干燥温度和提取方法对蛋白桑 1-Deoxynojirimycin提取量的影响
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摘要
为研究不同干燥温度和提取方法对1-脱氧野尻霉素(1-Deoxynojirimycin,DNJ)提取量的影响,收割新鲜的蛋白桑(Morus alba)在不同温度下干燥后,分别用酸提法、水提法、醇提法提取其中的DNJ。提取液用柱前衍生高效液相色谱荧光检测法(HPLC-FLD)测定DNJ含量。测定结果表明,采用盐酸提取法时,蛋白桑最佳干燥温度为90℃,DNJ提取量为3.38mg·g~(-1),显著高于其他温度干燥下的提取量(P<0.05);采用水提取法时,蛋白桑最佳干燥温度为80℃,DNJ提取量为1.91mg·g~(-1),显著高于其他温度干燥下的提取量(P<0.05);采用乙醇提取法时,蛋白桑最佳干燥温度为100℃,DNJ提取量为3.77mg·g~(-1),显著高于其他温度干燥下的提取量(P<0.05);对相同处理的样品,不同提取方法的提取量差异显著(P<0.05),乙醇提取法的DNJ提取量最高,盐酸提取法的DNJ提取量较高,水提取法的DNJ提取量最低。
The aim of this study was to determine the effects of different drying temperatures and extraction methods on the amount of 1-deoxynojirimycin(DNJ)extracted from Morus alba.Fresh aboveground of M.alba were harvested and dried at different temperatures.The DNJ in the samples was extracted by acid extraction,water extraction,and ethanol extraction.The DNJ content were determined by apre-column derivatization high-performance liquid chromatography system equipped with a fluorescence detector(HPLC-FLD).The results showed that when using the hydrochloric acid extraction method,the optimal drying temperature was 90℃ and the amount of DNJ extracted was 3.38 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05);when using the water extraction method,the optimum drying temperature was 80 ℃ and the amount of DNJ extracted was 1.91 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05);when using the ethanol extraction method,the optimum drying temperature was 100℃and the amount of DNJ extracted was 3.77 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05).For the samples under the same temperature treatment,the amount of DNJ extracted varied significantly across the different extraction methods(P<0.05);the order of the amount of DNJ extracted by the three extraction methods was ethanol extraction method > hydrochloric acid extraction method > water extraction method.
The aim of this study was to determine the effects of different drying temperatures and extraction methods on the amount of 1-deoxynojirimycin(DNJ)extracted from Morus alba.Fresh aboveground of M.alba were harvested and dried at different temperatures.The DNJ in the samples was extracted by acid extraction,water extraction,and ethanol extraction.The DNJ content were determined by apre-column derivatization high-performance liquid chromatography system equipped with a fluorescence detector(HPLC-FLD).The results showed that when using the hydrochloric acid extraction method,the optimal drying temperature was 90℃ and the amount of DNJ extracted was 3.38 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05);when using the water extraction method,the optimum drying temperature was 80 ℃ and the amount of DNJ extracted was 1.91 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05);when using the ethanol extraction method,the optimum drying temperature was 100℃and the amount of DNJ extracted was 3.77 mg·g~(-1),which was significantly higher than that in the other groups(P<0.05).For the samples under the same temperature treatment,the amount of DNJ extracted varied significantly across the different extraction methods(P<0.05);the order of the amount of DNJ extracted by the three extraction methods was ethanol extraction method > hydrochloric acid extraction method > water extraction method.
引文
[1]熊意,王红红,张芳松,穆建强,祝建波.蛋白桑叶片再生体系的建立及对草丁膦敏感性测定.北方园艺,2016(17):98-102.XIONG Y,WANG H H,ZHANG F S,MU J Q,ZHU J B.Establishment of protein mulberry leaves’regeneration system and sensitivity test of phosphinothricin.Northern Horticulture,2016(17):98-102.
[2]唐静,柴尧,彭九嫚,陈丽叶,杜俊潮,薛珍珍,程建明.柱前衍生化HPLC-UV法检测桑叶中1-脱氧野尻霉素含量.时珍国医国药,2016,27(5):1096-1098.TANG J,CHAI Y,PENG J M,CHEN L Y,DU J C,XUE Z Z,CHENG J M.Determination of 1-deoxynojirimycin in mulberry leaf using HPLC-UV with precolumn derivatization.Lishizhen Medicine and Materia Medica Research,2016,27(5):1096-1098.
[3]郑雪,夏旭.桑叶化学成分及其药用保健功能的研究进展.医学综述,2013,19(12):2210-2211.ZHENG X,XIA X.Research advances in chemical composition,medicinal and health care function of mulberry leaves.Medical Recapitulate,2013,19(12):2210-2211.
[4]KIMURA M,CHEN F.Antihyperglycemic effects of N-containing sugars derived from mulberry leaves in streptozoan-induced diabetic mice.Wakan Iyakugaku Zasshi,1995,12(3):214-219.
[5]杨雨,欧阳臻,常钰,孟夏.桑叶不同组分降血糖作用研究.食品科学,2007,28(8):454-456.YANG Y,OUYANG Z,CHANG Y,MENG X.Study on hypoglycemic effects of components in mulberry leaves.Food Science,2007,28(8):454-456.
[6]周晓玲,孙凌云,张进,殷浩,崔为正.1-脱氧野尻霉素的来源及合成研究进展.蚕业科学,2011,37(1):105-111.ZHOU X L,SUN L Y,ZHANG J,YIN H,CUI W Z.Research progress in origin and synthesis of 1-deoxynojirimycin.Science of Sericulture,2011,37(1):105-111.
[7]FUKAYA N,MOCHIZUKI K,TANAKA Y,KUMAZAWA T,ZHU J X.Theα-glucosidase inhibitor miglitol delays the development of diabetes and dysfunctional insulin secretion in pancreaticβ-cells in OLETF rats.European Journal of Pharmacology,2009,624(1):51-57.
[8]ZHANG Y,REN C J,LU G B,CUI W Z,MU Z M,GAO H J,WANG Y W.Purification,characterization and anti-diabeticactivity of a polysaccharide from mulberry leaf.Regulatory Toxicology and Pharmacology,2014,70(3):687-695.
[9]沈小月,宋敏,杭太俊.LC-MS法鉴定桑叶中生物碱类成分.药物分析杂志,2016,36(10):1737-1744.SHEN X Y,SONG M,HANG T J.Identification and characterization of alkaloids in mulberry leaf by hyphenated LC-MS techniques.Chinese Journal of Pharmaceutical Analysis,2016,36(10):1737-1744.
[10]彭忠田,申璀,谭德明,周应军,侯周华,朱平安.脱氧野尻霉素衍生物抗乙型肝炎病毒的体外试验研究.中国药房,2007,18(1):22-24.PENG Z T,SHEN C,TAN D M,ZHOU Y J,HOU Z H,ZHU P A.Anti-HBV activity of deoxynojirimycin derivatives in vitro.China Pharmacy,2007,18(1):22-24.
[11]LIM H H,YANG S J,KIM Y,LEE M,LIM Y.Combined treatment of mulberry leaf and fruit extract ameliorates obesity-related inflammation and oxidative stress in high fat diet-induced obese mice.Journal of Medicinal Food,2013,16(8):673-680.
[12]YANG N C,JHOU K Y,TSENG C Y.Antihypertensive effect of mulberry leaf aqueous extract containingγ-aminobutyric acid in spontaneously hypertensive rats.Food Chemistry,2012,132(4):1796-1801.
[13]KIM J Y,CHOI B G,JUNG M J,WEE J H,CHUNG K H,KWON O.Mulberry leaf water extract ameliorates insulin sensitivity in high fat or high sucrose diet induced overweight rats.Journal of the Korean Society for Applied Biological Chemistry,2011,54(4):612-618.
[14]欧阳臻,李永辉,徐卫东,陈钧.高效液相色谱-荧光检测法测定桑叶中1-脱氧野尻霉素(DNJ)含量.中国中药杂志,2005,30(9):682-685.OUYANG Z,LI Y H,XU W D,CHEN J.Determination of 1-deoxynojirimycin in leaves of Morus alba by high performance liquid chromatography with fluorescence detection.China Journal of Chinese Materia Medica,2005,30(9):682-685.
[15]欧阳臻,陈钧.不同季节桑叶中1-脱氧野尻霉素(DNJ)含量的测定.食品科学,2004,25(10):211-214.OUYANG Z,CHEN J.Determination of 1-deoxynojirimycin in Morus alba L.leaves in different seasons.Food Science,2004,25(10):211-214.
[16]曾锐,祝勇军,刘超.HPLC-ELSD法测定桑树不同部位中的1-脱氧野尻霉素.华西药学杂志,2009,24(2):171-172.ZENG R,ZHU Y J,LIU C.Determination of 1-deoxynojinimycin in different parts of mulberry by HPLC-ELSD.West China Journal of Pharmaceutical Sciences,2009,24(2):171-172.
[17]姜永涛,陈继永,刘珂.HPLC-ELSD测定桑叶中1-脱氧野尻霉素的含量.药物分析杂志,2005,25(1):27-29.JIANG Y T,CHEN J Y,LIU K.HPLC-ELSD determination of 1-deoxynojinimycin in mulberry leaves.Chinese Journal of Pharmaceutical Analysis,2005,25(1):27-29.
[18]肖洪,黄先智,沈以红,丁晓雯,张亚琼,梁菡峪,张迪.不同发酵条件对发酵桑叶茶生物活性成分含量的影响.食品科学,2013,34(23):216-220.XIAO H,HUANG X Z,SHEN Y H,DING X W,ZHANG Y Q,LIANG H Y,ZHANG D.Effects of different fermentation condition on the contents of bioactive ingredients in fermented mulberry leaf tea.Food Science,2013,34(23):216-220.
[19]胡瑞君,车振明.微波萃取桑叶1-脱氧野尻霉素工艺的研究.食品工程,2008(1):50-53.HU R J,CHE Z M.Studies on the microwave-assisted extraction method of DNJ from mulberry leaves.Food Engineering,2008(1):50-53.
[20]郑晓静,伍金娥,何计国,高月.响应面法优化超声波提取桑叶中1-脱氧野尻霉素的工艺研究.中国食品学报,2012,12(9):34-40.ZHENG X J,WU J E,HE J G,GAO Y.Optimization of 1-deoxynojirimycin extraction from mulberry leaves by using response surface methodology.Journal of Chinese Institute of Food Science and Technology,2012,12(9):34-40.
[21]刘凡,李平平,廖森泰,肖更生,邹宇晓,施英,吴娱明,唐翠明.98份不同桑树品种资源的桑叶总生物碱及1-脱氧野尻霉素含量测定.蚕业科学,2012,38(2):0185-0191.LIU F,LI P P,LIAO S T,XIAO G S,ZOU Y X,SHI Y,WU Y M,TANG C M.Determination of total alkaloids and 1-deoxynojirimycin contents in leaves of 98mulberry variety resources.Science of Sericulture,2012,38(2):0185-0191.
[22]刘伟鋆,赵骏,高秀梅,刘美平,曾森.1-脱氧野尻霉素提取精制工艺的研究.中华中医药学刊,2009,27(10):2079-2081.LIU W J,ZHAO J,GAO X M,LIU M P,ZENG S.Study on the extraction and purification of DNJ from Morus alba L.Chinese Archives of Traditional Chinese Medicine.2009,27(10):2079-2081.
[23]秦樱瑞,曾艺涛,杨娟,丁晓雯,黄先智.干燥方式对桑叶降糖活性成分含量的影响.食品科学,2015,36(17):71-76.QIN Y R,ZENG Y T,YANG J,DING X W,HUANG X Z.Effects of drying methods on the contents of hypoglycemic components in mulberry leaves(Morus alba L.).Food Science,2015,36(17):71-76.
[24]刘红英,何军,何珺.不同干燥方法对桑白皮中DNJ含量的影响.中药材,2012,35(7):1053-1055.LIU H Y,HE J,HE J.Effect of different drying methods on DNJ content in Cortex Mori.Journal of Chinese Medicinal Materials,2012,35(7):1053-1055.
[2]唐静,柴尧,彭九嫚,陈丽叶,杜俊潮,薛珍珍,程建明.柱前衍生化HPLC-UV法检测桑叶中1-脱氧野尻霉素含量.时珍国医国药,2016,27(5):1096-1098.TANG J,CHAI Y,PENG J M,CHEN L Y,DU J C,XUE Z Z,CHENG J M.Determination of 1-deoxynojirimycin in mulberry leaf using HPLC-UV with precolumn derivatization.Lishizhen Medicine and Materia Medica Research,2016,27(5):1096-1098.
[3]郑雪,夏旭.桑叶化学成分及其药用保健功能的研究进展.医学综述,2013,19(12):2210-2211.ZHENG X,XIA X.Research advances in chemical composition,medicinal and health care function of mulberry leaves.Medical Recapitulate,2013,19(12):2210-2211.
[4]KIMURA M,CHEN F.Antihyperglycemic effects of N-containing sugars derived from mulberry leaves in streptozoan-induced diabetic mice.Wakan Iyakugaku Zasshi,1995,12(3):214-219.
[5]杨雨,欧阳臻,常钰,孟夏.桑叶不同组分降血糖作用研究.食品科学,2007,28(8):454-456.YANG Y,OUYANG Z,CHANG Y,MENG X.Study on hypoglycemic effects of components in mulberry leaves.Food Science,2007,28(8):454-456.
[6]周晓玲,孙凌云,张进,殷浩,崔为正.1-脱氧野尻霉素的来源及合成研究进展.蚕业科学,2011,37(1):105-111.ZHOU X L,SUN L Y,ZHANG J,YIN H,CUI W Z.Research progress in origin and synthesis of 1-deoxynojirimycin.Science of Sericulture,2011,37(1):105-111.
[7]FUKAYA N,MOCHIZUKI K,TANAKA Y,KUMAZAWA T,ZHU J X.Theα-glucosidase inhibitor miglitol delays the development of diabetes and dysfunctional insulin secretion in pancreaticβ-cells in OLETF rats.European Journal of Pharmacology,2009,624(1):51-57.
[8]ZHANG Y,REN C J,LU G B,CUI W Z,MU Z M,GAO H J,WANG Y W.Purification,characterization and anti-diabeticactivity of a polysaccharide from mulberry leaf.Regulatory Toxicology and Pharmacology,2014,70(3):687-695.
[9]沈小月,宋敏,杭太俊.LC-MS法鉴定桑叶中生物碱类成分.药物分析杂志,2016,36(10):1737-1744.SHEN X Y,SONG M,HANG T J.Identification and characterization of alkaloids in mulberry leaf by hyphenated LC-MS techniques.Chinese Journal of Pharmaceutical Analysis,2016,36(10):1737-1744.
[10]彭忠田,申璀,谭德明,周应军,侯周华,朱平安.脱氧野尻霉素衍生物抗乙型肝炎病毒的体外试验研究.中国药房,2007,18(1):22-24.PENG Z T,SHEN C,TAN D M,ZHOU Y J,HOU Z H,ZHU P A.Anti-HBV activity of deoxynojirimycin derivatives in vitro.China Pharmacy,2007,18(1):22-24.
[11]LIM H H,YANG S J,KIM Y,LEE M,LIM Y.Combined treatment of mulberry leaf and fruit extract ameliorates obesity-related inflammation and oxidative stress in high fat diet-induced obese mice.Journal of Medicinal Food,2013,16(8):673-680.
[12]YANG N C,JHOU K Y,TSENG C Y.Antihypertensive effect of mulberry leaf aqueous extract containingγ-aminobutyric acid in spontaneously hypertensive rats.Food Chemistry,2012,132(4):1796-1801.
[13]KIM J Y,CHOI B G,JUNG M J,WEE J H,CHUNG K H,KWON O.Mulberry leaf water extract ameliorates insulin sensitivity in high fat or high sucrose diet induced overweight rats.Journal of the Korean Society for Applied Biological Chemistry,2011,54(4):612-618.
[14]欧阳臻,李永辉,徐卫东,陈钧.高效液相色谱-荧光检测法测定桑叶中1-脱氧野尻霉素(DNJ)含量.中国中药杂志,2005,30(9):682-685.OUYANG Z,LI Y H,XU W D,CHEN J.Determination of 1-deoxynojirimycin in leaves of Morus alba by high performance liquid chromatography with fluorescence detection.China Journal of Chinese Materia Medica,2005,30(9):682-685.
[15]欧阳臻,陈钧.不同季节桑叶中1-脱氧野尻霉素(DNJ)含量的测定.食品科学,2004,25(10):211-214.OUYANG Z,CHEN J.Determination of 1-deoxynojirimycin in Morus alba L.leaves in different seasons.Food Science,2004,25(10):211-214.
[16]曾锐,祝勇军,刘超.HPLC-ELSD法测定桑树不同部位中的1-脱氧野尻霉素.华西药学杂志,2009,24(2):171-172.ZENG R,ZHU Y J,LIU C.Determination of 1-deoxynojinimycin in different parts of mulberry by HPLC-ELSD.West China Journal of Pharmaceutical Sciences,2009,24(2):171-172.
[17]姜永涛,陈继永,刘珂.HPLC-ELSD测定桑叶中1-脱氧野尻霉素的含量.药物分析杂志,2005,25(1):27-29.JIANG Y T,CHEN J Y,LIU K.HPLC-ELSD determination of 1-deoxynojinimycin in mulberry leaves.Chinese Journal of Pharmaceutical Analysis,2005,25(1):27-29.
[18]肖洪,黄先智,沈以红,丁晓雯,张亚琼,梁菡峪,张迪.不同发酵条件对发酵桑叶茶生物活性成分含量的影响.食品科学,2013,34(23):216-220.XIAO H,HUANG X Z,SHEN Y H,DING X W,ZHANG Y Q,LIANG H Y,ZHANG D.Effects of different fermentation condition on the contents of bioactive ingredients in fermented mulberry leaf tea.Food Science,2013,34(23):216-220.
[19]胡瑞君,车振明.微波萃取桑叶1-脱氧野尻霉素工艺的研究.食品工程,2008(1):50-53.HU R J,CHE Z M.Studies on the microwave-assisted extraction method of DNJ from mulberry leaves.Food Engineering,2008(1):50-53.
[20]郑晓静,伍金娥,何计国,高月.响应面法优化超声波提取桑叶中1-脱氧野尻霉素的工艺研究.中国食品学报,2012,12(9):34-40.ZHENG X J,WU J E,HE J G,GAO Y.Optimization of 1-deoxynojirimycin extraction from mulberry leaves by using response surface methodology.Journal of Chinese Institute of Food Science and Technology,2012,12(9):34-40.
[21]刘凡,李平平,廖森泰,肖更生,邹宇晓,施英,吴娱明,唐翠明.98份不同桑树品种资源的桑叶总生物碱及1-脱氧野尻霉素含量测定.蚕业科学,2012,38(2):0185-0191.LIU F,LI P P,LIAO S T,XIAO G S,ZOU Y X,SHI Y,WU Y M,TANG C M.Determination of total alkaloids and 1-deoxynojirimycin contents in leaves of 98mulberry variety resources.Science of Sericulture,2012,38(2):0185-0191.
[22]刘伟鋆,赵骏,高秀梅,刘美平,曾森.1-脱氧野尻霉素提取精制工艺的研究.中华中医药学刊,2009,27(10):2079-2081.LIU W J,ZHAO J,GAO X M,LIU M P,ZENG S.Study on the extraction and purification of DNJ from Morus alba L.Chinese Archives of Traditional Chinese Medicine.2009,27(10):2079-2081.
[23]秦樱瑞,曾艺涛,杨娟,丁晓雯,黄先智.干燥方式对桑叶降糖活性成分含量的影响.食品科学,2015,36(17):71-76.QIN Y R,ZENG Y T,YANG J,DING X W,HUANG X Z.Effects of drying methods on the contents of hypoglycemic components in mulberry leaves(Morus alba L.).Food Science,2015,36(17):71-76.
[24]刘红英,何军,何珺.不同干燥方法对桑白皮中DNJ含量的影响.中药材,2012,35(7):1053-1055.LIU H Y,HE J,HE J.Effect of different drying methods on DNJ content in Cortex Mori.Journal of Chinese Medicinal Materials,2012,35(7):1053-1055.