辣椒果实类胡萝卜素的提取与测定方法研究
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摘要
旨在探索和优化辣椒果实类胡萝卜素提取与高效液相色谱(HPLC)检测体系。以辣椒成熟果实为材料,采用单因素试验与L9(33)正交试验相结合的方法,对提取溶剂、料液比、提取温度和时间及材料预处理等5个因素进行探讨,优化辣椒果实类胡萝卜素提取工艺;进而对流动相种类、配比和流速等进行优化。结果表明:以二氯甲烷-乙醇(V二氯甲烷:V乙醇=1:1)为提取溶剂,在料液比为1:18,50℃恒温提取60 min,辣椒果实类胡萝卜素产率最高。利用L600-2高效液相色谱仪,在色谱条件为:流动相A为甲醇-乙腈(V甲醇:V乙腈=55:45),流动相B为100%甲基叔丁基醚,A:B为90:10等度洗脱,流速为1 mL/min,进样量为20μl时能较好地鉴定辣椒果实类胡萝卜素主要组分。本试验优化的辣椒果实类胡萝卜素提取和HPLC测定法,操作简便,提取率高,重复性好,为今后辣椒果实类胡萝卜素的提取和测定提供了参考。
The study aims to explore and optimize the extraction of carotenoids from pepper fruit and highperformance liquid chromatography(HPLC) detection system. Mature pepper fruits were used as materials, by using single factor experiment and L9(33) orthogonal test, the main five factors including extraction solvent, the ratio of solid to liquid, extraction temperature, time and pretreatment method to the raw materials were discussed. And then the types of mobile phase, ratio and flow fate were also optimized. The results showed that by using dichloromethane-ethyl alcohol(V: V = 1:1) as extraction solvent, the material to liquid ratio of 1:18,extraction temperature and time of 50℃ and 60 min, the yield of carotenoids was the highest. Using L600-2 HPLC, the main components of carotenoids in pepper fruit could be identified by the optimized chromatographic condition such as: methanol and acetonitrile as mobile phase A(V: V =55:45), 100% methyl tertiary butyl ether as mobile phase B and VA: VB=90: 10, 1.0 mL/min of flow rate and 25℃ of column temperature, 20 μL of sample quantity, respectively. This optimized carotenoids extraction method and HPLC determination method are easily operated, have high productivity and good repeatability, and can provide a reference for the extraction and determination of carotenoids from pepper fruit in the future.
The study aims to explore and optimize the extraction of carotenoids from pepper fruit and highperformance liquid chromatography(HPLC) detection system. Mature pepper fruits were used as materials, by using single factor experiment and L9(33) orthogonal test, the main five factors including extraction solvent, the ratio of solid to liquid, extraction temperature, time and pretreatment method to the raw materials were discussed. And then the types of mobile phase, ratio and flow fate were also optimized. The results showed that by using dichloromethane-ethyl alcohol(V: V = 1:1) as extraction solvent, the material to liquid ratio of 1:18,extraction temperature and time of 50℃ and 60 min, the yield of carotenoids was the highest. Using L600-2 HPLC, the main components of carotenoids in pepper fruit could be identified by the optimized chromatographic condition such as: methanol and acetonitrile as mobile phase A(V: V =55:45), 100% methyl tertiary butyl ether as mobile phase B and VA: VB=90: 10, 1.0 mL/min of flow rate and 25℃ of column temperature, 20 μL of sample quantity, respectively. This optimized carotenoids extraction method and HPLC determination method are easily operated, have high productivity and good repeatability, and can provide a reference for the extraction and determination of carotenoids from pepper fruit in the future.
引文
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[12]莫玉楠,张鲁刚,王国芳.橙色大白菜类胡萝卜素提取与测定方法研究[J].西北农林科技大学学报:自然科学版,2014(3):206-214.
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[14]贾关荣,赵文恩,申红山,等.西瓜中类胡萝卜素的柱色谱法分离分析[J].安徽农业科学,2008(20):8416-8418.
[15]储大燕.烟草中类胡萝卜素及其异构体的分析研究[D].合肥:中国科学技术大学,2009.
[16]冯作山,陈计峦,孙高峰,等.枸杞色素的提取及纯化技术[J].食品与发酵工业,2004(12):141-144.
[17]彭光华,李忠,张声华.薄层色谱法分离鉴定枸杞子中的类胡萝卜素[J].营养学报,1998(1):79-81.
[18]丁来欣,陈里.薄层色谱法分离测定辣椒红色素中的辣椒红[J].色谱,1995(4):295-296.
[19] Luengo E, Alvarez I, Raso J. Improving carotenoid extraction fromtomato waste by pulsed electric fields[J]. Front Nutr, 2014,1:12.
[20]张玲,郑婷婷,魏伟淋,等.两种不同肉色枇杷果实类胡萝卜素积累及合成相关基因的表达[J].园艺学报,2015(11):2153-2162.
[21]李小丽,梁远学,郜凌超,等.油棕果实不同发育时期类胡萝卜素的含量变化[J].华中农业大学学报,2015(1):23-27.
[22] Richins R D, Kilcrease J, Rodgriguez-Uribe L, et al. CarotenoidExtraction and Quantification from Capsicum annuum[J]. BioProtoc, 2014,4(19).
[23]张海涛,蒋文艳,淡瑞芳.胡萝卜粉中类胡萝卜素的提取工艺优化[J].甘肃农业大学学报,2012(5):139-142.
[24]白婕,韦燕春.金橘类胡萝卜素提取研究[J].大众商务,2009(18):289-290.
[25] Konings E J M, Roomans H H S. Evaluation and validation of anLC method for the analysis of carotenoids in vegetables and fruit[J]. Food Chemistry,1997,59(4):599-603.
[26]胡玉琴,郁继华,王小龙,等.辣椒叶片中类胡萝卜素组分分离及测定方法的研究[J].西北师范大学学报:自然科学版,2015(4):75-80.
[27]王鑫.红辣椒色素的提取、分离及光稳定性研究[D].无锡:江南大学,2005.
[28]戴雄泽,王利群,陈文超,等.辣椒果实发育过程中果色与类胡萝卜素的变化[J].中国农业科学,2009(11):4004-4011.
[29]董彩英.辣椒果实类胡萝卜素的积累特点及其与果色的关系研究[D].扬州:扬州大学,2009.
[2]白寿宁.超临界CO2萃取枸杞色素中β-胡萝卜素研究[J].包装与食品机械,2000(2):18-21.
[3]郝磊,张坤生.超临界CO2萃取玉米蛋白粉中类胡萝卜素的研究[J].食品研究与开发,2007(2):57-60.
[4]胡玉琴.辣椒叶片中类胡萝卜素组分分离及测定方法研究[D].兰州:甘肃农业大学,2014.
[5]袁旭红,李洋,徐雅琴.沙棘类胡萝卜素复合酶法提取及纯化的优化研究[J].食品科技,2010(6):195-198.
[6]吴青莹,宋莹莹,邢思熙,等.香蕉皮类胡萝卜素提取工艺条件的研究[J].中国食物与营养,2016(4):32-34.
[7]胡利强,郭晓伟,赵文恩,等.西瓜中类胡萝卜素的分离与分析[J].河南农业科学,2011(9):150-156.
[8]陶能国,张继红,张妙玲,等.微波法提取枸杞中类胡萝卜素的工艺研究[J].湘潭大学自然科学学报,2008(2):48-51.
[9]廖春燕,磨文龙.微波法提取橘皮中类胡萝卜素的工艺研究[J].安徽农业科学,2009(32):15981-15983.
[10]颜少宾,张妤艳,马瑞娟,等.桃果实类胡萝卜素测定方法的研究[J].果树学报,2012(6):1127-1133.
[11]严娟,蔡志翔,沈志军,等.黄肉桃果实中类胡萝卜素提取和测定方法研究[J].果树学报,2015(6):1267-1274.
[12]莫玉楠,张鲁刚,王国芳.橙色大白菜类胡萝卜素提取与测定方法研究[J].西北农林科技大学学报:自然科学版,2014(3):206-214.
[13] Ishida B K, Chapman M H. Carotenoid extraction from plants usinga novel, environmentally friendly solvent[J]. J Agric Food Chem,2009,57(3):1051-1059.
[14]贾关荣,赵文恩,申红山,等.西瓜中类胡萝卜素的柱色谱法分离分析[J].安徽农业科学,2008(20):8416-8418.
[15]储大燕.烟草中类胡萝卜素及其异构体的分析研究[D].合肥:中国科学技术大学,2009.
[16]冯作山,陈计峦,孙高峰,等.枸杞色素的提取及纯化技术[J].食品与发酵工业,2004(12):141-144.
[17]彭光华,李忠,张声华.薄层色谱法分离鉴定枸杞子中的类胡萝卜素[J].营养学报,1998(1):79-81.
[18]丁来欣,陈里.薄层色谱法分离测定辣椒红色素中的辣椒红[J].色谱,1995(4):295-296.
[19] Luengo E, Alvarez I, Raso J. Improving carotenoid extraction fromtomato waste by pulsed electric fields[J]. Front Nutr, 2014,1:12.
[20]张玲,郑婷婷,魏伟淋,等.两种不同肉色枇杷果实类胡萝卜素积累及合成相关基因的表达[J].园艺学报,2015(11):2153-2162.
[21]李小丽,梁远学,郜凌超,等.油棕果实不同发育时期类胡萝卜素的含量变化[J].华中农业大学学报,2015(1):23-27.
[22] Richins R D, Kilcrease J, Rodgriguez-Uribe L, et al. CarotenoidExtraction and Quantification from Capsicum annuum[J]. BioProtoc, 2014,4(19).
[23]张海涛,蒋文艳,淡瑞芳.胡萝卜粉中类胡萝卜素的提取工艺优化[J].甘肃农业大学学报,2012(5):139-142.
[24]白婕,韦燕春.金橘类胡萝卜素提取研究[J].大众商务,2009(18):289-290.
[25] Konings E J M, Roomans H H S. Evaluation and validation of anLC method for the analysis of carotenoids in vegetables and fruit[J]. Food Chemistry,1997,59(4):599-603.
[26]胡玉琴,郁继华,王小龙,等.辣椒叶片中类胡萝卜素组分分离及测定方法的研究[J].西北师范大学学报:自然科学版,2015(4):75-80.
[27]王鑫.红辣椒色素的提取、分离及光稳定性研究[D].无锡:江南大学,2005.
[28]戴雄泽,王利群,陈文超,等.辣椒果实发育过程中果色与类胡萝卜素的变化[J].中国农业科学,2009(11):4004-4011.
[29]董彩英.辣椒果实类胡萝卜素的积累特点及其与果色的关系研究[D].扬州:扬州大学,2009.