固相萃取结合UHPLC-LTQ-Orbitrap MS分析黄精发酵前后的化学成分
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摘要
目的应用固相萃取结合超高效液相色谱-线性离子阱-静电场轨道质谱(UHPLC-LTQ-Orbitrap MS)技术对黄精发酵前后的化学成分进行分析鉴定。方法采用Waters HSS T3(100 mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水溶液和的乙腈为流动相梯度洗脱,体积流量0.3 mL/min,在电喷雾正离子模式下进行检测,四级杆飞行时间串联质谱法对各主要色谱峰进行归属。结果根据所获得的精确相对分子质量,同时结合色谱保留行为、质谱裂解规律、特征碎片离子、对照品比对以及相关文献报道,共鉴定发酵前后黄精中的69个化学成分,包括57个甾体皂苷类成分、5个黄酮类成分、4个三萜皂苷类成分、2个生物碱类及1个有机酸类成分。其中,从发酵前黄精提取液中鉴定了62个化学成分,从发酵后黄精提取液中鉴定了18个化学成分。结论发酵对黄精中的化学成分影响很大,如成分异构化、原生苷种类以及含量减少、次生苷或苷元含量的增多等,可为黄精的进一步开发利用提供科学依据。
Objective To analyze and identify the chemical constituents in aqueous extracts and fermentation broth from Polygonatum sibiricum by ultra performance liquid chromatography-linear ion trap-electrostatic field orbital mass spectrometry(UHPLC-LTQOrbitrap MS) combined with solid phase extraction. Methods The separation was performed on Waters HSS T3(100 mm × 2.1 mm,1.7 μm) with acetonitrile and 0.1% formic acid solution as the mobile phase in gradient elution. The flow rate was 0.3 mL/min. The detection was carried out in electrospray positive ion mode, and the main peaks were assigned by quadrupole time-of-flight mass spectrometry. Results According to the obtained accurate molecular weight, combined with chromatographic retention behavior,mass spectrometry cleavage, characteristic fragment ions, reference comparison, and related literature reports, a total of 69 chemical components were identified in aqueous extracts and fermentation broth from P. sibiricum, including 57 steroidal saponins, five flavonoid components, four triterpenoid saponins, two alkaloids, and one organic acid component. Among them, 62 chemical components were identified from the aqueous extracts, and 18 chemical components were identified from the fermentation broth of P.sibiricum. Conclusion The results showed that fermentation has a great influence on the chemical components of P. sibiricum,including the isomerization of components, types and content reduction of primary glycosides and the concentration of components changed of secondary glycosides in different degrees, which provides the necessary scientific basis for the further development of P. sibiricum.
Objective To analyze and identify the chemical constituents in aqueous extracts and fermentation broth from Polygonatum sibiricum by ultra performance liquid chromatography-linear ion trap-electrostatic field orbital mass spectrometry(UHPLC-LTQOrbitrap MS) combined with solid phase extraction. Methods The separation was performed on Waters HSS T3(100 mm × 2.1 mm,1.7 μm) with acetonitrile and 0.1% formic acid solution as the mobile phase in gradient elution. The flow rate was 0.3 mL/min. The detection was carried out in electrospray positive ion mode, and the main peaks were assigned by quadrupole time-of-flight mass spectrometry. Results According to the obtained accurate molecular weight, combined with chromatographic retention behavior,mass spectrometry cleavage, characteristic fragment ions, reference comparison, and related literature reports, a total of 69 chemical components were identified in aqueous extracts and fermentation broth from P. sibiricum, including 57 steroidal saponins, five flavonoid components, four triterpenoid saponins, two alkaloids, and one organic acid component. Among them, 62 chemical components were identified from the aqueous extracts, and 18 chemical components were identified from the fermentation broth of P.sibiricum. Conclusion The results showed that fermentation has a great influence on the chemical components of P. sibiricum,including the isomerization of components, types and content reduction of primary glycosides and the concentration of components changed of secondary glycosides in different degrees, which provides the necessary scientific basis for the further development of P. sibiricum.
引文
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[23]陈辉,冯珊珊,孙彦君,等.3种药用黄精的化学成分及药理活性研究进展[J].中草药,2015,46(15):2329-2338.
[24]王易芬,穆天慧,陈纪军,等.滇黄精化学成分研究[J].中国中药杂志,2003,28(6):524-527.
[2]陈哗,孙晓生.黄精的药理研究进展[J].中药新药与临床药理,2010,21(3):328-330.
[3]张洁,马百平,杨云,等.黄精属植物甾体皂苷类成分及药理活性研究进展[J].中国药学杂志,2006,41(5):330-332.
[4]刘亮镜,潘扬.中药的发酵炮制初探[J].现代中药研究与实践,2009,23(1):2.
[5]杨婧娟,张希,谭书宇,等.黄精发酵工艺的初步研究[J].食品研究与开发,2016,37(17):81-88.
[6]黄莺.黄精皂苷对CUMS抑郁模型大鼠行为学、血清中微量元素、5-羟色胺的影响[D].合肥:安徽医科大学,2012.
[7]Cai J,Liu M,Wang Z,et al.Apoptosis induced by dioscin in Hela cells[J].Biol Pharm Bull,2002,25(2):193-196.
[8]杨崇仁,张影,王东,等.黄精属植物甾体皂苷的分子进化及其化学分类学意义[J].云南植物研究,2007,29(5):591-600.
[9]匡海学.中药化学[M].北京:中国中医药出版社,2003.
[10]丛浦珠.天然有机质谱学[M].北京:中国医药科技出版社,2003.
[11]范书珍,陈存武,王林.多花黄精总皂苷的提取研究[J].皖西学院学报,2005,21(5):39-41.
[12]Hu C Y,Xu D P,Wu Y M,et al.Triterpenoid saponins from the rhizome of Polygonatum sibiricum[J].J Asian Nat Prod Res,2010,12(9):801-808.
[13]Yu H S,Ma B P,Kang L P,et al.Saponins from the processed rhizomes of Polygonatum kingianum[J].Chem Pharm Bull,2009,57(9):1011-1014.
[14]徐德平,孙婧,齐斌,等.黄精中三萜皂苷的提取分离与结构鉴定[J].中草药,2006,37(10):1470-1472.
[15]孙隆儒,李铣.黄精化学成分的研究(II)[J].中草药,2001,32(7):586-588.
[16]张永,严安定,高建.液质联用技术鉴定甘草提取物中的主要化学成分[J].中成药,2012,34(6):1111-1116.
[17]赵艳敏,刘素香,张晨曦,等.基于HPLC-Q-TOF-MS技术的甘草化学成分分析[J].中草药,2016,47(12):2061-2068.
[18]李自红,魏悦,范毅,等.芦丁的电喷雾离子阱质谱分析[J].分析试验室,2015,(2):186-189.
[19]高颖,戚楚露,张磊,等.黄精新鲜药材的化学成分[J].药学与临床研究,2015,23(4):365-367.
[20]Son K H,Do J C,Kang S S.Isolation of adenosine from the rhizomes of Polygonatum sibiricum[J].Arch Pharm Res,1991,14(2):193-194.
[21]Sun L R,Li X,Wang S X.Two new alkaloids from the rhizome of Polygonatum sibiricum[J].J Asian Nat Prod Res,2005,7(2):127-130.
[22]Wang Y F,Lu C H,Lai G F,et al.A new indolizinone from Polygonatum kingianum[J].Planta Med,2003,69(11):1066-1068.
[23]陈辉,冯珊珊,孙彦君,等.3种药用黄精的化学成分及药理活性研究进展[J].中草药,2015,46(15):2329-2338.
[24]王易芬,穆天慧,陈纪军,等.滇黄精化学成分研究[J].中国中药杂志,2003,28(6):524-527.