不同处理方法对花椒籽种皮黑色素纯化效果的比较研究
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摘要
本文以花椒籽为原料制备花椒籽种皮黑色素粗提物(melanin crude extract of pepper seed coat,MCEP),并用盐酸水解法、酶解法、有机溶剂萃取法和碱溶酸沉法四种处理方法对MCEP进行纯化,制得花椒籽种皮黑色素(melanin of pepper seed coat,MP),分别标记为SMP、EMP、OMP和AMP;通过分析其色值、紫外-可见光谱、色价、红外光谱及微观结构,对比研究了四种处理方法对MP的纯化效果。结果表明,SMP的L*、a*与b*值均显著低于AMP、OMP和MCEP(p<0.05),且为黑色无定型粉末,并带有金属光泽;四种方法制备的MP紫外-可见光谱均在220 nm有最大吸收峰,且色价均显著高于MCEP(151.25 U/g)(p<0.05),其中SMP色价(341.73 U/g)最高;除AMP外,OMP、EMP和SMP的红外光谱相比于MCEP在C-H与C-H2处的吸收峰均明显减弱;扫描电子显微镜结果显示,SMP呈无规则块状,且表面无其他物质形状结构存在。因此,盐酸水解为本研究纯化制备MP的最佳处理方法。
Pepper seeds was used as raw material to prepare crude melanin extract of pepper seed coat(MCEP) by alkali-soluble acid precipitation method, and the MCEP was purified by hydrochloric acid hydrolysis, enzymatic hydrolysis, organic solvent extraction and alkali-soluble acid precipitation method, respectively, to obtain melanins of pepper seed coat(MP), which were labeled as SMP, EMP, OMP and AMP, respectively. The purification effects of the four treatment methods on MP were analyzed by the color of the pigment, ultraviolet-visible spectroscopy, color value, Infrared spectroscopy and its microstructure. The results showed that the L*, a* and b* values of SMP were significantly lower than those of AMP, OMP and MCEP(p<0.05), and the SMP was black amorphous powder with metallic luster; The SMP,EMP, OMP and AMP had maximum absorption at peak 220 nm; the color values of SMP, EMP, OMP and AMP were significantly higher than the color value of MCEP(151.25 U/g)(p<0.05), and the SMP color value(341.73 U/g) was the highest; Except for AMP, the infrared spectroscopy of OMP, EMP and SMP were weakened or disappeared compared with the absorption peaks of MCEP in C-H and C-H2 characteristic absorption band; The results of scanning electron microscope showed that SMP had a random block structure, and no other structural shapes existed. Therefore, we suggest that the hydrochloric acid hydrolysis is the best treatment method for purification and preparation of MP.
Pepper seeds was used as raw material to prepare crude melanin extract of pepper seed coat(MCEP) by alkali-soluble acid precipitation method, and the MCEP was purified by hydrochloric acid hydrolysis, enzymatic hydrolysis, organic solvent extraction and alkali-soluble acid precipitation method, respectively, to obtain melanins of pepper seed coat(MP), which were labeled as SMP, EMP, OMP and AMP, respectively. The purification effects of the four treatment methods on MP were analyzed by the color of the pigment, ultraviolet-visible spectroscopy, color value, Infrared spectroscopy and its microstructure. The results showed that the L*, a* and b* values of SMP were significantly lower than those of AMP, OMP and MCEP(p<0.05), and the SMP was black amorphous powder with metallic luster; The SMP,EMP, OMP and AMP had maximum absorption at peak 220 nm; the color values of SMP, EMP, OMP and AMP were significantly higher than the color value of MCEP(151.25 U/g)(p<0.05), and the SMP color value(341.73 U/g) was the highest; Except for AMP, the infrared spectroscopy of OMP, EMP and SMP were weakened or disappeared compared with the absorption peaks of MCEP in C-H and C-H2 characteristic absorption band; The results of scanning electron microscope showed that SMP had a random block structure, and no other structural shapes existed. Therefore, we suggest that the hydrochloric acid hydrolysis is the best treatment method for purification and preparation of MP.
引文
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[19]张伟,吴博.花椒籽黑色素化学结构的初步研究[J].中国测试,2017,43(3):49-52ZHANG Wei,WU Bo.Study on structure characteristics of Chinese prickly ash melanin[J].China Measurement&Test,2017,43(3):49-52
[20]姚增玉.山杏种皮黑色素提取及其性质研究[D].杨凌:西北农林科技大学,2007YAO Zeng-yu.Isolation and characterization of melanin from testae of wild apricots[D].Yangling:Northwest A&FUniversity,2007
[21]SUN S,ZHANG X,SUN S,et al.Production of natural melanin by Auricularia auricula and study on its molecular structure[J].Food Chemistry,2016,190:801-807
[22]XIN C,MA J,TAN C,et al.Preparation of melanin from Catharsius molossus L.and preliminary study on its chemical structure[J].Journal of Bioscience&Bioengineering,2015,119(4):446-454
[23]罗强,杨雪果,施宝珠,等.栓皮栎橡子壳色素理化性质和抗氧化活性[J].食品科学,2017,38(11):128-134LUO Qiang,YANG Xue-guo,SHI Bao-zhu,et al.Physicochemical properties and antioxidant activity of Quercus variabilis acorn shell pigment[J].Food Science,2017,38(11):128-134
[24]邹宇,尹冬梅,冮洁,等.黑木耳黑色素组分分析及其抗氧化活性研究[J].食品科学,2013,34(23):138-141ZOU Yu,YIN Dong-mei,GANG Jie,et al.Composition analysis and antioxidant activity of Auricularia auricula melanin[J].Food Science,2013,34(23):138-141
[25]康静静,胡海威,温文,等.响应面法优化黑芝麻黑色素的碱提条件[J].中国食品添加剂,2014,4:92-97KANG Jing-jing,HU Hai-wei,WEN Wen,et al.Optimization of black pigment alkaline extraction from black sesame by response surface methodology[J].China Food Additives,2014,4:92-97
[26]HUANG S,PAN Y,GAN D,et al.Antioxidant activities and UV-protective properties of melanin from the berry of Cinnamomum burmannii and Osmanthus fragrans[J].Medicinal Chemistry Research,2011,20(4):475-481
[27]张小林,陈博文,孙晓瑞,等.响应面法优化超声波辅助灵芝黑色素提取工艺[J].核农学报,2018,32(3):532-538ZHANG Xiao-lin,CHEN Bo-wen,SUN Xiao-rui,et al.The response surface method to optimize the ultrasonic-assisted extraction of Ganoderma lucidum melanin[J].Journal of Nuclear Agricultural Sciences,2018,32(3):532-538
[28]Selvakumar P,Rajasekar S,Periasamy K,et al.Isolation and characterization of melanin pigment from Pleurotus cystidiosus(telomorph of Antromycopsis macrocarpa)[J].World Journal of Microbiology&Biotechnology,2008,24(10):2125-2131
[29]SUN S,ZHANG X,CHEN W,et al.Production of natural edible melanin by Auricularia auricula and its physicochemical properties[J].Food Chemistry,2016,196:486-492
[30]WANG H S,PAN Y M,TANG X J,et al.Isolation and characterization of melanin from Osmanthus fragrans’seeds[J].LWT-Food Science and Technology,2006,39(5):496-502
[31]康静静.黑芝麻黑色素的分离及其结构性质的研究[D].南昌:南昌大学,2015KANG Jing-jing.Study of the extraction,purification,structural characteristics and physicochemical property of black pigment from semen sesami nigrum[D].Nanchang:Nanchang University,2015
[32]刘显威,刘淑集,张学勤,等.响应面法优化鱿鱼墨黑色素的制备工艺[J].海南大学学报(自然科学版),2016,34(1):25-32LIU Xian-wei,LIU Shu-ji,ZHANG Xue-qin,et al.Optimization of melanin preparation conditions from squid ink by response surface methodology[J].Natural Science Journal of Hainan University(Natural Science),2016,34(1):25-32
[33]YE M,CHEN X,LI G W,et al.Structural characteristics of pheomelanin-like pigment from Lachnum singerianum[J].Advanced Materials Research,2011,284-286:1742-1745
[34]Araujo M,Xavier J R,Nunes C D,et al.Marine sponge melanin:A new source of an old biopolymer[J].Structural Chemistry,2012,23(1):115-122
[2]Eriksen N T.Production of phycocyanin-a pigment with applications in biology,biotechnology,foods and medicine[J].Appl Microbiol Biotechnol,2008,80(1):1-14
[3]TAN M,GAN D,WEI L,et al.Isolation and characterization of pigment from Cinnamomum burmannii'peel[J].Food Research International,2011,44(7SI):2289-2294
[4]YAO Z Y,QI J H.Comparison of antioxidant activities of melanin fractions from Chestnut Shell[J].Molecules,2016,21(4):487
[5]董小华,课敬璇,申光辉,等.花椒籽黑色素体外抗氧化能力研究[J].中国油脂,2016,41(8):66-69DONG Xiao-hua,KE Jing-xuan,SHEN Guang-hui,et al.In vitro antioxidant capacity of melanin in pepper seeds[J].China Oils and Fats,2016,41(8):66-69
[6]YE M,GUO G Y,LU Y,et al.Purification,structure and anti-radiation activity of melanin from Lachnum YM404[J].International Journal of Biological Macromolecules,2014,63:170-176
[7]SONG S,LI S,SU N,et al.Structural characterization,molecular modification and hepatoprotective effect of melanin from Lachnum YM226 on acute alcohol-induced liver injury in mice[J].Food&Function,2016,7(8):3617-3627
[8]LU Y,YE M,SONG S,et al.Isolation,Purification,and Anti-Aging Activity of Melanin from Lachnum singerianum[J].Applied Biochemistry&Biotechnology,2014,174(2):762-771
[9]ZHU H,HE C C,CHU Q H.Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from Auricularia auricular[J].Letters in Applied Microbiology,2011,52(3):269-274
[10]郑淑英.乌饭树叶黑色素抗氧化性及其着色能力初步研究[J].宁德师范学院学报(自然科学版),2016,28(2):169-173ZHENG Shu-ying.Preliminary study on the antioxidant activity and coloring ability of melanin in leaves of wufan[J].Journal of Ningde Normal University(Natural Science Edition),2016,28(2):169-173
[11]刘晓娜,蒋慧兰,林广宇,等.香蕉皮中黑色素的提取及其对不同油脂的抗氧化作用研究[J].现代食品,2018,8:128-131LIN Xiao-na,JIANG Hui-lan,LIN Guang-yu,et al.Study on extraction of banana peel melanin and its antioxidantive effects on different edible oils[J].Modern Food,2018,8:128-131
[12]马萍,郭希娟,郭增旺.黑米黑色素微波辅助提取工艺优化[J].食品与机械,2014,30(5):229-231MA Ping,GUO Xi-juan,GUO Zeng-wang.Optimization on microwave-assisted extraction for melanin from black kerneled rice via quadratic regressive currency rotary design[J].Food&Machinery,2014,30(5):229-231
[13]任廷远.花椒麻素对试验大鼠蛋白质合成与分解代谢影响的机制研究[D].重庆:西南大学,2017REN Ting-yuan.The effects and mechanisms of Zanthoxylum alkylamides on protein synthesis and catabolism in rats[D].Chongqing:Southwest University,2017
[14]杨沫,薛媛,任璐,等.不同粒度花椒籽黑种皮粉理化特性[J].食品科学,2018,39(9):47-52YANG Mo,XUE Yuan,REN Lu,et al.Physicochemical characteristics of black seed coat powders with different particle sizes of pepper(Zanthoxylum bungeanum)[J].Food Science,2018,39(9):47-52
[15]李荣,贺学林,徐怀德.花椒籽黑色素提取和脱蛋白技术研究[J].西北植物学报,2008,28(12):2558-2563LI Rong,HE Xue-lin,XU Huai-de.Melanin isolation from Zanthoxylum bungeanum seeds and protein removal[J].Acta Botanica Boreali-occidentalia Sinica,2008,28(12):2558-2563
[16]徐文秀,王鹏,王晓燕.正交法优化花椒籽黑色素提取工艺研究[J].食品研究与开发,2014,6:49-52XU Wen-xiu,WANG Peng,WANG Xiao-yan.Research on optimizing extraction method of prickly ash seed melanin by orthogonal array design tests[J].Food Research and Development,2014,6:49-52
[17]杨秀芳,王改利,马养民.花椒籽种皮黑色素的提取纯化工艺[J].贵州农业科学,2013,41(10):149-153YANG Xiu-fang,WANG Gai-li,MA Yang-min.Extraction and purification technology for melanin in seed coat of chinese prickly ash[J].Guizhou Agricultural Sciences,2013,41(10):149-153
[18]杨秀芳,王改利,马养民.花椒籽种皮黑色素理化性质和抗氧化活性的研究[J].河南工业大学学报(自然科学版),2014,35(1):82-86YANG Xiu-fang,WANG Gai-li,MA Yang-min.Research on physical and chemical properties and antioxidant activity of melanin in seed coat of Chinese red pepper[J].Journal of Henan University of Technology(Natural Science),2014,35(1):82-86
[19]张伟,吴博.花椒籽黑色素化学结构的初步研究[J].中国测试,2017,43(3):49-52ZHANG Wei,WU Bo.Study on structure characteristics of Chinese prickly ash melanin[J].China Measurement&Test,2017,43(3):49-52
[20]姚增玉.山杏种皮黑色素提取及其性质研究[D].杨凌:西北农林科技大学,2007YAO Zeng-yu.Isolation and characterization of melanin from testae of wild apricots[D].Yangling:Northwest A&FUniversity,2007
[21]SUN S,ZHANG X,SUN S,et al.Production of natural melanin by Auricularia auricula and study on its molecular structure[J].Food Chemistry,2016,190:801-807
[22]XIN C,MA J,TAN C,et al.Preparation of melanin from Catharsius molossus L.and preliminary study on its chemical structure[J].Journal of Bioscience&Bioengineering,2015,119(4):446-454
[23]罗强,杨雪果,施宝珠,等.栓皮栎橡子壳色素理化性质和抗氧化活性[J].食品科学,2017,38(11):128-134LUO Qiang,YANG Xue-guo,SHI Bao-zhu,et al.Physicochemical properties and antioxidant activity of Quercus variabilis acorn shell pigment[J].Food Science,2017,38(11):128-134
[24]邹宇,尹冬梅,冮洁,等.黑木耳黑色素组分分析及其抗氧化活性研究[J].食品科学,2013,34(23):138-141ZOU Yu,YIN Dong-mei,GANG Jie,et al.Composition analysis and antioxidant activity of Auricularia auricula melanin[J].Food Science,2013,34(23):138-141
[25]康静静,胡海威,温文,等.响应面法优化黑芝麻黑色素的碱提条件[J].中国食品添加剂,2014,4:92-97KANG Jing-jing,HU Hai-wei,WEN Wen,et al.Optimization of black pigment alkaline extraction from black sesame by response surface methodology[J].China Food Additives,2014,4:92-97
[26]HUANG S,PAN Y,GAN D,et al.Antioxidant activities and UV-protective properties of melanin from the berry of Cinnamomum burmannii and Osmanthus fragrans[J].Medicinal Chemistry Research,2011,20(4):475-481
[27]张小林,陈博文,孙晓瑞,等.响应面法优化超声波辅助灵芝黑色素提取工艺[J].核农学报,2018,32(3):532-538ZHANG Xiao-lin,CHEN Bo-wen,SUN Xiao-rui,et al.The response surface method to optimize the ultrasonic-assisted extraction of Ganoderma lucidum melanin[J].Journal of Nuclear Agricultural Sciences,2018,32(3):532-538
[28]Selvakumar P,Rajasekar S,Periasamy K,et al.Isolation and characterization of melanin pigment from Pleurotus cystidiosus(telomorph of Antromycopsis macrocarpa)[J].World Journal of Microbiology&Biotechnology,2008,24(10):2125-2131
[29]SUN S,ZHANG X,CHEN W,et al.Production of natural edible melanin by Auricularia auricula and its physicochemical properties[J].Food Chemistry,2016,196:486-492
[30]WANG H S,PAN Y M,TANG X J,et al.Isolation and characterization of melanin from Osmanthus fragrans’seeds[J].LWT-Food Science and Technology,2006,39(5):496-502
[31]康静静.黑芝麻黑色素的分离及其结构性质的研究[D].南昌:南昌大学,2015KANG Jing-jing.Study of the extraction,purification,structural characteristics and physicochemical property of black pigment from semen sesami nigrum[D].Nanchang:Nanchang University,2015
[32]刘显威,刘淑集,张学勤,等.响应面法优化鱿鱼墨黑色素的制备工艺[J].海南大学学报(自然科学版),2016,34(1):25-32LIU Xian-wei,LIU Shu-ji,ZHANG Xue-qin,et al.Optimization of melanin preparation conditions from squid ink by response surface methodology[J].Natural Science Journal of Hainan University(Natural Science),2016,34(1):25-32
[33]YE M,CHEN X,LI G W,et al.Structural characteristics of pheomelanin-like pigment from Lachnum singerianum[J].Advanced Materials Research,2011,284-286:1742-1745
[34]Araujo M,Xavier J R,Nunes C D,et al.Marine sponge melanin:A new source of an old biopolymer[J].Structural Chemistry,2012,23(1):115-122