为了探究茚三酮比色法在黄酒和米酒氨基酸态氮测定中的适用性,首先对可能影响茚三酮比色法测定结果的黄酒和米酒特性因素进行了考察分析,然后与甲醛滴定法及氨基酸分析仪进行比较,对17个黄酒和米酒样品进行了测定分析。结果表明,黄酒和米酒的色泽、总酸和酒精度等特性,对茚三酮比色法的测定结果无显著影响。茚三酮比色法与甲醛滴定法测定结果之间具有较好的线性关系,但对于米酒等氨基酸态氮含量较低的样品,茚三酮比色法具有更高的精密度和准确性。对照氨基酸分析仪分析结果,甲醛滴定法更适于表征产品的总体氨基酸含量,而茚三酮比色法则能更准确反映发酵过程中的α-氨基酸含量,且对于氨基酸态氮含量较低的米酒产品测定结果相对更为准确。
The applicability of ninhydrin colorimetric method for amino-acid nitrogen (AAN) measurement from Huangjiu and Mijiu analysis was determined in this work. First, factors of Huangjiu and Mijiu that may affect ninhydrin colorimetry result were investigated. Next, ninhydrin colorimetry, formaldehyde titration, and the amino acid analyzer were used for AAN measurement of 17 Huangjiu and Mijiu samples. The results showed that color, total acid, and alcohol content of Huangjiu and Mijiu had no significant effect on the ninhydrin colorimetry determination. There was a good linear relationship between the results given by ninhydrin colorimetry and formaldehyde titration. However, the ninhydrin colorimetry had higher precision and accuracy for the samples with low amino acid nitrogen content. Compared to the result of the amino acid analyzer, the formaldehyde titration was more suitable to determine the total amino acid content. Moreover, the ninhydrin colorimetry was able to accurately determine the α-amino acid content during the fermentation process. These results not only provide an alternative analysis method for amino acid nitrogen in Huangjiu, but also establish basis for the developing amino acid nitrogen analysis index in Mijiu industry.
[1] YANG Y, XIA Y, WANG G, et al.Effects of boiling, ultra-high temperature and high hydrostatic pressure on free amino acids, flavor characteristics and sensory profiles in Chinese rice wine[J]. Food Chemistry, 2019, 275: 407-416.
[2] YU H Y, ZHAO J, LI F H, et al.Characterization of Chinese rice wine taste attributes using liquid chromatographic analysis, sensory evaluation, and an electronic tongue[J]. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2015, 997: 129-135.
[3] 尹花,贺扬,董建军,等.麦汁关键氨基酸对Lager酵母发酵性能的影响[J].食品与生物技术学报, 2018, 37(9): 987-993.
[4] 潘慧青,曹钰,石慧媛,等.黄酒酿造后酵工艺对氨基酸态氮生成的影响[J].食品与生物技术学报, 2016, 35(2): 144-150.
[5] GB/T 13662—2018,黄酒[S].北京:中国国家标准化管理委员会, 2018: 28.
[6] GB 5009.124—2016,食品安全国家标准,食品中氨基酸的测定[S].北京:国家食品药品监督管理总局,2016: 12.
[7] SQUIRE R A,CAMERON L L.An analysis of potential carcinogenic risk from formaldehyde[J].Regulatory Toxicology and Pharmacology, 1984, 4(2):107-129.
[8] 崔敏,傅婕,迟原龙,等.茚三酮比色法和甲醛滴定法测定水解胶原的比较[J].中国皮革,2011,40(7):1-4.
[9] 管敦仪.啤酒工业手册(中册)[M].北京:中国轻工业出版社,1982:191-193.
[10] ABERNATHY D G, SPEDDING G, STARCHER B. Analysis of protein and total usable nitrogen in beer and wine using a microwell ninhydrin assay[J]. Journal of the Institute of Brewing, 2009, 115(2): 122-127.
[11] 涂云飞.茚三酮法测定茶叶游离氨基酸总量研究[J].现代农业科技,2018(14):235;238.
[12] HE X, WU Z, YE Y, et al. Research progress on detection of amino acids by high performance liquid chromatography[J]. Journal of Instrumental Analysis, 2016, 35(7): 922-928.
[13] 迟原龙,文振华,崔敏,等.胶原水解度的测定[J].中国皮革,2010, 39(19):13-16.
[14] HERBERT P, BARROS P, RATOLA N, et al. HPLC determination of amino acids in musts and port wine using OPA/FMOC derivatives[J]. Journal of Food Science, 2000, 65(7):1 130-1 133.
[15] 张永国,秦锐兰.麦汁中α-氨基氮含量的测定方法[J].啤酒科技,2008(3):35-37.
[16] 惠明,邱聪,张会娟,等.老户周黄酒色泽与易褐变营养物含量的相关性研究[J].河南工业大学学报(自然科学版),2018,39(2):15-19.
[17] 岳元媛,田艳.鲜味提取物中氨基酸态氮测定方法的比较研究[J].中国井矿盐,2015,46(1):39-43.
[18] MENDEL F. Applications of the ninhydrin reaction for analysis of amino acids, peptides, and proteins to agricultural and biomedical sciences[J]. Journal of Agricultural & Food Chemistry,2004,52(3):385-406.
[19] 李绍军,龚月桦,王俊儒,等.关于茚三酮法测定脯氨酸含量中脯氨酸与茚三酮反应之探讨[J]. 植物生理学通讯,2005(3):365-368.
[20] TUR’YAN. Theoretical bases of the ammonium ion determination by formol titration[J]. Reviews in Analytical Chemistry, 2010, 29(1):25-37.
[21] LIE S. The EBC-Ninhydrin method for determination of free alpha amino nitrogen[J]. Journal of the Institute of Brewing,1973(79):37-41.
[22] 顾国贤.酿造酒工艺学[M].北京:中国轻工业出版社.2009:456-457.
