摘要
酱油和食醋是人们日常生活中重要的调味品。我国于2000年9月1日颁布了酱油和食醋的国家标准,在GB18186-2000《酿造酱油》和GBl8187-2000《酿造食醋》中都规定了要如实标明酱油和食醋生产的方法,即是高盐稀态发酵工艺酱油还是低盐固态发酵发酵工艺酱油,是固态发酵食醋还是液态发酵食醋。但是由于缺乏规范的生产工艺标准和质量检测标准,假冒伪劣产品屡禁不止。
香气成分是评价产品质量的重要指标之一。为了鉴定酱油和食醋的品质,本试验采用顶空固相微萃取-气相色谱法测定酱油和食醋的香气成分。对酱油和食醋的香气成分萃取条件进行了优化及定性、定量研究,并进行了主成分分析和聚类分析,确立了酱油和食醋主要的香气成分和鉴定不同工艺产品的质量指标。结论主要有以下几点:
1.采用固相微萃取-气相色谱法分析酱油中的香气成分,对测定中的萃取头种类、萃取时间、萃取温度、加盐浓度、解析时间等条件参数进行了优化,最终确定了用85μmPA萃取头,在10mL样品中加入1g固体NaCl,萃取温度40℃,萃取时间30min,在气相色谱中解析3min为最佳萃取条件。在此条件下经GC-MS/GC进行定性定量分析,共定性出52种香气组分包括11种酸、6种醛、11种醇、12种酯、4种酚和8种杂环类化合物,定量了其中的20种主要组分。本方法定量重现性良好,标准变异系数(RSD)均小于10%,样品回收率为91.2%~106.7%。
2.采用固相微萃取-气相色谱法分析食醋中的香气成分,对测定中的样品稀释倍数、萃取时间、萃取温度、加盐浓度、解析时间等条件参数进行了优化,最终确定了用85μmPA萃取头,将醋样品用去离子水稀释20倍,取10mL稀释样品中加入3g固体NaCl,萃取温度40℃,萃取时间45min,在气相色谱中解析3min为最佳萃取条件,在此条件下经GC-MS/GC进行定性定量分析,共定性出59种香气组分包括14种醇、12种酯、14种酸、10种醛、3种酚、3种吡嗪和3种杂环类化合物,定量了其中的24种主要组分。本方法定量重现性良好,标准变异系数(RSD)均小于10%,样品回收率为88.3%~100.9%。
3.对酱油的数据进行主成分分析,结果表明:当取5个主成分吋,其累计贡献率达到82.373%>80%,说明前5个主成分能概括原数据的大部分信息。它们包括了酱油中的乙醇、乙酸乙酯、乳酸乙酯、正丁醇、异戊醇、3-甲硫基-1-丙醇、糠醇、β-苯乙醇、4-乙基愈创木酚和2-乙酰基吡咯等重要香气成分。
4.对食醋的数据进行主成分分析,结果表明:当取4个主成分吋,其累计贡献率达到81.253%>80%,说明前4个主成分能概括原数据的大部分信息。它们包括了食醋中的双乙酰、异戊醛、4-乙基愈创木酚、乳酸乙酯、β-苯乙醇、乙酸异戊酯、异丁醇、丁酸、正丁醇和戊酸等重要香气成分。
5.对所测得酱油数据进行聚类分析,发现乙醇、乙酸乙酯、乳酸乙酯、β-苯乙醇和4-乙基愈创木酚在高盐稀态发酵酱油中的含量较高,最高含量分别为33600mg/L、23.3mg/L、36.2mg/L、8.1mg/L和2.8mg/L;而乙酸和2-乙酰基吡咯在低盐固态发酵酱油中的含量较高,分别达到2380.3mg/L和62.9mg/L;且高盐稀态发酵酱油中的香气成分有50多种,在低盐固态发酵酱油中检测到约40种。
经分析还发现:属于高盐稀态发酵工艺的两类酱油在香气成分含量上也存在着很大的差异,除糠醇和2-乙酰基吡咯外,天然晒露发酵酱油中各香气组分的含量比采用日本工艺的高盐稀态发酵酱油的要少。
6.对所测得食醋数据进行聚类分析,发现异戊醛、双乙酰、正丙醇、3-羟基-2-丁酮、乳酸乙酯、糠醛、4-乙基愈创木酚和4-乙基苯酚在固态发酵食醋中的含量较高,最高值分别为3.9mg/L、208.2mg/L、54.9mg/L、2935.7mg/L、19.7mg/L、252.1 mg/L、6.8mg/L和24.3 mg/L;而在液态发食醋中则含量较少,3-羟基-2-丁酮的含量为200.6 mg/L~792.6mg/L,双乙酰的含量为9.7 mg/L~23.3mg/L:除3-羟基-2-丁酮外,以上其它香气成分在配制食醋中则未检出。且固态发酵食醋中的香气成分有50多种,在液态发酵食醋中一般检测到30多种香气成分,而在配制食醋中仅检测到大约20种。
7.综上所述:不同制造工艺的酱油和食醋在香气成分上有明显差异性,这种差异性既体现在物质种类水平上,也表现在相同物质成分的含量水平上,据此差异性可以鉴定不同酿造工艺的酱油和食醋产品。
Soy sauce and vinegar are important seasoning in our daily life.The national standards were promulgated at Sep.1~(st) 2000.The methods for producting soy sauce and viegar must be truthfully regulated in GB 18186-2000 fermented soy sauce and GB 18187-2000 Fermented Vinegar,that was the high-salt liquid-state fermented soy sauce or the low-salt solid-state fermented soy sauces,the solid-state fermented vinegar or the liquid-state fermented vinegar.But there were no standard techniques in soy sauce and vinegar production and the corresponding national quality standards were simple.Therefore,counterfeit products were prevalent.
The flavor component was one of the important indicators to evaluate the quality of some products.Thus,flavor compounds in soy sauce and vinegars were analyzed in the present study by head space-solid phase microextraction coupled with gas chromatogragh(HS-SPME-GC) to assess the quality of products.In this paper, optimization of determination condition for flavor compounds in Soy Sauce and vinegars and were determined qualitatively and quantitatively by means of HS-SPME-GC,and then the data was processed using principal component analysis and cluster analysis,the main flavor components of the soy sauce and vinegars were determined and the quality indexs of different kinds making processes were identified. The main results were as follows.
1.A method for detection of flavor compounds in soy sauce by SPME-GC had been developed.The analysis parameters of SPME fiber coating,extraction time, sample temperature,content of sodium chloride,desorption time had been optimized. Finally,The optimized method was determined:A 10-mL soy sauce was placed into a 20-mL vial and saturated with lg sodium chloride.The sample was extracted using 85μmPA fiber for 30 min at 40℃,then the volatiles were desorbed from the fiber at temperature of 230℃for 3min.Consequently,52 components were identified by means of GC-MS/GC,20 of which were quantified.The identified compounds were 11 alcohols,11 acids,6 carbonyls,12 esters,4 phenols,8 other compounds.The recoveries was between 91.2%and 106.7%.The experiment of reproducibility of relative contents was good and relative standard deviations(RSD) was less than 10%.
2.A method for detection of flavor compounds in vinegars by SPME-GC had been developed.The analysis parameters of dilution multiple,extraction time,sample temperature,content of sodium chloride,desorption time had been optimized.Finally, the optimized method was determined:The vinegar was diluted 20 times with deionized water and then the 10mL diluted vinegar was placed into a 20-mL vial and saturated with 3g sodium chloride.The sample was extracted using 85μmPA fiber for 45 min at 40℃,then the volatiles were desorbed from the fiber at 230℃for 3min. Consequently,59 components were identified by means of GC-MS/GC,24 of which were quantified.The identified compounds were 14 alcohols,14 acids,12 esters,10 carbonyls,3 phenols,3 pyrazines,3 other compounds.The recoveries was at the range from 88.3%to 100.9%.The experiment of reproducibility of relative contents was good and relative standard deviations(RSD) was less than 10%.
3.The results of principal component analysis for the data of the soy sauces indicated that the accumulative contribution rate of the first five principal components to total variation accounted for 82.373%>80%,maintaining most of information of original data.they consisted of alcohol,ethyl acetate,ethyl lactate,butanol,isoamyl alcohol,3-(methylthio)-1-propyl acetate,furfuryl alcohol,β-phenylethyl alcohol, 4-ethyl guaiacol and 2-acetyl pyrrole in soy sauce.
4.The results of principal component analysis for the data of vinegars indicated that the accumulative contribution rate of the first four principal components to total variation accounted for 81.253%>80%,maintaining most of information of original data.they consisted of diacetyl,isovaleraldehyde,4-ethyl guaiacol,ethyl lactate,β-phenylethyl alcohol,isoamyl acetate,isobutanol,butyrate,butanol and pentanoic acid in vinegars.
5.The results of cluster analysis for the data of the soy sauces indicated that the content of alcohol,ethyl acetate,ethyl lactate,butanol,isoamyl alcohol,β-phenylethyl alcohol and 4-ethyl guaiacol were high content in the high-salt liquid-state fermented soy sauce,the maximum content was 33600mg/L,23.3mg/L,36.2mg/L,8.1mg/L and 2.8mg/L respectively;However,the content of acetic acid and 2-acetyl pyrrole was high content in the low-salt solid-state fermented soy sauces,the maximum contents was 2380.3mg/L and 62.9mg/L respectively;There were more than 50 kinds of flavor components detected in the high-salt liquid-state fermented soy sauce,but only approximately 40 kinds of flavor components detected in the low-salt solid-state fermented soy sauces.
The results also showed that there were very great difference for the two kinds of the high-salt liquid-state fermented soy sauce process in the content of the flavor components,except for furfuryl alcohol and 2-acetyl pyrrole.There were lower contents of the flavor compounds in the natural fermented soy sauce than those in the high-salt liquid-state fermented processing adoptd by japanese.
6.The results of cluster analysis for the data of vinegars indicated that the content of isovaleraldehyde,diacetyl,n-propanol,3-hydroxy-2-butanone,ethyl lactate, furfuryl alcohol,4-ethyl guaiacol and 4-ethylphenol was high content in the solid-state fermented vinegar,the maximum contents was 3.9mg/L,208.2rag/L, 54.9mg/L,2935.7mg/L,19.7mg/L,252.1mg/L,6.8mg/L and 24.3 mg/L respectively. However,the content of them were lower content in the liquid-state fermented vinegars.The content of diacetyl was at the range from 9.7mg/L to 23.3 mg/L,the content of 3-hydroxy-2-butanone was at the range from 200.6mg/L to 792.6 mg/L; Except for 3-hydroxy-2-butanone,the other above flavor compounds were not detected in the blended vinegars;There were more than 50 kinds of flavor components detected in the solid-state fermented vinegars,more than 30 kinds of aroma components detected in the liquid-state fermented vinegar,only approximately 20 kinds of flavor components detected in the blended vinegars.
7.The results of principal component analysis and cluster analysis indicated that profiles of flavor components of soy sauce or vinegar depended on their fermentation processes.The difference was reflected in the type and content of flavor compounds. Thus,the fermentation processes of soy sauce or vinegars could be deduced according to the type and content of flavor compounds in their products.
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