研究论文

饲料中添加高水平蚕豆对吉富罗非鱼肌肉中挥发性风味物质组成的影响

  • 杨经群 , 1, 2 ,
  • 彭凯 1, * ,
  • 黄文 1 ,
  • 胡俊茹 , 1, ** ,
  • 赵红霞 1 ,
  • 曹俊明 1, 2, 3 ,
  • 陈冰 , 1, **
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  • 1 广东省农业科学院动物科学研究所,广东省农业科学院水产研究中心,农业农村部华南动物营养与饲料重点实验室,广州 510640
  • 2 广东海洋大学水产学院,湛江 524088
  • 3 广东省农业科学院,广州 510640
** 胡俊茹,副研究员,硕士生导师,E-mail: ;
陈 冰,副研究员,硕士生导师,E-mail:

*同等贡献作者

杨经群(1999—),女,广西梧州人,硕士研究生,从事水产动物营养与饲料研究。E-mail:

Office editor: 菅景颖

收稿日期: 2025-01-10

  网络出版日期: 2025-09-12

基金资助

中央财政农业发展(优势特色产业集群)资金“脆肉罗非鱼专用饲料的研制及试验”(2022横001)

茂名市科技计划项目(2022DZXHT066)

Effects of Diet with High Level of Faba Bean on Muscle Volatile Flavor Substances Composition of Genetic Improvement of Farmed Tilapia (Oreochromis niloticus)

  • YANG Jingqun , 1, 2 ,
  • PENG Kai 1 ,
  • HUANG Wen 1 ,
  • HU Junru , 1, ** ,
  • ZHAO Hongxia 1 ,
  • CAO Junming 1, 2, 3 ,
  • CHEN Bing , 1, **
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  • 1 Key Laboratory of Animal Nutrition and Feed in South China, Ministry of Agriculture and Rural Affairs, Fisheries Research Center of Guangdong Academy of Agricultural Sciences, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
  • 2 College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China
  • 3 Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
** HU Junru, associate professor, E-mail: ;
CHEN Bing, associate professor, E-mail:

*Contributed equally

Received date: 2025-01-10

  Online published: 2025-09-12

摘要

为了探讨在吉富罗非鱼饲料中添加高水平蚕豆对其肌肉中挥发性风味物质组成的影响,本试验选取初始体质量为(402.32±8.59) g的吉富罗非鱼50尾,随机分成2组(每组5个重复,每个重复5尾鱼),在49 d的饲养期中分别投喂含0(对照组,记为G0组)和70%(试验组,记为G70组)蚕豆的等氮等脂试验饲料。采用全二维气相色谱-飞行时间质谱(GC×GC-TOF MS)技术对2组罗非鱼肌肉样本进行风味组学分析。结果显示:1)在2组罗非鱼肌肉样本中共检测到920种挥发性风味物质,其中G0组检测出707种,G70组检测出633种,2组都检测到的挥发性风味物质有420种。2)罗非鱼肌肉中的挥发性风味物质种类主要包括醇类、醛类、羧酸类、酯类、杂环化合物、烃类、酮类等7类,其中,G0组醇类的相对含量最高(32.97%),其次是酮类(10.82%);G70组也是醇类的相对含量最高(40.52%),其次是酯类(13.98%)。3)2组罗非鱼肌肉中呈显著性差异的挥发性风味物质有10种,与G0组相比,G70组6-甲基-5-庚烯-2-酮、5-甲基-2-己酮、苯乙酸乙酯、1-辛烯-3-醇、1-甲基吡咯、(Z)-6-十一烯-2-酮、2-羟基-3-戊酮的含量显著降低(P<0.05),6-甲基-2,4-庚二酮、异戊酸乙酯、1-巯基-2-丙酮的含量显著提高(P<0.05)。4)罗非鱼肌肉中的关键挥发性风味物质为2,3-丁二酮、2-甲基丁醛、2-十一烷、2-戊基呋喃。与G0组相比,G70组2,3-丁二酮(贡献黄油香味)的相对气味活度值(ROVA)降低,2-甲基丁醛(贡献可可香味)的ROVA提高。综合上述结果得出,在本试验条件下,饲料中添加高水平(70%)蚕豆提高了吉富罗非鱼肌肉中醇类和酯类的相对含量,提高了2-甲基丁醛的ROVA以及降低了2,3-丁二酮的ROVA,使吉富罗非鱼肌肉脂肪风味发生变化;此外,饲料中添加高水平蚕豆略微降低了吉富罗非鱼整体肌肉的水果风味,并可抑制腥味物质的作用,降低肌肉中的土腥味。

本文引用格式

杨经群 , 彭凯 , 黄文 , 胡俊茹 , 赵红霞 , 曹俊明 , 陈冰 . 饲料中添加高水平蚕豆对吉富罗非鱼肌肉中挥发性风味物质组成的影响[J]. 动物营养学报, 2025 , 37(9) : 6153 -6162 . DOI: 10.12418/CJAN2025.500

Abstract

In this experiment, to explore the effects of diet with high level of faba bean on the muscle volatile flavor substances composition of genetic improvement of farmed tilapia (GIFT, Oreochromis niloticus), fifty GIFT with initial body weight of (402.32±8.59) g were randomly divided into 2 groups, with 5 replicates per group and 5 fish per replicate. During the feeding period of 49 days, the two groups of experimental fish were fed with isonitrogenous and isolipidic diets containing 0 (control group, denoted as G0 group) and 70% (experimental group, denoted as G70 group) faba bean, respectively. The two groups of tilapia muscle samples were sent for the determination of flavoromics using comprehensivetwo-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOF MS). The results showed as follows: 1) a total of 920 volatile flavor substances were detected in tilapia muscle samples in two groups, 707 in G0 group, 633 in G70 group, and a total of 420 volatile flavor substances were detected in both groups. 2) The volatile flavor substances in the tilapia muscle mainly included 7 categories, including alcohols, aldehydes, carboxylic acids, esters, heterocyclic compounds, hydrocarbons and ketones, among which G0 group had the highest relative content of alcohols (32.97%), followed by ketones (10.82%), while G70 group had the highest relative content of alcohols (40.52%), followed by esters (13.98%). 3) It was found that there were 10 kinds of volatile flavor substances in tilapia muscle with significant differences between the two groups. Compared with group G0, the contents of 6-methyl-5-heptene-2-ketone (C11H20O), 5-methyl-2-hexanone (C7H14O), (Z)-undecen-2-one (C11H20O), ethyl phenylacetate (C10H12O2), 1-octen-3-ol (C8H16O), 1-methylpyrrole (C5H7N), 2-hydroxy-3-pentanone (C5H10O2) in the group G70 were significantly decreased (P<0.05), while the contents of 6-methyl-2,4-heptanedione (C8H14O2), ethyl isovalerate (C7H14O2) and 1-mercapto-2-propanone (C3H6OS) were significantly increased (P<0.05). The key volatile flavor substances of tilapia muscle are 2,3-butanedione (C4H6O2), 2-methylbutyraldehyde (C5H10O),2-heneicosane (C11H24) and 2-pentylfuran (C9H14O). Compared with G0 group, the relative odor activity value (ROVA) of C4H6O2 (contributing to butter flavor) in G70 group was decreased, while the ROVA of C5H10O (contributing to cocoa flavor) was increased. To sum up, under the experimental conditions, adding high level (70%) faba bean to the diet can improve the relative contents of alcohols and esters in the muscle of GIFT, increase the ROVA of C5H10O and decrease the ROVA of C4H6O2, which can change the fat flavor of tilapia muscle. Besides, diets supplemented with high level of faba bean slightly reduces the fruit flavor of muscle of GIFT, and inhibits the action of fishy substances, reducing the earthy smell of muscle.

吉富罗非鱼具有生长速度快、耐低氧、繁殖能力强、产量高等特点,且背宽肉厚、出肉率高、无肌间刺,富含人体所需的8种氨基酸,营养价值高,是我国南方养殖的主要经济鱼种之一。2023年,我国罗非鱼总产量超181万t[1]。肌肉硬度(紧实度)是高品质鱼类的重要评价指标之一,也是影响鱼片受喜爱程度的重要因素[2]。基于提高罗非鱼养殖利润和鱼肉口感的目的,市场上出现了用蚕豆投喂的罗非鱼,不同于普通罗非鱼,其肉质具有更高的硬度和咀嚼性[3-5]
随着人们生活水平的提高,除了质量外,肉品的风味和口感也越来越受到消费者的重视。鱼肉的风味和口感与一系列代谢产物密切相关,根据溶解度的不同,可将其分为两大类,即水溶性和脂溶性。水溶性前体化学品主要由维生素、氨基酸和核苷酸组成。其中,游离氨基酸,如谷氨酸和丝氨酸,在很大程度上决定了肉的风味和味道[6],而肌苷5'-单磷酸影响肉的鲜味和新鲜度。脂溶性前体化学品大多是脂类代谢物,其中脂肪酸在氧化时可转化为挥发性呈味化合物,如醛、酮和醇,这些化合物有助于增强肉的风味[7]。这些代谢产物可以通过协同作用和相互作用影响肉类的整体风味和口感。迄今为止,尽管饲料中添加蚕豆对改善鱼肉品质影响的研究已有许多报道,但致力于蚕豆对鱼肌肉的风味和口感影响的研究还相对较少。因此,本试验以吉富罗非鱼为试验对象,研究饲料中添加高水平(70%)的蚕豆对其肌肉中挥发性风味物质组成的影响,以期为改善鱼肉风味提供参考。

1 材料与方法

1.1 试验饲料

首先配制以豆粕、菜籽粕为主要蛋白质源且未添加蚕豆的饲料(记为G0),然后用蚕豆替代G0中部分原料使蚕豆的占比达到70%,同时添加鱼粉调节其粗蛋白质含量,补充适量的晶体赖氨酸(纯度为99%)和晶体蛋氨酸(纯度为99%)以补足赖氨酸、蛋氨酸,并通过微量调节油脂的配比,配制成与G0等氮等脂的饲料(记为G70)。试验饲料组成及营养水平见表1。各饲料原料粉碎后均经过60目筛,按照配比混合后搅拌均匀,用T52水产饲料膨化机(广东华强膨化机械厂)制成膨化料(110 ℃膨化)后,喷油、烘干(55 ℃),4 ℃储存备用。
表1 试验饲料组成及营养水平(干物质基础)

Table 1 Composition and nutrient levels of experimental diets (DM basis) %

项目
Items
饲料Diets
G0 G70
原料Ingredients
鱼粉Fish meal 3.00 8.90
豆粕Soybean meal 30.00 8.00
菜籽粕Rapeseed meal 30.00
磷脂Phosphatide 2.00 2.00
豆油Soybean oil 2.00 1.10
面粉Flour 5.00
α-淀粉α-starch 15.00
磷酸二氢钙Ca(H2PO4)2 1.50 1.50
矿物质预混料Mineral premix1) 1.00 1.00
维生素预混料Vitamin premix2) 0.20 0.20
微晶纤维素
Microcrystalline cellulose
9.64 6.94
蚕豆粉Faba bean meal 70.00
晶体蛋氨酸Crystal methionine 0.27 0.30
晶体赖氨酸Crystal lysine 0.39 0.06
合计Total 100.00 100.00
营养水平Nutrient levels3)
粗蛋白质Crude protein 27.85 27.86
粗脂肪Crude lipid 5.03 5.04
蛋氨酸Methionine 0.54 0.54
赖氨酸Lysine 2.08 2.09

1)每千克矿物质预混料含有 One kilogram of mineral premix contained the following:MgSO4·H2O 12 g,Ca(IO3)2 230 g,Met-Cu 1.5 g,Met-Co 0.25 g,KCl 36 g,FeSO4·H2O 1 g,Na2SeO3 0.05 g,ZnSO4·H2O 10 g,MnSO4·H2O 1.9 g。

2)每千克维生素预混料含有 One kilogram of vitamin premix contained the following:叶酸 folic acid 1.28 g,泛酸钙 calcium pantothenate 16 g,烟酸 nicotinic acid 28 g,肌醇 inositol 40 g,生物素 biotin 64 mg,VB12 16 mg,VB1 4 g,VK 4 g,VB6 4.8 g,VB2 8 g,VE 16 g,VA 3 200 000 IU,VD3 1 600 000 IU。

3)粗蛋白质(GB/T 6432—2018)、粗脂肪(GB/T 6433—2006)和氨基酸(GB/T 18246—2019)均为参考国标方法所得实测值。Crude protein (GB/T 6432—2018), crude lipid (GB/T 6433—2006) and amino acids (GB/T 18246—2019) were measured values according to the national standard methods。

1.2 饲养试验

动物试验已经过广东省农业科学院动物科学研究所审查伦理和批准(批准编号2023009)。将初始体重为(402.32±8.59) g的50尾吉富罗非鱼随机分成2组,每组5个重复,每个重复5尾。2组分别投喂未添加蚕豆的饲料(对照组,记为G0组)和添加70%蚕豆的饲料(试验组,记为G70组),在广东农业科学院动物科学研究所的室内温控循环养殖系统中进行为期49 d的饲养试验。
每天投喂2次(09:00和17:00),每次投喂量约鱼体重的2%,每天记录饲料投喂量、水质情况以及试验鱼死亡情况。养殖期间,24 h供氧,水体中溶氧含量>7 mg/L,氨氮含量<0.05 mg/L,亚硝酸盐含量<0.01 mg/L,水温在29.5~31.5 ℃。

1.3 样品采集

饲养试验结束后进行样品采集。每个重复随机取1尾罗非鱼,采样前禁食24 h,经过放血刮鳞去皮处理后,取背部肌肉,切成质量均匀的肉块(3.0±0.1) g,用生理盐水冲洗掉表面污渍后装入冻存管,液氮速冻后放入-80 ℃冰箱保存待测。

1.4 肌肉中挥发性风味物质的提取与测定

取适量罗非鱼肌肉样本于15 mL离心管中,将离心管放入泡沫箱中并加入适量液氮冷冻5 min;将样本转移至打碎机中,打碎3 min;取1 g样本于20 mL顶空进样瓶中,加入10 μL内标溶液于样品中;将转移后的样本在60 ℃条件下孵育30 min;吸附样本前,SPME萃取头在270 ℃条件下老化10 min;将老化后的SPME萃取头转移至孵育室,在60 ℃条件下吸附样本40 min;吸附结束后,将SPME萃取头转移至GC进样口,在250 ℃条件下脱附5 min;进样后,SPME萃取头在270 ℃条件下老化10 min。取10 μL正构烷烃至20 mL顶空进样瓶中,孵育提取,进样。将孵育提取的挥发性风味物质释放到全二维气相色谱-飞行时间质谱(GC×GC-TOF MS)色谱系统中进行分析[8-10]
色谱条件:Agilent-8890A气相色谱仪(美国Agilent公司),以高纯氦气作为载气,恒定流速为1.0 mL/min。一维色谱柱DB-Heavy Wax (30 m×250 μm×0.5 μm)的初始温度为40 ℃,保持5 min,以5 ℃/min的速度升至100 ℃,随后以2 ℃/min的速度升至120 ℃,保持3 min,最后以6 ℃/min的速度升至250 ℃,保持5 min。二维色谱柱Rxi-5Sil MS(2 m×150 μm×0.15 μm)的升温程序高于一维色谱柱5 ℃,调制器温度始终高于二维色谱柱柱温15 ℃,调制周期为4.0 s。进样口温度为250 ℃。
质谱条件:使用LECO Pegasus BT 4D质谱质谱仪(美国LECO公司),质谱传输线温度为250 ℃,离子源温度为250 ℃,采集速率为200 spectra/s,电子轰击源为70 eV,检测器电压为2 036 V,质谱扫描范围为质荷比(m/z)35~550。

1.5 数据分析

采用SPSS 22.0软件对试验数据进行分析处理,结果用平均值±标准误(mean±SE)表示。采用t检验对2组数据的差异显著性进行分析,P<0.05表示差异显著;采用正交偏最小二乘-判别分析(OPLS-DA)对数据进行降维和可视化处理;采用Matlab软件对挥发性风味物质进行系统聚类分析。

2 结果与分析

2.1 吉富罗非鱼肌肉中挥发性风味物质分析

根据气相色谱仪测定出的挥发性风味物质的气相色谱保留时间和离子迁移时间,对挥发性风味物质进行定性分析。本试验共检测到920种挥发性风味物质,其中G0组检测出707种,G70组测出633种,2组共有的挥发性风味物质为420种(图1)。
图1 吉富罗非鱼肌肉中风味物质鉴定数目统计

Figure A:吉富罗非鱼肌肉中挥发性风味物质统计;图B:吉富罗非鱼肌肉中挥发性风味物质韦恩图。

Fig.1 Statistics on identification number of volatile flavor substances in muscle of GIFT

图A: statistics of volatile flavor substances in muscle of in GIFT; figure B: Venn diagram of volatile flavor substances in muscle of in GIFT.

2.2 吉富罗非鱼肌肉中挥发性风味物质的OPLS-DA

以2种吉富罗非鱼肌肉中挥发性风味物质含量原始数据矩阵进行OPLS-DA,结果描述了2组吉富罗非鱼肌肉中挥发性风味物质的主成分分布,分析组间的自然聚类趋势可发现,2组吉富罗非鱼肌肉中挥发性风味物质的主成分之间存在离散性(图2),主成分1(PC1,数据矩阵中最显著的特征)的贡献率为23.9%,正交成分2(OC2,正交与PC1的分类维度)的贡献率为49.6%,2个成分累积的贡献率为73.5%,这表明PC1和OC2的总贡献率包含了吉富罗非鱼肌肉样品的大部分信息,能够代表2组吉富罗非鱼肌肉样本中挥发性风味的主要特征。由图2可知,罗非鱼肌肉样本被分为2簇,G0组样本位于PC1的左侧,G70组样本位于PC1的右侧,OPLS-DA结果表明2组吉富罗非鱼肌肉中挥发性风味成分存在显著差异。
图2 吉富罗非鱼肌肉中挥发性风味物质的正交偏最小二乘-判别分析

Fig.2 OPLS-DA of volatile flavor substances in muscle of GIFT

表2 吉富罗非鱼肌肉中挥发性风味物质的差异分析

Table 2 Difference analysis of volatile flavor substances in muscle of GIFT %

项目
Items
组别Groups
G0 G70
(Z)-6-十一烯-2-酮C11H20O 0.226±0.061b 0.035±0.006a
1-辛烯-3-醇C8H16O 0.712±0.092b 0.330±0.019a
1-甲基吡咯C5H7N 0.293±0.010b 0.017±0.002a
5-甲基-2-己酮C7H14O 0.205±0.035b 0.031±0.004a
2-羟基-3-戊酮C5H10O2 0.131±0.006b 0.051±0.001a
6-甲基-2,4-庚二酮C8H14O2 0.019±0.002a 0.044±0.003b
6-甲基-5-庚烯-2-酮C8H14O 0.061±0.003b 0.001±0.000a
苯乙酸乙酯C10H12O2 0.539±0.023b 0.184±0.011a
异戊酸乙酯C7H14O2 0.747±0.098a 1.811±0.077b
1-巯基-2-丙酮C3H6OS 0.009±0.009a 0.209±0.021b

同行数据肩标不同小写字母表示差异显著(P<0.05)。

In the same row, values with different small letter superscripts mean significant difference (P<0.05).

2.3 吉富罗非鱼肌肉中挥发性风味物质差异分析

表2图3-a可知,G0组与G70组呈现出显著差异的挥发性风味物质有10种,分别为6-甲基-5-庚烯-2-酮、5-甲基-2-己酮、苯乙酸乙酯、1-辛烯-3-醇、1-甲基吡咯、(Z)-6-十一烯-2-酮、2-羟基-3-戊酮、6-甲基-2,4-庚二酮、异戊酸乙酯和1-巯基-2-丙酮。与G0组相比,G70组的6-甲基-5-庚烯-2-酮、5-甲基-2-己酮、苯乙酸乙酯、1-辛烯-3-醇、1-甲基吡咯、(Z)-6-十一烯-2-酮、2-羟基-3-戊酮的含量显著降低(P<0.05),6-甲基-2,4-庚二酮、异戊酸乙酯、1-巯基-2-丙酮含量显著提高(P<0.05)。由表3图3-b可知,与G0组相比,G70组肌肉中醇类、醛类、羧酸类、酯类的相对含量明显提高,杂环化合物、烃类和酮类的相对含量明显降低。G0组中,醇类的相对含量最高(32.97%),其次是酮类(10.82%);G70组中也是醇类的相对含量最高(40.52%),其次是酯类(13.98%),说明蚕豆对吉富罗非鱼肌肉中挥发性风味物质组成有影响。
图3 吉富罗非鱼肌肉中挥发性风味物质差异分析

图A:吉富罗非鱼肌肉中差异挥发性风味物质含量热图;图B:吉富罗非鱼肌肉中风味物质种类的相对含量。G0-1、G0-2和G0-3为G0组的3个样本,G70-1、G70-2和G70-3为G70组的3个样本。

Fig.3 Difference analysis of volatile flavor substances in muscle of GIFT

Figure A: heat map of differential volatile flavor substance contents in muscle of GIFT; figure B: relative contents of f volatile flavor substance types in muscle of GIFT. G0-1, G0-2 and G0-3 were three samples from G0 group, and G70-1, G70-2 and G70-3 were three samples from G70 group.

表3 吉富罗非鱼肌肉中不同种类挥发性风味物质相对含量比较

Table 3 Comparison of relative contents of different categories of volatile flavor substances in muscle of GIFT %

组别
Groups
醇类
Alcohols
醛类
Aldehydes
羧酸类
Carboxylic
acids
酯类
Esters
杂环化合物
Heterocyclic
compounds
烃类
Hydrocarbons
酮类
Ketones
其他
Others
G0 32.97 1.11 1.32 6.96 4.29 5.42 10.82 37.09
G70 40.52 1.53 2.06 13.98 2.40 5.06 7.41 27.04

2.4 主要挥发性风味物质阈值分析及感官风味特征分析

对吉富罗非鱼肌肉中主要挥发性风味物质的阈值进行相对气味活度值(ROVA)分析,ROVA≥1.0说明该物质是关键香味物质。ROVA越高,说明该挥发性风味物质对吉富罗非鱼肌肉风味的影响越大[11]。由表3可知,G0组和G70组吉富罗非鱼肌肉的关键香味物质主要来自醛类、酮类和烃类,这3大类物质对吉富罗非鱼肌肉的风味贡献较大。由表4可知,G70组吉富罗非鱼肌肉中2,3-丁二酮、2-甲基丁醛、2-戊基呋喃的ROVA较高(分别为76.56、100.00和1.37),而G0组吉富罗非鱼肌肉中2,3-丁二酮、2-甲基丁醛、2-十一烷、2-戊基呋喃的ROVA较高(分别为94.21、70.08、4.16和1.48)。
表4 吉富罗非鱼肌肉中挥发性风味物质阈值的相对气味活度值分析

Table 4 ROAV analysis of volatile flavor substances in muscle of GIFT

挥发性风味物质
Volatile flavor substances
气味阈值
Threshold odor/(μg/L)
气味特征
Odor characteristic
相对气味活度值ROVA
G0组G0 group G70组G70 group
2,3-丁二酮C4H6O2 0.002 黄油味 94.21 76.56
2-甲基丁醛C5H10O 0.001 可可味、杏仁味 70.08 100.00
2-十一烷C11H24 0.004 橙子味、鲜味 4.16 0.87
2-戊基呋喃C9H14O 0.006 绿豆味、蔬菜味 1.48 1.37
食品的风味是由两部分组成,分别是可识别的味觉和嗅觉特性,以及不能单独识别特性的复合体。本试验基于数据库Flavordb[12]中获取的ROVA,对2组吉富罗非鱼肌肉的感官风味进行分析和比较,并针对风味物质特有的感官风味特征,采用igraph构建它们之间的网络图关系。由图4可知,与G0组相比,G70组吉富罗非鱼肌肉的水果味和鲜味略微降低,其他风味如甜味、苦味、花香味、脂肪味、木质味、坚果味、蜡味、青草味等与G0组相似。由图5可知,吉富罗非鱼肌肉共检出5种感官风味,涉及10种口味。
图4 吉富罗非鱼肌肉感官风味特征分析

Fig.4 Analysis of muscle sensory flavor characteristics of GIFT

图5 吉富罗非鱼肌肉感官风味特征与挥发性风味物质关联网络图

Fig.5 Correlation network diagram between muscle sensory flavor characteristics and volatile flavor substances of GIFT

3 讨论

3.1 吉富罗非鱼肌肉中挥发性风味物质差异分析

醇类通常通过脂质的氧化分解产生,醇类化合物中,不饱和醇通常具有比饱和醇低得多的风味阈值,因此其对食品的风味属性具有更显著的影响[13]。与G0组相比,G70组吉富罗非鱼肌肉中1-辛烯-3-醇的含量明显降低。1-辛烯-3-醇具有蘑菇味,并且被认为是肉中泥土气味的来源[14]。饲喂添加高水平蚕豆饲料的吉富罗非鱼肌肉中醇类相对含量提高,而1-辛烯-3-醇含量降低,说明饲料中添加高水平蚕豆可有效降低吉富罗非鱼的土腥味,这可能是普通罗非鱼和脆肉罗非鱼肌肉风味差异的原因。这与本课题组前期研究结果一致,前期研究表明,饲料中添加高水平蚕豆可显著降低吉富罗非鱼肌肉中土臭素(GSM)含量,从而降低罗非鱼肌肉的土腥味[15]
酮类是由脂肪酸降解产生的,作为β-酮酸氧化的产物,通常与奶油和水果风味相关[16]。有研究表明,酮类对猪肉的香味有很大贡献,是猪肉香味的主要来源[17]。酮类也是水产品风味物质的重要组成部分,且对腥味物质具有增强作用[18]。刘晓丽等[19]研究表明,近江牡蛎(Ostrea rivularis)的挥发性风味物质中酮类占14.29%,其中3-辛酮、1-辛烯-3-酮、2,3-戊二酮的气味贡献较大,使牡蛎具有水果香、花香。与G0组相比,G70组吉富罗非鱼肌肉中酮类相对含量整体降低,而6-甲基-2,4-庚二酮、1-巯基-2-丙酮的含量显著提高,表明饲料中添加高水平蚕豆影响了吉富罗非鱼肌肉风味物质,酮类相对含量整体降低可能会抑制吉富罗非鱼中腥味物质的作用,从而赋予了脆肉罗非鱼独特的风味特征。
烃类由长链脂肪酸的自发氧化产生[16],此外,烃类在某些条件下可以形成醛类和酮类,从而影响整体风味[20]。有研究表明,鱼中存在大量的烃类,并且由于高气味阈值而限制了其对总体香味的贡献[21]。虽然烃类的特征气味不明显,但其是芳香体系的重要组成部分,在香气的保持和协调方面起着关键作用[22]。在本试验条件下,与G0组相比,G70组烃类相对含量略微降低,可能是脆肉罗非鱼肌肉风味降低的潜在因素。
肉质口感特性一般与肌肉中纤维直径、纤维密度、胶原蛋白含量等组织学结构特性有关,而肌肉中的脂肪也可影响鱼类肉质的口感风味,脂肪可增加肉质的柔嫩感和风味的浓郁感[23]。肉类风味的形成与其脂肪含量密切相关,瘦肉赋予肉汤浓郁的香气,而脂肪使肉类具有独特的风味。酯类通常通过脂质代谢产生的羧酸和醇的酯化而产生,并且对鱼肉的风味特征有实质性贡献[24]。有研究报道,脂肪酸组成是反刍动物肉风味的决定因素[25]。猪肉中的各种脂肪酸氧化衍生的挥发性化合物影响肉质风味[26]。与G0组(6.96%)相比,G70组酯类的相对含量明显提高(13.98%),表明蚕豆对吉富罗非鱼肌肉风味变化有较大影响。因此,脆肉罗非鱼的特殊风味与脂质及脂肪酸的代谢有关。本课题组前期研究结果表明,饲料中添加高水平蚕豆影响吉富罗非鱼肌肉脂肪酸组成[15],这可能是导致脆肉罗非鱼肌肉风味差异的重要因素。
醛类主要通过脂质氧化和降解反应产生,具有较低的气味阈值,并对风味有很大贡献[27]。先前的研究表明,醛类因其感知阈值低且含量相对较大,被认为是肉类的关键挥发性风味物质,并且会赋予肉类醛风味,如青草味和脂肪味等[28-31]。与G0组(1.11%)相比,G70组肌肉中的醛类相对含量略微提高(1.53%)。Fu等[32]的研究结果表明,醛类的表达上调可能导致了脆肉鲩的特殊风味,与本研究结果相似。

3.2 主要挥发性风味物质阈值分析及感官风味特征分析

挥发性风味物质对总体香气的贡献由其浓度和气味阈值决定[33]。一般认为,ROAV<1,说明该风味物质对总体气味无显著作用;ROAV≥1,说明该风味物质可能对总体气味有直接影响;且在一定范围内,ROAV越大,说明该风味物质对总体气味贡献越大[34]。本试验条件下,吉富罗非鱼肌肉中2,3-丁二酮、2-甲基丁醛、2-十一烷和2-戊基呋喃的ROVA较高,其气味阈值分别为0.002、0.001、0.004和0.006 μg/L,其中G0组的2,3-丁二酮、2-甲基丁醛的ROVA分别为94.21和70.08,G70组的2,3-丁二酮、2-甲基丁醛的ROVA分别为76.56和100.00。这意味着在吉富罗非鱼肌肉中,与其他挥发性风味物质相比,2,3-丁二酮和2-甲基丁醛对总体风味的贡献更大,这2种化合物呈现出的是黄油和可可的香味。在本试验条件下,与G0组相比,G70组的2,3-丁二酮的ROVA降低,而2-甲基丁醛的ROVA升高。2,3-丁二酮被认为与黄油味和脂肪味有关[35],说明饲料中添加高水平蚕豆可使吉富罗非鱼肌肉脂肪风味发生变化,与Fu等[32]的研究结果相似。肉类产生水果风味是脂肪氧化、糖类反应及风味化合物相互作用的结果。肉类中的脂肪分子受热分解产生醛类、酮类、酯类等挥发性化合物,具有水果香气(如柑橘类水果的醛类、坚果的酯类);肉类中的糖类,如葡萄糖、果糖等,发生美拉德反应会产生焦糖化产物,具有焦糖、烤面包等香气,与水果的香甜味相似[36-37]。本试验结果显示,与G0组相比,G70组吉富罗非鱼肌肉的水果风味和鲜味略微降低,可能与肌肉的脂类、糖类代谢密切相关,水果风味降低可能与苯乙酸乙酯和3-甲基丁酸乙基有关,苯乙酸乙酯和3-甲基丁酸乙酯是酯类风味物质,具有甜蜜香气和果香气味。

4 结论

在本试验中,对挥发性风味物质的OPLS-DA显示,饲料中添加高水平蚕豆对吉富罗非鱼肌肉中挥发性风味物质组成产生了显著影响;饲料中添加高水平(70%)蚕豆提高了吉富罗非鱼肌肉中醇类和酯类的相对含量,提高了挥发性风味物质2-甲基丁醛的ROVA以及降低了2,3-丁二酮的ROVA,可使肌肉脂肪风味发生变化;此外,饲料中添加高水平蚕豆略微降低了吉富罗非鱼整体肌肉的水果风味,并可抑制腥味物质的作用,降低肌肉中的土腥味。
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