沙棘粕醇提物的定性定量分析及其对衰老小鼠肝脏抗氧化指标的影响
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摘要
利用沙棘粕为研究对象,经乙醇提取、大孔吸附树脂纯化获得沙棘粕醇提物(sea buckthorn seed extract,SBSE)。利用超高效液相色谱-串联四极杆飞行时间质谱(ultra-performance liquid chromatography coupled with quadruple time-of-flight tandem mass spectrometry,UPLC-QTOF-MS/MS)对SBSE进行定性研究,通过超高效液相色谱-串联三重四极杆质谱(ultra performance liquid chromatography coupled with triple quadruple mass spectrometry,UPLC-QQQ-MS)定量分析SBSE活性成分。最后研究分析SBSE对以D-半乳糖诱导的衰老小鼠的抗氧化功效,不同浓度SBSE灌胃衰老小鼠模型42 d后,比较小鼠肝脏组织中总抗氧化能力(total antioxidant capacity,T-AOC)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性以及丙二醛(malondialdehyde,MDA)和脂褐素(lipofuscin,LP)水平。结果表明:通过UPLC-QTOF-MS/MS分析方法共鉴定出24种化合物,其中与标准品比对共确认14种物质;利用UPLC-QQQ-MS法,采用负离子多反应监测方式,建立SBSE的14种化合物的定量分析方法,通过检测分析,SBSE中山柰酚及其衍生物所占比例最大,质量浓度为(667.94±5.61)μg/mL;其次为槲皮素及其衍生物((204.01±0.04)μg/mL),原花青素类物质为(33.97±0.49)μg/mL。经过42 d的衰老小鼠灌胃实验,与模型组相比,灌胃一定剂量SBSE可以提高小鼠肝脏组织中GSH-Px、SOD和CAT的活力;低剂量组小鼠肝脏组织中的T-AOC水平极显著高于模型组(P<0.01);3种灌胃剂量小鼠的肝脏组织LP和MDA含量均极显著低于模型组(P<0.01)。
This research studied the qualitative analysis of sea buckthorn seed extract (SBSE), prepared from sea buckthorn seed residue by ethanol extraction and macroporous resin purification, by ultra-performance liquid chromatography coupled with quadruple time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). The bioactive components of SBSE were quantitatively determined by ultra performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS). Meanwhile, the antioxidant effect of SBSE in D-galactose-induced aging mice was studied. The mice were administrated by gavage with SBSE at different dosages for 42 days. Total antioxidant capacity (T-AOC), the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) and the contents of malondialdehyde (MDA) and lipofuscin (LP) in liver tissue were measured. The results obtained were as follows: A total of 24 compounds were identified by UPLC-QTOF-MS/MS, 14 compounds of which were characterized by comparing with reference substances and quantitatively determined by UPLC-QQQ-MS in the negative ion multiple reaction monitoring (MRM) mode. Kaempferol and its derivatives were the most abundant constituents in SBSE ((667.94 ± 5.61) μg/mL), quercetin and its derivatives were in the middle ((204.01 ± 0.04) μg/m L), and procyanidins were the least abundant ((33.97 ± 0.49) μg/m L). The administration of SBSE to aging mice could increase the activities of GSH-Px, SOD and CAT in liver tissue. Compared with the D-galactose-induced model group, hepatic T-AOC activity in the low-dosage group was significantly increased (P < 0.01), and the levels of MDA and LP were decreased significantly by SBSE at three dosages (P < 0.01).
This research studied the qualitative analysis of sea buckthorn seed extract (SBSE), prepared from sea buckthorn seed residue by ethanol extraction and macroporous resin purification, by ultra-performance liquid chromatography coupled with quadruple time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). The bioactive components of SBSE were quantitatively determined by ultra performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS). Meanwhile, the antioxidant effect of SBSE in D-galactose-induced aging mice was studied. The mice were administrated by gavage with SBSE at different dosages for 42 days. Total antioxidant capacity (T-AOC), the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) and the contents of malondialdehyde (MDA) and lipofuscin (LP) in liver tissue were measured. The results obtained were as follows: A total of 24 compounds were identified by UPLC-QTOF-MS/MS, 14 compounds of which were characterized by comparing with reference substances and quantitatively determined by UPLC-QQQ-MS in the negative ion multiple reaction monitoring (MRM) mode. Kaempferol and its derivatives were the most abundant constituents in SBSE ((667.94 ± 5.61) μg/mL), quercetin and its derivatives were in the middle ((204.01 ± 0.04) μg/m L), and procyanidins were the least abundant ((33.97 ± 0.49) μg/m L). The administration of SBSE to aging mice could increase the activities of GSH-Px, SOD and CAT in liver tissue. Compared with the D-galactose-induced model group, hepatic T-AOC activity in the low-dosage group was significantly increased (P < 0.01), and the levels of MDA and LP were decreased significantly by SBSE at three dosages (P < 0.01).
引文
[1]URSACHE F M,GHINEA I O,TURTURICA M,et al.Phytochemicals content and antioxidant properties of sea buckthorn (Hippophae rhamnoides L.) as affected by heat treatment-quantitative spectroscopic and kinetic approaches[J].Food Chemistry,2017,233:442-449.DOI:10.1016/j.foodchem.2017.04.107.
[2]ZHANG W,ZHANG X,ZOU K,et al.Seabuckthorn berry polysaccharide protects against carbon tetrachloride-induced hepatotoxicity in mice via anti-oxidative and anti-inflammatory activities[J].Food and Function,2017,8(9):3130-3138.DOI:10.1039/c7fo00399d.
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[5]ZIELINSKA A,NOWAK I.Abundance of active ingredients in sea-buckthorn oil[J].Lipids Health Disease,2017,16(1):95-105.DOI:10.1186/s12944-017-0469-7.
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[7]OLAS B.Sea buckthorn as a source of important bioactive compounds in cardiovascular diseases[J].Food and Chemical Toxicology,2016,97:199-204.DOI:10.1016/j.fct.2016.09.008.
[8]赖国银,王俐娟,卢鹤,等.超高效液相色谱-四极杆-飞行时间质谱定性研究茶叶籽中的酚类化合物[J].色谱,2017,35(5):502-508.DOI:10.3724/SP.J.1123.2016.11010.
[9]PéREZ-JIMéNEZ J,TORRES J L.Analysis of proanthocyanidins in almond blanch water by HPLC-ESI-QqQ-MS/MS and MALDI-TOF/TOF MS[J].Food Research International,2012,49(2):798-806.DOI:10.1016/j.foodres.2012.09.005.
[10]赵佳丽,徐盼盼,陈娟娟,等.基于HPLC-QqQ-MS/MS技术的坛紫菜中植物激素分析[J].食品科学,2016,37(16):216-222.DOI:10.7506/spkx1002-6630-201616035.
[11]张佳婵,王昌涛,刘瑶,等.沙棘粕醇提物对秀丽隐杆线虫的抗衰老功效及其机制的研究[J].食品科学,2017,38(23):141-148.DOI:10.7506/spkx1002-6630-201723023.
[12]张佳婵,史豆豆,王昌涛,等.细胞水平评价沙棘粕醇提物的抗衰老功效[J].食品科学,2017,38(19):164-170.DOI:10.7506/spkx1002-6630-201719026.
[13]ZHUANG X,ZHANG W,PANG X,et al.Combined effect of total flavonoids from seed residues of Hippophae rhamnoides L.and zinc on advanced glycation end products-induced endothelial cell dysfunction[J].Food Chemistry,2012,133(3):905-911.DOI:10.1016/j.foodchem.2012.02.001.
[14]VARSHNEYA C,KANT V,MEHTA M.Total phenolic contents and free radical scavenging activities of different extracts of seabuckthorn (Hippophae rhamnoides) pomace without seeds[J].International Journal of Food Sciences and Nutrition,2012,63(2):153-159.DOI:10.3109/09637486.2011.608652.
[15]ARIMBOOR R,ARUMUGHAN C.HPLC-DAD-MS/MS profiling of antioxidant flavonoid glycosides in sea buckthorn (Hippophae rhamnoides L.) seeds[J].International Journal of Food Sciences and Nutrition,2012,63(6):730-738.DOI:10.3109/09637486.2011.652075.
[16]ZHANG W,ZHAO J,ZHU X,et al.Antihyperglycemic effect of aqueous extract of sea buckthorn (Hippophae rhamnoides L.) seed residues in streptozotocin-treated and high fat-diet-fed rats[J].Journal of Food Biochemistry,2010,34(4):856-868.DOI:10.1111/j.1745-4514.2010.00337.x.
[17]DHYANI D,MAIKHURI R K,RAO K S,et al.Basic nutritional attributes of Hippophae rhamnoides (sea buckthorn) populations from Uttarakhand Himalaya,India[J].Current Science,2007,92(8):1148-1152.
[18]RANJITH A,KUMAR K S,VENUGOPALAN V V,et al.Fatty acids,tocols,and carotenoids in pulp oil of three sea buckthorn species(Hippophae rhamnoides,H.salicifolia,and H.tibetana)grown in the Indian Himalayas[J].Journal of the American Oil Chemists’Society,2006,83(4):359-364.DOI:10.1007/s11746-006-1213-z.
[19]CHO C H,JANG H,LEE M,et al.Sea buckthorn (Hippophae rhamnoides L.) leaf extracts protect neuronal PC-12 cells from oxidative stress[J].Journal of Microbiology and Biotechnology,2017,27(7):1257-1265.DOI:10.4014/jmb.1704.04033.
[20]陈玮玲,钟培培,王远兴.青钱柳叶活性成分的抗氧化活性及UPLC-QTOF-MS-MS分析[J].食品科学,2017,38(8):122-128.DOI:10.7506/spkx1002-6630-201708020.
[21]RIHAM O B,REHAM W,NOHA S,et al.Characterization of the bioactive constituents of Nymphaea alba rhizomes and evaluation of anti-biofilm as well as antioxidant and cytotoxic properties[J].Journal of Medicinal Plant Research,2016,10(26):390-401.DOI:10.5897/JMPR2016.6162.
[22]OLSOVSKA J,KAMENIK Z,CEJKA P,et al.Ultra-high-performance liquid chromatography profiling method for chemical screening of proanthocyanidins in Czech hops[J].Talanta,2013,116:919-926.DOI:10.1016/j.talanta.2013.07.067.
[23]FRACASSETTI D,COSTA C,MOULAY L,et al.Ellagic acid derivatives,ellagitannins,proanthocyanidins and other phenolics,vitamin C and antioxidant capacity of two powder products from camu-camu fruit(Myrciaria dubia)[J].Food Chemistry,2013,139(1/2/3/4):578-588.DOI:10.1016/j.foodchem.2013.01.121.
[24]SARNOSKI P J,JOHNSON J V,REED K A,et al.Separation and characterisation of proanthocyanidins in Virginia type peanut skins by LC-MSn[J].Food Chemistry,2012,131(3):927-939.DOI:10.1016/j.foodchem.2011.09.081.
[25]JIN A,OZGA J A,LOPES-LUTZ D,et al.Characterization of proanthocyanidins in pea (Pisum sativum L.),lentil (Lens culinaris L.),and faba bean (Vicia faba L.) seeds[J].Food Research International,2012,46(2):528-535.DOI:10.1016/j.foodres.2011.11.018.
[26]许文,傅志勤,林婧,等.HPLC-Q-TOF-MS和UPLC-QqQ-MS的三叶青主要成分定性与定量研究[J].中国中药杂志,2014,39(22):4365-4371.
[27]YANG H,QU Z,ZHANG J,et al.Ferulic acid ameliorates memory impairment in D-galactose-induced aging mouse model[J].International Journal of Food Sciences and Nutrition,2016,67(7):806-817.DOI:10.1080/09637486.2016.1198890.
[28]DE MAGALH?ES J P,STEVENS M,THORNTON D.The business of anti-aging science[J].Trends in Biotechnology,2017,35(11):1062-1073.DOI:10.1016/j.tibtech.2017.07.004.
[29]原慧萍,杨泽.氧化应激与衰老研究进展[J].中国老年保健医,2015,13(5):14-17.DOI:10.3969/j.issn.1672-4860.2015.05.004.
[30]RODRIGO R,BOSCO C.Oxidative stress and protective effects of polyphenols:comparative studies in human and rodent kidney.Αreview[J].Comparative Biochemistry and Physiology:Part C,2006,142(3):317-327.DOI:10.1016/j.cbpc.2005.11.002.
[31]DUQUE L,BRAVO K,OSORIO E.A holistic anti-aging approach applied in selected cultivated medicinal plants:a view of photoprotection of the skin by different mechanisms[J].Industrial Crops and Products,2017,97:431-439.DOI:10.1016/j.indcrop.2016.12.059.
[32]VAISERMAN A M,LUSHCHAK O V,KOLIADA A K.Anti-aging pharmacology:promises and pitfalls[J].Ageing Research Reviews,2016,31:9-35.DOI:10.1016/j.arr.2016.08.004.
[33]LARA J,SHERRATT M J,REES M.Aging and anti-aging[J].Maturitas,2016,93:1-3.DOI:10.1016/j.maturitas.2016.08.020.
[34]ANAND K V,MOHAMED J M S,THOMAS P A,et al.Protective role of chrysin against oxidative stress in D-galactose-induced aging in an experimental rat model[J].Geriatrics Gerontology International,2012,12(4):741-750.DOI:10.1111/j.1447-0594.2012.00843.x.
[35]DU Z,YANG Q,ZHOU T,et al.D-galactose-induced mitochondrial DNA oxidative damage in the auditory cortex of rats[J].Molecular Medicine Reports,2014,10(6):2861-2867.DOI:10.3892/mmr.2014.2653.
[2]ZHANG W,ZHANG X,ZOU K,et al.Seabuckthorn berry polysaccharide protects against carbon tetrachloride-induced hepatotoxicity in mice via anti-oxidative and anti-inflammatory activities[J].Food and Function,2017,8(9):3130-3138.DOI:10.1039/c7fo00399d.
[3]WAGH R V,CHATLI M K.Response surface optimization of extraction protocols to obtain phenolic rich antioxidant from sea buckthorn and their potential application into model meat system[J].Journal of Food Science and Technology,2017,54(6):1565-1576.DOI:10.1007/s13197-017-2588-6.
[4]ZHOU W,YUAN Z,LI G,et al.Isolation and structure determination of a new flavone glycoside from seed residues of seabuckthorn (Hippophae rhamnoides L.)[J].Natural Product Research,2017,32(8):1-6.DOI:10.1080/14786419.2017.1366481.
[5]ZIELINSKA A,NOWAK I.Abundance of active ingredients in sea-buckthorn oil[J].Lipids Health Disease,2017,16(1):95-105.DOI:10.1186/s12944-017-0469-7.
[6]WANI T A,WANI S M,AHMAD M,et al.Bioactive profile,health benefits and safety evaluation of sea buckthorn (Hippophae rhamnoides L.):a review[J].Cogent Food and Agriculture,2016,2(1):1128519.DOI:10.1080/23311932.2015.1128519.
[7]OLAS B.Sea buckthorn as a source of important bioactive compounds in cardiovascular diseases[J].Food and Chemical Toxicology,2016,97:199-204.DOI:10.1016/j.fct.2016.09.008.
[8]赖国银,王俐娟,卢鹤,等.超高效液相色谱-四极杆-飞行时间质谱定性研究茶叶籽中的酚类化合物[J].色谱,2017,35(5):502-508.DOI:10.3724/SP.J.1123.2016.11010.
[9]PéREZ-JIMéNEZ J,TORRES J L.Analysis of proanthocyanidins in almond blanch water by HPLC-ESI-QqQ-MS/MS and MALDI-TOF/TOF MS[J].Food Research International,2012,49(2):798-806.DOI:10.1016/j.foodres.2012.09.005.
[10]赵佳丽,徐盼盼,陈娟娟,等.基于HPLC-QqQ-MS/MS技术的坛紫菜中植物激素分析[J].食品科学,2016,37(16):216-222.DOI:10.7506/spkx1002-6630-201616035.
[11]张佳婵,王昌涛,刘瑶,等.沙棘粕醇提物对秀丽隐杆线虫的抗衰老功效及其机制的研究[J].食品科学,2017,38(23):141-148.DOI:10.7506/spkx1002-6630-201723023.
[12]张佳婵,史豆豆,王昌涛,等.细胞水平评价沙棘粕醇提物的抗衰老功效[J].食品科学,2017,38(19):164-170.DOI:10.7506/spkx1002-6630-201719026.
[13]ZHUANG X,ZHANG W,PANG X,et al.Combined effect of total flavonoids from seed residues of Hippophae rhamnoides L.and zinc on advanced glycation end products-induced endothelial cell dysfunction[J].Food Chemistry,2012,133(3):905-911.DOI:10.1016/j.foodchem.2012.02.001.
[14]VARSHNEYA C,KANT V,MEHTA M.Total phenolic contents and free radical scavenging activities of different extracts of seabuckthorn (Hippophae rhamnoides) pomace without seeds[J].International Journal of Food Sciences and Nutrition,2012,63(2):153-159.DOI:10.3109/09637486.2011.608652.
[15]ARIMBOOR R,ARUMUGHAN C.HPLC-DAD-MS/MS profiling of antioxidant flavonoid glycosides in sea buckthorn (Hippophae rhamnoides L.) seeds[J].International Journal of Food Sciences and Nutrition,2012,63(6):730-738.DOI:10.3109/09637486.2011.652075.
[16]ZHANG W,ZHAO J,ZHU X,et al.Antihyperglycemic effect of aqueous extract of sea buckthorn (Hippophae rhamnoides L.) seed residues in streptozotocin-treated and high fat-diet-fed rats[J].Journal of Food Biochemistry,2010,34(4):856-868.DOI:10.1111/j.1745-4514.2010.00337.x.
[17]DHYANI D,MAIKHURI R K,RAO K S,et al.Basic nutritional attributes of Hippophae rhamnoides (sea buckthorn) populations from Uttarakhand Himalaya,India[J].Current Science,2007,92(8):1148-1152.
[18]RANJITH A,KUMAR K S,VENUGOPALAN V V,et al.Fatty acids,tocols,and carotenoids in pulp oil of three sea buckthorn species(Hippophae rhamnoides,H.salicifolia,and H.tibetana)grown in the Indian Himalayas[J].Journal of the American Oil Chemists’Society,2006,83(4):359-364.DOI:10.1007/s11746-006-1213-z.
[19]CHO C H,JANG H,LEE M,et al.Sea buckthorn (Hippophae rhamnoides L.) leaf extracts protect neuronal PC-12 cells from oxidative stress[J].Journal of Microbiology and Biotechnology,2017,27(7):1257-1265.DOI:10.4014/jmb.1704.04033.
[20]陈玮玲,钟培培,王远兴.青钱柳叶活性成分的抗氧化活性及UPLC-QTOF-MS-MS分析[J].食品科学,2017,38(8):122-128.DOI:10.7506/spkx1002-6630-201708020.
[21]RIHAM O B,REHAM W,NOHA S,et al.Characterization of the bioactive constituents of Nymphaea alba rhizomes and evaluation of anti-biofilm as well as antioxidant and cytotoxic properties[J].Journal of Medicinal Plant Research,2016,10(26):390-401.DOI:10.5897/JMPR2016.6162.
[22]OLSOVSKA J,KAMENIK Z,CEJKA P,et al.Ultra-high-performance liquid chromatography profiling method for chemical screening of proanthocyanidins in Czech hops[J].Talanta,2013,116:919-926.DOI:10.1016/j.talanta.2013.07.067.
[23]FRACASSETTI D,COSTA C,MOULAY L,et al.Ellagic acid derivatives,ellagitannins,proanthocyanidins and other phenolics,vitamin C and antioxidant capacity of two powder products from camu-camu fruit(Myrciaria dubia)[J].Food Chemistry,2013,139(1/2/3/4):578-588.DOI:10.1016/j.foodchem.2013.01.121.
[24]SARNOSKI P J,JOHNSON J V,REED K A,et al.Separation and characterisation of proanthocyanidins in Virginia type peanut skins by LC-MSn[J].Food Chemistry,2012,131(3):927-939.DOI:10.1016/j.foodchem.2011.09.081.
[25]JIN A,OZGA J A,LOPES-LUTZ D,et al.Characterization of proanthocyanidins in pea (Pisum sativum L.),lentil (Lens culinaris L.),and faba bean (Vicia faba L.) seeds[J].Food Research International,2012,46(2):528-535.DOI:10.1016/j.foodres.2011.11.018.
[26]许文,傅志勤,林婧,等.HPLC-Q-TOF-MS和UPLC-QqQ-MS的三叶青主要成分定性与定量研究[J].中国中药杂志,2014,39(22):4365-4371.
[27]YANG H,QU Z,ZHANG J,et al.Ferulic acid ameliorates memory impairment in D-galactose-induced aging mouse model[J].International Journal of Food Sciences and Nutrition,2016,67(7):806-817.DOI:10.1080/09637486.2016.1198890.
[28]DE MAGALH?ES J P,STEVENS M,THORNTON D.The business of anti-aging science[J].Trends in Biotechnology,2017,35(11):1062-1073.DOI:10.1016/j.tibtech.2017.07.004.
[29]原慧萍,杨泽.氧化应激与衰老研究进展[J].中国老年保健医,2015,13(5):14-17.DOI:10.3969/j.issn.1672-4860.2015.05.004.
[30]RODRIGO R,BOSCO C.Oxidative stress and protective effects of polyphenols:comparative studies in human and rodent kidney.Αreview[J].Comparative Biochemistry and Physiology:Part C,2006,142(3):317-327.DOI:10.1016/j.cbpc.2005.11.002.
[31]DUQUE L,BRAVO K,OSORIO E.A holistic anti-aging approach applied in selected cultivated medicinal plants:a view of photoprotection of the skin by different mechanisms[J].Industrial Crops and Products,2017,97:431-439.DOI:10.1016/j.indcrop.2016.12.059.
[32]VAISERMAN A M,LUSHCHAK O V,KOLIADA A K.Anti-aging pharmacology:promises and pitfalls[J].Ageing Research Reviews,2016,31:9-35.DOI:10.1016/j.arr.2016.08.004.
[33]LARA J,SHERRATT M J,REES M.Aging and anti-aging[J].Maturitas,2016,93:1-3.DOI:10.1016/j.maturitas.2016.08.020.
[34]ANAND K V,MOHAMED J M S,THOMAS P A,et al.Protective role of chrysin against oxidative stress in D-galactose-induced aging in an experimental rat model[J].Geriatrics Gerontology International,2012,12(4):741-750.DOI:10.1111/j.1447-0594.2012.00843.x.
[35]DU Z,YANG Q,ZHOU T,et al.D-galactose-induced mitochondrial DNA oxidative damage in the auditory cortex of rats[J].Molecular Medicine Reports,2014,10(6):2861-2867.DOI:10.3892/mmr.2014.2653.