氨基酸纸层析展层溶剂初探
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摘要
纸层析法分离鉴定氨基酸试验所用展层溶剂是可以重复使用的,但随着使用次数的增加,展层溶剂中有机溶剂不断挥发,氨基酸的分离效果会越来越差。为了节约试剂、节省试验成本,并改善试验效果,在100 m L重复使用5次的展层溶剂中分别续加0,6,12 m L正丁醇与80%甲酸(体积比5∶1)混合液,混匀,然后与新配制的展层溶剂进行氨基酸纸层析对比试验,观察试验结果,并测定各氨基酸的比移值(Rf)。结果显示,在100 m L重复使用5次的展层溶剂中加入12 m L正丁醇与80%甲酸混合液,其试验结果与用新配制展层溶剂所得试验结果一致。这一结果表明,重复使用多次的层析溶剂可以不废弃,只需添加一定量正丁醇与80%甲酸混合液即可继续使用。这不仅节约了试验成本,而且减少了试验废液的排放,既经济又环保。
The chromatographic solvent which used for separating and identifying amino acids by paper chromatography, could be reused. However, with the usage count of chromatographic solvent increase, the organic solvent was continuously volatilized, and the separation effect of the amino acid was getting worse. To save reagents, economize experiment costs, and improve experimental results, 0,6, 12 m L of n-butanol and 80% formic acid(5∶1 by volume)were added, respectively, into 100 m L chromatographic solvent which had used for 5 times. Then, mixed the above reagent, we designed a comparative experiment with the used and reconfigured chromatographic solvent. The experimental results were observed, and the Rf value of each amino acid was determined. The result showed that added 12 m L of a mixture of n-butanol and 80% formic acid into 100 m L chromatographic solution used repeatedly for 5 times, was consistent with that using the newly prepared chromatographic solvent. These results indicated that the chromatographic solution that was repeatedly used could not be discarded, simply by adding a certain amount of n-butanol and 80% formic acid mixture, it could be used continuously. This would not only help to save the experimental funds, but also reduce the discharge of experimental waste liquid. This way was both economical and environmentally friendly.
The chromatographic solvent which used for separating and identifying amino acids by paper chromatography, could be reused. However, with the usage count of chromatographic solvent increase, the organic solvent was continuously volatilized, and the separation effect of the amino acid was getting worse. To save reagents, economize experiment costs, and improve experimental results, 0,6, 12 m L of n-butanol and 80% formic acid(5∶1 by volume)were added, respectively, into 100 m L chromatographic solvent which had used for 5 times. Then, mixed the above reagent, we designed a comparative experiment with the used and reconfigured chromatographic solvent. The experimental results were observed, and the Rf value of each amino acid was determined. The result showed that added 12 m L of a mixture of n-butanol and 80% formic acid into 100 m L chromatographic solution used repeatedly for 5 times, was consistent with that using the newly prepared chromatographic solvent. These results indicated that the chromatographic solution that was repeatedly used could not be discarded, simply by adding a certain amount of n-butanol and 80% formic acid mixture, it could be used continuously. This would not only help to save the experimental funds, but also reduce the discharge of experimental waste liquid. This way was both economical and environmentally friendly.
引文
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[5]张敏,薛正莲,余飞,等.纸层析-酶标仪法测定发酵液中γ-氨基丁酸[J].食品与发酵工业,2019,40(7):156-160.
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[13]王新洲,郝旭晨.一种常温常压下有机溶剂脱色除臭脱酸干燥处理装置:CN204485848U[P].2015-07-22.
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[16]王志成,姚安娜,田博,等.氨基酸纸层析实验方法改进及未知红色条带的讨论[J].科协论坛,2008(2):70-71.
[17]田明,高铮,苏英华,等.氨基酸纸层析实验的进一步探讨[J].实验室科学,2009,28(6):64-65.
[18]杨晓玲,赵宗云,刘永军,等.应用纸层析分离鉴定氨基酸方法的改进[J].植物生理学报,2002,38(4):368.
[19]陈倩,刘晶,张璐.纸层析法分离鉴定氨基酸实验的改进[J].化工管理,2017(12):25-26.
[20]王海鸥,周丽亚.纸层析法分离鉴定氨基酸实验的改进[J].广州化工,2017,45(6):117-118.
[21]刘颖沙,刘小宁,李国秀,等.氨基酸纸色谱层析法实验条件优化的探索[J].杨凌职业技术学院学报,2019,18(1):21-24.
[2]杨丽,尤丽,叶金秀,等.纸层析分离鉴定氨基酸实验的改进[J].云南民族大学学报(自然科学版),2011,20(3):229-231.
[3]叶芳,胡燕,宋玉梅.纸层析法分离氨基酸相关研究进展[J].科技展望,2017,27(13):83-91.
[4]朱彩瑜,赵雨彤,刘先俊,等.关于氨基酸分离的创新层析实验的探究[J].教育教学论坛,2019(1):274-278.
[5]张敏,薛正莲,余飞,等.纸层析-酶标仪法测定发酵液中γ-氨基丁酸[J].食品与发酵工业,2019,40(7):156-160.
[6]THRKOW B,WEUFFEN W.Gas chromatographic determination of thiocyanate in vegetable and animal foods and in human serum following paper chromatographic separation[J].Die Pharmazie,1981,36(2):96-99.
[7]刘维全.动物生物化学实验指导[M].北京:中国农业大学出版社,2014:41-43.
[8]胡兰.动物生物化学实验教程[M].北京:中国农业大学出版社,2006:32-37.
[9]张华芳.L-丝氨酸分离提取的研究进展[J].化工设计通讯,2018(9):196,250.
[10]李海霞,王晶,何莹,等.反相纸层析法分离油菜籽中脂肪酸的经验总结[J].生物学杂志,2019(2):121-123.
[11]郑亚玲.复方多糖的葡聚糖凝胶分离及纯度鉴定[J].食品安全导刊,2018(22):58-61.
[12]李文,董明盛.发酵鹰嘴豆乳产γ-氨基丁酸乳酸菌的复合诱变选育[J].食品科学,2018(1):147-153.
[13]王新洲,郝旭晨.一种常温常压下有机溶剂脱色除臭脱酸干燥处理装置:CN204485848U[P].2015-07-22.
[14]杨黎华.我国挥发性有机溶剂滥用的预防[J].法制与社会,2011(33):211-212.
[15]陈舒丽,王梅娟,罗丽丹,等.对氨基酸纸层析实验的改进[J].山西医科大学学报,2011,42(3):273-274.
[16]王志成,姚安娜,田博,等.氨基酸纸层析实验方法改进及未知红色条带的讨论[J].科协论坛,2008(2):70-71.
[17]田明,高铮,苏英华,等.氨基酸纸层析实验的进一步探讨[J].实验室科学,2009,28(6):64-65.
[18]杨晓玲,赵宗云,刘永军,等.应用纸层析分离鉴定氨基酸方法的改进[J].植物生理学报,2002,38(4):368.
[19]陈倩,刘晶,张璐.纸层析法分离鉴定氨基酸实验的改进[J].化工管理,2017(12):25-26.
[20]王海鸥,周丽亚.纸层析法分离鉴定氨基酸实验的改进[J].广州化工,2017,45(6):117-118.
[21]刘颖沙,刘小宁,李国秀,等.氨基酸纸色谱层析法实验条件优化的探索[J].杨凌职业技术学院学报,2019,18(1):21-24.