基于HPLC-DAD特征图谱分析莱菔子饮片酶解过程中化学成分的变化
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摘要
目的:分析莱菔子饮片酶解过程中化学成分的动态变化。方法:采用HPLC分析莱菔子不同酶解时间的特征图谱,色谱条件为流动相乙腈(A)-0.1%磷酸水溶液(B)梯度洗脱(0~15 min,5%~10%A; 15~17 min,10%~12.5%A; 17~27 min,12.5%~14%A; 27~55 min,14%~25%A; 55~57 min,25%~70%A; 57~70 min,70%~100%A),检测波长225 nm。标定其特征峰,同时提取各特征峰的UV光谱,比较不同特征峰UV光谱的异同和HPLC特征图谱特征峰峰面积的变化,分析莱菔子中特征成分的动态变化规律。结果:莱菔子不同酶解时间的特征图谱中共标定11个特征峰,归属了萝卜苷和芥子碱硫氰酸盐2个色谱峰,同时发现莱菔子中萝卜苷会快速被黑芥子酶酶解,先被酶解为一中间体,继而被分解为其他成分。芥子碱硫氰酸盐随着酶解时间没发生明显的变化。芥子酸苷类成分也会被酶解。结论:莱菔子的酶解不单是萝卜苷的酶解,还包括芥子酸苷类成分的酶解,可为莱菔子"生熟异治,生升熟降"的药性变化提供参考。
Objective:To analyze the dynamic changes of components in the enzymolysis process of raw products of Raphani Semen.Method:HPLC was employed to analysis of characteristic spectra of Raphani Semen at different enzymolysis time with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 225 nm.The characteristic peaks were calibrated,meanwhile,the UV spectra of characterstic peaks were extracted,and the difference between UV spectra and the changes of peak areas were compared,and the dynamic changes of characterstic components in Raphani Semen were analyzed.Result:Eleven characteristic peaks were marked from the characteric spectra of raw products of Raphani Semen at different enzymolysis time,and glucoraphenin and sinapine thiocyanate were assigned.Glucoraphenin was enzymatically hydrolyzed fastly by myrosinase,and an intermediate was generated,and then continue to be decomposed into other components.Sinapine thiocyanate did not change significantly during the enzymolysis process,and sinadiosides was also enzymatically degraded.Conclusion:The enzymolysis of Raphani Semen is not only the glucoraphenin,but also the sinadiosides.This paper can provide reference for the property change of Raphani Semen in processing.
Objective:To analyze the dynamic changes of components in the enzymolysis process of raw products of Raphani Semen.Method:HPLC was employed to analysis of characteristic spectra of Raphani Semen at different enzymolysis time with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 225 nm.The characteristic peaks were calibrated,meanwhile,the UV spectra of characterstic peaks were extracted,and the difference between UV spectra and the changes of peak areas were compared,and the dynamic changes of characterstic components in Raphani Semen were analyzed.Result:Eleven characteristic peaks were marked from the characteric spectra of raw products of Raphani Semen at different enzymolysis time,and glucoraphenin and sinapine thiocyanate were assigned.Glucoraphenin was enzymatically hydrolyzed fastly by myrosinase,and an intermediate was generated,and then continue to be decomposed into other components.Sinapine thiocyanate did not change significantly during the enzymolysis process,and sinadiosides was also enzymatically degraded.Conclusion:The enzymolysis of Raphani Semen is not only the glucoraphenin,but also the sinadiosides.This paper can provide reference for the property change of Raphani Semen in processing.
引文
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[13]Egner P A,CHEN J G,WANG J B,et al.Bioavailability of sulforaphane from two broccoli sprout beverages:results of a short term,cross-over clinical trial in Qidong,China[J].Cancer Prev Res(Phila),2011,4(3):384-395.
[14]Jadoun J,Yazbak A,Rushrush S,et al.Identification of a new antibacterial sulfur compound from Raphanus sativus seeds[J].Evid Based Complement Alternat Med,2016,doi:10.1155/2016/9271285.
[15]Royston K J,Tollefsbol T O.The epigenetic impact of cruciferous vegetables on cancer prevention[J].Curr Pharmacol Rep,2015,1(1):46-51.
[16]唐健元,张磊,彭成,等.莱菔子行气消食的机制研究[J].中国中西医结合消化杂志,2003,11(5):287-289.
[17]吕文海,任涛,苏永汶,等.炮制抑制莱菔子中萝卜苷酶解转化的初步实验研究[J].中国中药杂志,2011,36(8):980-983.
[18]Malik M S,Riley M B,Norsworthy J K,et al.Glucosinolate profile variation of growth stages of wild radish(Raphanus raphanistrum)[J].J Agric Food Chem,2010,58(6):3309-3315.
[19]宁永成.有机化合物结构鉴定与有机波谱学[M].北京:科学出版社,2008:369.
[2]苏慧,岳琳,刘颖,等.芥子及莱菔子饮片炮制前后物质基础变化规律分析[J].中国实验方剂学杂志,2018,24(7):23-26.
[3]ZHANG X,LIU H B,JIA J J,et al.Two novel sulfur compounds from the seeds of Raphanus sativus L.[J].JAsian Nat Prod Res,2010,12(2):113-118.
[4]蒋海强,丁臻,周洪雷.高效液相色谱测定莱菔子总碱中芥子碱硫氰酸盐含量[J].中国实验方剂学杂志,2012,18(17):78-80.
[5]孙忠迪,王群,李书云,等.炮制对莱菔子中脂肪油的含量影响及GC-MS分析[J].中国实验方剂学杂志,2013,19(1):67-69.
[6]金洪光,李玉全,凌云,等.莱菔子化学成分的分离与鉴定[J].食品科学,2018,39(6):35-40.
[7]Sham T T,Yuen A C,Ng Y F,et al.A review of the phytochemistry and pharmacological activities of Raphani Semen[J].Evid Based Complement Alternat Med,2013,doi:10.1155/2013/636194.
[8]LI Y L,YANG W Q,ZHU Q J,et al.Protective effects on vascular endothelial cell in N'-nitro-L-arginine(L-NNA)-induced hypertensive rats from the combination of effective components of Uncaria rhynchophylla and Semen Raphani[J].Biosci Trends,2015,9(4):237-244.
[9]杨金果,李运伦,周洪雷.钩藤和莱菔子生物碱抗高血压血管内皮细胞损伤效应[J].中成药,2013,35(5):889-893.
[10]张国侠,盖国忠.莱菔子总生物碱对Apo E基因敲除小鼠血脂的影响[J].中国老年学杂志,2010,30(6):844-845.
[11]ZHANG H Q,CHEN S Y,WANG A S,et al.Sulforaphane induces adipocyte browning and promotes glucose and lipid utilization[J].Mol Nutr Food Res,2016,60(10):2185-2197.
[12]谭鹏,薛玲,吕文海.莱菔子不同炮制品对呼吸系统作用的实验研究[J].山东中医杂志,2005,24(5):300-302.
[13]Egner P A,CHEN J G,WANG J B,et al.Bioavailability of sulforaphane from two broccoli sprout beverages:results of a short term,cross-over clinical trial in Qidong,China[J].Cancer Prev Res(Phila),2011,4(3):384-395.
[14]Jadoun J,Yazbak A,Rushrush S,et al.Identification of a new antibacterial sulfur compound from Raphanus sativus seeds[J].Evid Based Complement Alternat Med,2016,doi:10.1155/2016/9271285.
[15]Royston K J,Tollefsbol T O.The epigenetic impact of cruciferous vegetables on cancer prevention[J].Curr Pharmacol Rep,2015,1(1):46-51.
[16]唐健元,张磊,彭成,等.莱菔子行气消食的机制研究[J].中国中西医结合消化杂志,2003,11(5):287-289.
[17]吕文海,任涛,苏永汶,等.炮制抑制莱菔子中萝卜苷酶解转化的初步实验研究[J].中国中药杂志,2011,36(8):980-983.
[18]Malik M S,Riley M B,Norsworthy J K,et al.Glucosinolate profile variation of growth stages of wild radish(Raphanus raphanistrum)[J].J Agric Food Chem,2010,58(6):3309-3315.
[19]宁永成.有机化合物结构鉴定与有机波谱学[M].北京:科学出版社,2008:369.