research provided a scientific basis for the ues of bulbil of Lilium lancifolium Thunb~1..
To establish a method for rapid identification of 10 components in bulbil of Lilium lancifolium Thunb.by UPLC-Q-TOF-MS,and carry out research of tumor cell proliferation study of diosgenin.A Thermo Hypersil Gold C_(18) column(100 mm×2.1 mm,1.9 μm) was used with the mobile phase of acetonitrile(A)-0.5% acetic acid(B) by gradient elution.Positive and negative ions were simultaneously monitored by electro-spray ionization(ESI),and CCK-8 assay was used to screen the effect of diosgenin on lung cancer A549 cells and gastric cancer HGC-27 cells.Based on the fragmentation pattern and reference results,the structures of 10 compounds including p-coumaric acid,diosgenin and quercetin were identified.The pharmacological study showed that diosgenin had only a weak inhibitory activity on the proliferation of A549 cells in lung cancer,and had a strong inhibitory effect on the proliferation of HGC-27 cells in gastric cancer.These results suggested that UPLC-Q-TOF-MS/MS can be used in the qualitative analysis of the main components of bulbil of Lilium lancifolium Thunb.from retention time,UV absorption spectrum,precise relative molecular weight,molecular formula and secondary fragment ions,and so on.Meanwhile,diosgenin have a strong inhibitory effect on the proliferation of gastric cancer HGC-27 cells.This research provided a scientific basis for the ues of bulbil of Lilium lancifolium Thunb~1..
引文
1 Chinese Pharmacopoeia Commission.Pharmacopoeia of the People’s Republic of China:Vol Ⅰ(中华人民共和国药典:第一部)[M].Beijing:China Medical Science Press,2015:132.
2 Fu CY,Liu YH,Li MJ,et al.Extraction and purification techniques and antidepression activity of total saponins from bulbs of Lilium brownii[J].Nat Prod Res Dev(天然产物研究与开发),2012,24:682-686.
3 Zhu Y.Extraction,purification,identification and antioxidant activity of the active ingredients from the bulbil and bulb of Lilium brownii var.viridulum by LC-ESI-MS/MS[D].Nanjing:Jiangsu University(江苏大学),2016.
4 Yuan ZY,Liu XD,Pei G,et al.Contents determination of p-coumaric acid and gallic acid in lilii bulbus by HPLC[J].Chin J Inf Tradit Chin Med(中国中医药信息杂志),2018,25(5):82-85.
5 Guo QP,Gao Y,Li WM.HPLC fingerprint of Lilium brownii[J].Chin Tradit Pat Med(中成药),2011,33:1280-1285.
6 Zhang ZJ,Cai BC Li L,et al.GC/MS fingerprint of Bulbus lilii.[J].Chin Tradit Pat Med(中成药),2006,28:625-627.
7 Wang TT.Studies on chemical constituents of Lilium and Pyrus cultivars[D].Tianjin:Tianjin University(天津大学)2015.
8 Zhang M,Duan C,Zang Y,et al.The flavonoid composition of flavedo and juice from the pummelo cultivar Citrus grandis,(L.) Osbeck and the grapefruit cultivar Citrus paradisi from China[J].Food Chem,2011,129:1530-1536.
9 Xiang Q,Wang XH,Lin H,et al.Qualitative and quantitative analysis of major constituents in Yinhuang granules by HPLC-DAD-Q-TOF-MS / MS[J].Chin Tradit Pat Med(中成药),2015,37:105-112.
10 Zhang HQ,Yan H,Qian DW,et al.Analysis and evaluation of eight active ingredients in lilium lancifolium from different regions[J].Chin J Chin Mater Med(中国中药杂志),2017,42:311-318.
11 Nie H,Yan H,Qian DW,et al.Study on UPLC specific chromatogram of lily and its specific peaks compositions analysis by QTOF-MS[J].Chin Med Mat(中药材),2013,36:1087-1092.
12 Zhang Y,Feng BM,Lu X.Research progress on application of UPLC/Q-TOF-MS in pharmaceutical analysis[J].Nat Prod Res Dev(天然产物研究与开发),2017,29:1992-1996.
13 He LL,Wang X,Gu KB,et al.Research progress of tumor and depression disorder[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2012,8:144-146.
14 Wei YL.Studying the anti-tumor effects and the possible mechanisms of dioscin on three kinds of human lung cancer cell lines[D].Dalian:Dalian Medical University(大连医科大学)2013.
15 Wang M.The study On the inhibition of curcumol and dioscin in Qizhu decoction to the proliferation of human gastric cancer cell MGC-803[D].Nanjing:Nanjing University of Chinese Medicine,(南京中医药大学),2012.
16 Hu MM.Dioscin induces the apoptosis of human gastric carcinoma SGC-7901 cells and mechanism investigation[D].Dalian:Dalian Medical University(大连医科大学),2011.
17 Qin KM,Fang QB,Cai H,et al.Analysis of chemical components in Baihe Zhimu Tang and its single herbs by high performance liquid chromatography-electrospray ionization-mass spectrometry[J].Chin J Anal Chem(分析化学),2009,37:1759-1764.
18 Díazgarcía MC,Obón JM,Castellar MR,et al.Quantification by UHPLC of total individual polyphenols in fruit juices[J].Food Chem,2013,123:138-152.
19 Li YH,Dai HX,Wang MM,et al.Elucidation of quercetin by electron spray ionization mass spectrometry[J].J Chin Mass Spectro Soci(质谱学报),2009,30:374-378.
20 Dai XW,Feng LL,Li HF.Analysis of differences and correlation of steroidal saponins in rhizomes and leaves of paris polyphylla var.yunnanensis from different planting base[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2018,3:41-48.
2 Fu CY,Liu YH,Li MJ,et al.Extraction and purification techniques and antidepression activity of total saponins from bulbs of Lilium brownii[J].Nat Prod Res Dev(天然产物研究与开发),2012,24:682-686.
3 Zhu Y.Extraction,purification,identification and antioxidant activity of the active ingredients from the bulbil and bulb of Lilium brownii var.viridulum by LC-ESI-MS/MS[D].Nanjing:Jiangsu University(江苏大学),2016.
4 Yuan ZY,Liu XD,Pei G,et al.Contents determination of p-coumaric acid and gallic acid in lilii bulbus by HPLC[J].Chin J Inf Tradit Chin Med(中国中医药信息杂志),2018,25(5):82-85.
5 Guo QP,Gao Y,Li WM.HPLC fingerprint of Lilium brownii[J].Chin Tradit Pat Med(中成药),2011,33:1280-1285.
6 Zhang ZJ,Cai BC Li L,et al.GC/MS fingerprint of Bulbus lilii.[J].Chin Tradit Pat Med(中成药),2006,28:625-627.
7 Wang TT.Studies on chemical constituents of Lilium and Pyrus cultivars[D].Tianjin:Tianjin University(天津大学)2015.
8 Zhang M,Duan C,Zang Y,et al.The flavonoid composition of flavedo and juice from the pummelo cultivar Citrus grandis,(L.) Osbeck and the grapefruit cultivar Citrus paradisi from China[J].Food Chem,2011,129:1530-1536.
9 Xiang Q,Wang XH,Lin H,et al.Qualitative and quantitative analysis of major constituents in Yinhuang granules by HPLC-DAD-Q-TOF-MS / MS[J].Chin Tradit Pat Med(中成药),2015,37:105-112.
10 Zhang HQ,Yan H,Qian DW,et al.Analysis and evaluation of eight active ingredients in lilium lancifolium from different regions[J].Chin J Chin Mater Med(中国中药杂志),2017,42:311-318.
11 Nie H,Yan H,Qian DW,et al.Study on UPLC specific chromatogram of lily and its specific peaks compositions analysis by QTOF-MS[J].Chin Med Mat(中药材),2013,36:1087-1092.
12 Zhang Y,Feng BM,Lu X.Research progress on application of UPLC/Q-TOF-MS in pharmaceutical analysis[J].Nat Prod Res Dev(天然产物研究与开发),2017,29:1992-1996.
13 He LL,Wang X,Gu KB,et al.Research progress of tumor and depression disorder[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2012,8:144-146.
14 Wei YL.Studying the anti-tumor effects and the possible mechanisms of dioscin on three kinds of human lung cancer cell lines[D].Dalian:Dalian Medical University(大连医科大学)2013.
15 Wang M.The study On the inhibition of curcumol and dioscin in Qizhu decoction to the proliferation of human gastric cancer cell MGC-803[D].Nanjing:Nanjing University of Chinese Medicine,(南京中医药大学),2012.
16 Hu MM.Dioscin induces the apoptosis of human gastric carcinoma SGC-7901 cells and mechanism investigation[D].Dalian:Dalian Medical University(大连医科大学),2011.
17 Qin KM,Fang QB,Cai H,et al.Analysis of chemical components in Baihe Zhimu Tang and its single herbs by high performance liquid chromatography-electrospray ionization-mass spectrometry[J].Chin J Anal Chem(分析化学),2009,37:1759-1764.
18 Díazgarcía MC,Obón JM,Castellar MR,et al.Quantification by UHPLC of total individual polyphenols in fruit juices[J].Food Chem,2013,123:138-152.
19 Li YH,Dai HX,Wang MM,et al.Elucidation of quercetin by electron spray ionization mass spectrometry[J].J Chin Mass Spectro Soci(质谱学报),2009,30:374-378.
20 Dai XW,Feng LL,Li HF.Analysis of differences and correlation of steroidal saponins in rhizomes and leaves of paris polyphylla var.yunnanensis from different planting base[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2018,3:41-48.