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双孢蘑菇汤特征风味物质的鉴定及熬制过程风味物质释放规律研究
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摘要
双孢蘑菇是我国培养最广、产量最大的一种食用菌。目前对食用菌的利用主要停留于干制、腌渍及罐装等粗加工形式,其精深加工力度不足,严重阻碍了食用菌产业的发展。近年来,国内外研究者对双孢蘑菇的研究主要集中在营养成分、功能性质及风味成分的研究方面,对其深加工产品的风味研究鲜见报道。双孢蘑菇因其营养丰富、风味独特成为制作汤品的重要原料,研究蘑菇汤的特征风味物质及熬制过程风味物质的释放规律,对双孢蘑菇精深加工具有重要的理论指导意义和实用价值。
     本论文以双孢蘑菇为研究对象,对双孢蘑菇汤的特征滋味物质和特征挥发性风味物质进行鉴定,比较不同烹调方法对双孢蘑菇汤风味的影响,研究熬制过程风味的释放规律,采用外源加入法初步探讨风味的形成机理,并采用酶解技术促进风味物质的释放。主要研究内容如下:
     结合仪器分析(HPLC)与感官分析对双孢蘑菇汤的特征滋味物质进行分析,利用滋味活度值(TAV)确定双孢蘑菇汤的特征滋味物质,结果表明,甘露醇、谷氨酸及Ca2+是双孢蘑菇汤的特征滋味物质。但由于核苷酸类与鲜味氨基酸之间存在协同增效作用,因此,鲜味核苷酸对双孢蘑菇汤的贡献也不容忽视。
     采用同时蒸馏萃取(SDE)和顶空固相微萃取(HS-SPME)两种萃取方法对双孢蘑菇汤的挥发性风味物质进行提取,通过气质联用技术(GC/MS)对其组成及含量进行分析,结果发现,含8碳原子的不饱和醇、醛、酮类、芳香醛类及杂原子类化合物是双孢蘑菇汤的主要挥发性风味物质。两种萃取方式得到的双孢蘑菇汤的挥发性风味物质差异较大,SPME法萃取得到50种挥发性风味物质而SDE法得到41种挥发性风味物质且SPME得到8碳原子的不饱和醇、醛、酮类、芳香醛类及杂原子类化合物的相对含量高于SDE。因此,SPME更适合于双孢蘑菇汤挥发性风味物质的提取。对影响SPME萃取效果的主要因素包括萃取头、萃取时间和萃取温度进行了选择,得到萃取头DVB/CAR/PDMS比较适合于双孢蘑菇汤风味物质的萃取,最佳萃取条件为55℃、30 min。同时比较了弱极性的HP-5与极性的DB-WAX两种色谱柱对双孢蘑菇汤风味物质的影响,结果显示DB-WAX色谱柱更适合于双孢蘑菇汤风味物质的分析。
     在以上最佳萃取条件下,进一步利用气相色谱-嗅闻技术(GC/O)对SPME法提取得到的双孢蘑菇汤的风味物质进行分析。分别采用香味萃取物稀释分析(AEDA)和时间强度法(OSME)两种GC/O定量分析方法分析其中的特征挥发性风味物质,结果显示双孢蘑菇汤的主要特征挥发性风味物质包括蘑菇味的1-辛烯-3-酮、1-辛烯-3-醇、熟蘑菇味的3-辛烯-2-酮、烤坚果味的2, 6-二甲基吡嗪、花香味的苯乙醛、果香味6-甲基-5-庚烯-2-酮、甜香味的二氢-5-戊基-2(3H)-呋喃酮、清香味的正己醇、巧克力味的2-乙烯基-6-甲基吡嗪、肉香味的2-乙酰噻唑及未知的呈爆米花味、酱香味、果香味的三种化合物。
     研究了传统熬制、高压及微波加工三种不同的烹调方式对双孢蘑菇汤风味的影响,分别利用仪器分析(HPLC、GC/MS、GC/MS/O、电子鼻)及感官评定分析了不同的烹调方法对双孢蘑菇汤的滋味物质、挥发性风味物质及整体风味特征的影响。仪器分析结果显示因各种烹调方法的加热方式不同,所以双孢蘑菇汤的风味物质组成存在显著差异,其中等鲜浓度(EUC)在微波熬制的汤中最高,其次为传统熬制的汤;感官分析结果显示传统熬制的汤的鲜味强度及风味的整体接受性得分最高。因此,传统熬制的双孢蘑菇汤风味最佳。
     为了对双孢蘑菇汤的风味品质进行控制,进一步研究了双孢蘑菇汤在传统熬制过程中风味物质及感官风味特征的变化,并分析了风味物质与感官风味特征的相关性。结果表明,非挥发性呈味物质含量在熬制起始阶段显著增加,在30~60 min间达到一个动态平衡,之后含量有所降低;双孢蘑菇汤的多种特征风味物质在熬制前30 min都呈增加趋势,而60 min后有些物质显著降低或消失。因此,熬制时间过长对双孢蘑菇汤风味不利。排序法感官分析结果显示,双孢蘑菇汤的鲜味强度与EUC相关,但总体风味可接受性得分与EUC相关性不大,这说明蘑菇汤风味不仅受鲜味物质的影响,同时受其它风味物质的影响。为了进一步了解各种风味物质对感官风味特征的贡献,利用偏最小二乘回归(PLSR)分析了特征风味物质与感官描述符的相关性,可以看出,特定的感官属性受特定的风味物质的影响。
     通过外源性的加入风味前体物质——氨基酸,对双孢蘑菇汤熬制过程风味形成的机理进行初步探讨,结果显示Ala、Leu及Arg的加入使醛类总量显著增加,Leu的加入使3-甲基丁醛的含量显著增加,Met的加入使含硫化合物如3-甲硫基丙醛、2-乙酰噻唑;氨基酸的加入对醇类物质的含量影响不显著;而杂原子化合物的量显著增加,其中加入丙氨酸和亮氨酸的汤中吡嗪类化合物显著增加。研究了不同种类的酶解预处理对双孢蘑菇汤的营养成分与风味成分的影响。结果表明:经过酶解预处理,双孢蘑菇汤中的固形物、蛋白质、游离氨基酸等物质的含量都显著提高;食用菌水解酶的作用效果比纤维素酶和其他蛋白酶双酶分步酶解的效果显著。
Button mushroom (Agaricus bisporus (Lange) Sing) is the most popular cultivated mushroom in China. It is currently used to produce fresh mushrooms, dried mushrooms, canned mushrooms and other rough forms, but the deep processing of this edible mushroom is insufficient. This hinders the development of the mushroom industry. Nowadays, the nutrition and functional properties of button mushroom were studied, but there were few studies about its flavour compounds and products. Button mushroom was a good material for producing soup due to its rich nutrients and unique flavour. The research on the characteristic flavour active compounds of mushroom soup and flavour release during cooking process has important theoretical significance and practical value for deep processing of button mushroom industry.
     Button mushroom was used to produce mushroom soup in this study. The characteristic aroma-active compounds and taste-active compounds were identified by instrumental analysis and sensory analysis. Effect of different cooking methods on the flavour constituents of mushroom (Agaricus bisporus L.) soup was studied. Flavour releasing of mushroom soup during cooking was investigated. The mechanism of the flavour formation was explored by adding some precursors. Enzyme technology was used for releasing the nutrition and flavour of mushroom. The main content of this thesis was as follows:
     Instrumental analysis (HPLC) and sensory evaluation were used to analyze the characteristic taste compounds of mushroom soup. Taste activity value (TAV) was used to determine the characteristic taste compounds in the soup. The result showed that mannitol, Glu and Ca2+ were the characteristic taste compounds. Because of the synergistic interaction between umami amino acids and 5'- nucleotides, the 5'- nucleotides was regarded as the characteristic taste compounds.
     SDE and HS-SPME were used to extract the volatile compounds and GC/MS was used to identify constitute and content of volatile compounds. The identified aroma compounds included C8 derivatives of octane and octene, such as 1-octen-3-ol, trans-2-octen-1-ol, 1-octen-3-one, benzaldehyde derivatives and heterocyclic compounds. SPME was proved to detect more flavour compounds (50) than SDE (41), and the relative contents of C8 derivatives of octane and octene, benzaldehyde derivatives and heterocyclic compounds extracted by SPME were higher than those extracted by SDE. Therefore, SPME was chosen as the extraction method. The effect of extraction conditions ( fiber, extraction time and temperature) on the volatiles in mushroom soup was studied, and the result indicated that DVB/CAR/PDMS fiber was suitable for mushroom soup flavour extraction and the flavour was extracted at 55°C for 30 min. HP-5 and DB-WAX columns were used to separate the volatiles, and DB-WAX column was suitable for the separation of volatile compounds.
     GC/O was used to determine the aroma-active compounds by Aroma Extract Dilution Analysis (AEDA) and time-intensity method (OSME) analysis. Mushroom flavour (1-octen-3-one), cooked mushroom flavour (3-octen-2-one), roast nut flavor (2, 6-dimethylpyrazine), green flavor (1-hexanol) and fruity flavor (dihydro-5-pentyl-2(3H)-furanone), popcorn flavor (unknown), sauce flavour (unknown), floral (benzeneacetaldehyde) are the main flavour of the mushroom soup.
     Effect of cooking methods on the flavour constitutes and flavour profile was analysed by instrumental analysis (HPLC, GC/MS, GC/MS/O, Electronic nose) combined with sensory evaluation. Results showed that the flavour constitutes were significantly different depending on different heating treatment. The umami intensity was highest in traditional cooking mushroom soup, but the EUC was highest in microwaved mushroom soup. It indicated that umami amino acids and nucleotides were one of the factors affecting the umami taste.
     The flavour constitutes and sensory evaluation changes of mushroom soup during cooking process were determined by instrumental and sensory analysis. The nonvolatile compounds were increased before 30 min of cooking and up to a dynamic equilibrium between 30-60 min, then decreased after 60 min. It indicated that the nonvolatile compounds not only contributed to the taste of mushroom soup, but were the precursors of volatile compounds. Most of the aroma-active compounds increased during the first 30 min cooking time and decreased or disappeared after 60 min of cooking. Therefore, long time cooking was negative for the mushroom soup flavour. The rank-order test result showed that the umami intensity was positively correlated to EUC, but the total acceptability score was not related to the EUC. It indicated that mushroom soup flavour was due to the aroma-active compounds. The correlation between the aroma-active and umami-related compounds’concentration and the sensory attributes was analysed by partial least square regression (PLSR). Correlations between the flavour compounds and sensory attributes of mushroom soup indicated that specific flavour active compounds could be associated with specific sensory attributes.
     The mechanism of volatile flavour formation in mushroom soup during cooking process was preliminaryly explored by adding amino acids. The result indicated that the impact of amino acids addition on aldehydes, ketones and heterocyclic compounds formation was significant. 3-methyl butanal increased with Leu addition, and methional, 2-acetylthiazole increased with Met addition. Pyrazine compounds increased with Ala and Leu addition.
     The effect of enzymatic pre-treatment on the nutrition and flavour of mushroom soup was investigated. The results indicated that the levels of solid, protein and free amino acids increased significantly in the mushroom soup compared with the control, and the effect of edible fungus hydrolase was better than cellulase and other protease.
引文
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