亚麻籽粕木酚素的纯化及对DNA氧化损伤保护
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摘要
以亚麻籽粕为原料,采用回流法提取木酚素。粗提液浓缩后经D101型大孔吸附树脂分离,进而通过半制备高效液相纯化,以Megres C_(18)(30 mm×250 mm,10μm)为色谱柱,乙腈(A)-0.1%磷酸水(B)为流动相,梯度洗脱,流速35mL/min,检测波长280nm,柱温30℃,进样量2 mL,得到纯度98%的亚麻木酚素产品。探究亚麻木酚素对H_2O_2及H_2O_2紫外光解反应诱导质粒DNA氧化损伤的保护作用,结果表明其质量浓度为0.07~0.40μg/μL时,对抑制DNA氧化损伤具有显著的保护作用。
Extracted lignan from flaxseed meal by reflux method.The crude extract was preliminary purified by D101 macroporous adsorption resins which was followed by semi-preparation HPLC separation.The semi-preparative HPLC conditions were as follows:column was Megres C_(18)(30 mm×250 mm,10μm),mobile phase was a mixture of acetonitrile(A)-0.1% phosphoric acid water solution(B),gradient elution,flow rate was 35 mL/min,wavelength was 280 nm,column temperature was 30 ℃ and injection volume was 2 mL.In the conditions,98% purity flax lignans were obtained.Investigated the protecting activity of lignan against DNA damage that caused by UV photolysis and H_2O_2 treatment,it reveals that flax lignans has a significant effect on the protection of DNA oxidative damage while the concentration is 0.07~0.40μg/μL.
Extracted lignan from flaxseed meal by reflux method.The crude extract was preliminary purified by D101 macroporous adsorption resins which was followed by semi-preparation HPLC separation.The semi-preparative HPLC conditions were as follows:column was Megres C_(18)(30 mm×250 mm,10μm),mobile phase was a mixture of acetonitrile(A)-0.1% phosphoric acid water solution(B),gradient elution,flow rate was 35 mL/min,wavelength was 280 nm,column temperature was 30 ℃ and injection volume was 2 mL.In the conditions,98% purity flax lignans were obtained.Investigated the protecting activity of lignan against DNA damage that caused by UV photolysis and H_2O_2 treatment,it reveals that flax lignans has a significant effect on the protection of DNA oxidative damage while the concentration is 0.07~0.40μg/μL.
引文
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[22]张溢,孙培冬,陈桂冰,等.芡实多糖的提取、抗氧化活性及对质粒DNA氧化损伤防护作用的研究[J].食品工业科技,2015,36(11):122-126.
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[24]KUMAR V,LEMOS M,SHARMA M,et al.Antioxidant and DNA damage protecting activities of Eulophia nuda,Lindl[J].Free Radicals&Antioxidants,2013,3(2):55-60.
[25]李国栋,阎雪冬,童坦君,等.不同鼠龄小鼠脾细胞对过氧化氢诱导DNA损伤的修复能力[J].中国老年学,2011,31(4):599-601.
[26]PONNUSAMY S,SANDHIYA L,SENTHILKUMAR K.Reaction mechanism and kinetics of the degradation of terbacil initiated by oh radical-a theoretical study[J].Chemical Physics,2018,501:110-120.
[27]葛晓静,邓天昇,侯相林.亚麻木酚素的稳定性及其提取工艺[J].食品与发酵工业,2009,35(7):174-178.
[28]EDEL A L,ALIANI M,PIERCE G N.Stability of bioactives in flaxseed and flaxseed-fortified foods[J].Food Research International,2015,77:140-155.
[2]付亚琦.亚麻籽木酚素不同水解产物结构鉴定及活性研究[D].杭州:浙江工商大学,2014:1-4.
[3]TEPONNO R B,KUSARI S,SPITELLER M.Recent advances in research on lignans and neolignans[J].Natural Product Reports,2016,33(9):1 044.
[4]ZHANG Wen-bin,XU Shi-ying.Microwave-assisted extraction of secoisolariciresinol diglucoside from flaxseed hull[J].Journal of the Science of Food&Agriculture,2010,87(8):1 455-1 462.
[5]LOWCOCK E C,COTTERCHIO M,BOUCHER B A.Consumption of flaxseed,a rich source of lignans,is associated with reduced breast cancer risk[J].Cancer Causes&Control,2013,24(4):813-816.
[6]CALIGIURI S P B,EDEL A L,ALIANI M,et al.Flaxseed for hypertension:implications for blood pressure regulation[J].Current Hypertension Reports,2014,16(12):1-13.
[7]PALLA A H,KHAN N A,BASHIR S,et al.Pharmacological basis for the medicinal use of Linum usitatissimum(Flaxseed)in infectious and non-infectious diarrhea[J].Journal of Ethnopharmacology,2015,160:61-68.
[8]HANO C,RENOUARD S,MOLINIE R,et al.Flaxseed(Linum usitatissimum,L.)extract as well as secoisolariciresinol diglucoside and its mammalian derivatives are potent inhibitors ofα-amylase activity[J].Bioorganic&Medicinal Chemistry Letters,2013,23(10):3 007-3 012.
[9]周小洁,车向荣,于霏.亚麻籽及其饼粕的营养学和毒理学研究进展[J].饲料工业,2005,26(19):46-50.
[10]翟双双,杨琳.亚麻籽粕脱毒工艺及其在动物饲料中的应用[J].中国饲料,2014(15):31-34.
[11]PIHLAVA J M,HYVARINEN H,RYHANEN E L,et al.Process for isolating and purifying secoisolariciresinol diglcoside(sdg)from flaxseed:US,20040030108[P].2004-02-12.
[12]ANDREAS D,SASKIA H,PETER W.Isolation of the lignan secoisolariciresinol diglucoside from flaxseed(Linum usitatissimum L.)by high-speed counter-current chromatography[J].Journal of Chromatography A,2002,943(2):299-302.
[13]张文斌,许时婴.大孔吸附树脂法分离纯化亚麻木酚素的研究[J].食品工业科技,2006(10):96-98,102.
[14]郭丽兵,王蕾.常用大孔吸附树脂的主要参数和应用情况[J].中国现代中药,2006(4):26-32.
[15]XIE Xiu-man,SUN Wan-yang,HUANG Jing-yi,et al.Preparative high performance liquid chromatography-based multidimensional chromatography and its application in traditional Chinese medicine[J].Chinese Journal of Analytical Chemistry,2016,44(7):1 140-1 147.
[16]LU Teng,FINKEL T.Free radicals and senescence[J].Experimental Cell Research,2008,314(9):1 918.
[17]GUPTA R K,PATEl A K,SHAH N,et al.Oxidative stress and antioxidants in disease and cancer:a review[J].Asian Pacific Journal of Cancer Prevention Apjcp,2014,15(11):4 405-4 409.
[18]SMOLSKAITE L,VENSKUTONIS P R,TALOU T.Comprehensive evaluation of antioxidant and antimicrobial properties of different mushroom species[J].LWT-Food Science and Technology,2015,60(1):462-471.
[19]HE Lu-ping,NING Xue-lian,SUN Yue,et al.Research progress in the mechanisms of DNA damage response in cell senescence and carcinogenesis[J].Chinese Journal of Cancer Prevention&Treatment,2017,24(11):783-787.
[20]杨雪艳,邓利,林风,等.大孔吸附树脂纯化亚麻木酚素[J].生物加工过程,2016,14(5):12-16.
[21]杨宏志,李静.大孔树脂分离纯化亚麻木酚素工艺[J].中国食品学报,2015,15(7):139-146.
[22]张溢,孙培冬,陈桂冰,等.芡实多糖的提取、抗氧化活性及对质粒DNA氧化损伤防护作用的研究[J].食品工业科技,2015,36(11):122-126.
[23]KUMAR A,CHATTOPADHYAY S.DNA damage protecting activity and antioxidant potential of pudina extract[J].Food Chemistry,2007,100(4):1 377-1 384.
[24]KUMAR V,LEMOS M,SHARMA M,et al.Antioxidant and DNA damage protecting activities of Eulophia nuda,Lindl[J].Free Radicals&Antioxidants,2013,3(2):55-60.
[25]李国栋,阎雪冬,童坦君,等.不同鼠龄小鼠脾细胞对过氧化氢诱导DNA损伤的修复能力[J].中国老年学,2011,31(4):599-601.
[26]PONNUSAMY S,SANDHIYA L,SENTHILKUMAR K.Reaction mechanism and kinetics of the degradation of terbacil initiated by oh radical-a theoretical study[J].Chemical Physics,2018,501:110-120.
[27]葛晓静,邓天昇,侯相林.亚麻木酚素的稳定性及其提取工艺[J].食品与发酵工业,2009,35(7):174-178.
[28]EDEL A L,ALIANI M,PIERCE G N.Stability of bioactives in flaxseed and flaxseed-fortified foods[J].Food Research International,2015,77:140-155.