摘要
为探究臭氧处理对冷冻鱼糜保质期内的蛋白氧化和风味影响,以鲢为原料,采用臭氧漂洗制备新鲜和冷冻的鱼糜,分别测定盐溶性蛋白、水溶性蛋白、活性巯基、游离氨基酸含量,并采用电子鼻、电子舌和气相色谱-离子迁移谱联用仪(GC-IMS)测定挥发性风味成分组成。结果显示,臭氧漂洗新鲜鱼糜的水溶性蛋白含量下降,盐溶性蛋白含量上升;冷冻贮藏后,水溶性、盐溶性蛋白含量均显著下降(P<0.05)。臭氧漂洗新鲜鱼糜活性巯基含量显著下降(P<0.05),冷冻贮藏后,活性巯基含量也有显著下降,但与臭氧漂洗相比,下降幅度较小。臭氧漂洗鱼糜的游离氨基酸含量显著下降,冷冻贮藏后,游离氨基酸含量大幅度下降。电子鼻、电子舌检测结果显示,冷冻鱼糜与新鲜鱼糜的气味、滋味可以有效区分开,但臭氧漂洗鱼糜与清水漂洗鱼糜不能有效区分。GC-IMS共鉴定出59种挥发性风味物质,臭氧漂洗鱼糜检测出的不饱和醇类数量明显低于自来水漂洗鱼糜,并且在冷冻后醇类物质减少,醛类物质显著增加。结果表明,臭氧漂洗初步运用到鱼糜及鱼糜制品等水产加工业中,可以改善鱼糜品质及风味,但在鱼糜冷冻贮藏过程中影响较小。
鱼糜是我国传统水产制品,具有高蛋白、低脂肪、营养价值高、安全健康等优点,深受消费者欢迎,2021年我国鱼糜制品产量达到134.98万t,相较2020年增加6.47
漂洗是制作鱼糜的一道关键工序,漂洗可以富集肌原纤维蛋白,提高其浓度,增强鱼糜凝胶形成能力,去除鱼肉腥味,改善鱼糜品质,同时也会降低一些呈味核苷酸类物质和水溶性游离氨基酸的含量,进而影响鱼糜产品的风味特
本研究以鲢为原料,采用臭氧漂洗制备新鲜和冷冻的鱼糜,通过测定盐溶性蛋白、水溶性蛋白、活性巯基含量、游离氨基酸组成以及电子鼻、电子舌和GC-IMS检测挥发性风味成分等方法研究臭氧漂洗及冷冻贮藏对鱼糜蛋白氧化和风味影响,分析蛋白氧化与风味物质变化的内在联系,研究臭氧漂洗工艺对新鲜鱼糜及冷冻鱼糜风味的影响,旨在为臭氧在鱼糜及鱼糜制品等水产加工业中的应用提供理论基础,为优质鱼糜的生产和品质调控提供方法和依据。
新鲜鲢(2.0~3.0 kg/尾),购于华中农业大学校内农贸市场,活鱼运输到实验室。
GCQJ-1-3电解式高浓度臭氧气机,武汉威蒙环保科技有限公司;ASTREE II电子舌,Alpha M.O.S公司;L-8900全自动氨基酸分析仪,日本日立公司;FOX-4000型电子鼻,Alpha M.O.S公司;7890A气相色谱仪,美国安捷伦公司;FlavourSpe
新鲜鲢清洗,去头、去尾去内脏后用采肉机采肉,均分为2组,第1组用清水(自来水)漂洗2次;第2组用清水漂洗1次,8 mg/L臭氧水漂洗1次,漂洗完鱼肉用三足式离心机离心脱水,再使用水分分析仪测定并调节水分含量至76%,得到清水漂洗新鲜鱼糜(QSYM)和臭氧漂洗新鲜鱼糜(O3YM)。在上述2种鱼糜中加入抗冻剂(4%蔗糖、4%山梨糖醇、0.1%焦磷酸钠、0.1%三聚磷酸钠),使用食品调理机斩拌均匀,放置于-18 ℃冷冻贮藏45 d,得到清水漂洗冷冻鱼糜(QSLD)和臭氧漂洗冷冻鱼糜(O3LD)。
1)水溶性及盐溶性蛋白的测定。参考Xiong
2)羰基含量的测定。参考Mesquita
3)活性巯基含量的测定。参考Zhang
4)游离氨基酸含量测定。参考GB 5009.124-2016《食品中氨基酸的测定》方法。
取2 g样品置于20 mL顶空瓶中,60 ℃孵育20 min后进样,进样针温度65 ℃,进样500 μL,设置分析条件:MXT-5色谱柱(15 m×0.53 mm,1 μm),柱温60 ℃,使用N2作为载气和漂移气,漂移气流速为150 mL/min,载气流速在0~2 min为2 mL/min,2~20 min内为100 mL/min,每个样品重复测定3次。采用系统自带的VOCal、Reporter插件、GalleryPlot插件和DynamicPCA插件进行谱图和数据的定性定量以及二维俯视图、三维谱图和差异谱图、指纹图谱的分析。
由

图1 不同处理鲢鱼糜的水溶性蛋白与盐溶性蛋白含量
Fig.1 The contents of water-soluble protein and salt-soluble protein in surimi of silver carp under different treatments
QSYM表示清水漂洗新鲜鱼糜;O3YM表示臭氧漂洗新鲜鱼糜;QSLD表示清水漂洗冷冻鱼糜;O3LD代表臭氧漂洗冷冻鱼糜。不同大写字母表示水溶性蛋白含量差异显著,小写字母表示盐溶性蛋白含量差异显著(P<0.05)。QSYM represents tag water rinsing fresh surimi; O3YM represents ozone water rinsing fresh surimi; QSLD represents tag water rinsing frozen surimi; O3LD represents ozone water rinsing frozen surimi. Different capital letters indicate significant differences in water-soluble protein content, while small letters indicate significant differences in salt soluble protein content (P<0.05).
由

图2 不同处理鲢鱼糜的羰基(A)及活性巯基含量(B)
Fig. 2 The contents of carbonyl(A) and active sulfhydryl content(B) in surimi of silver carp under different treatments
不同字母表示差异显著(P<0.05)。Different letters indicate a significant difference (P<0.05).
滋味活度值(taste activity value,TAV)为样品中滋味物质浓度与其阈值的比值,当TAV≥1时,表明该滋味物质对样品滋味贡献较大。由
氨基酸 Amino acid | 阈值/(mg/100 g) Threshold | 滋味 Taste | QSYM | O3YM | QSLD | O3LD | ||||
---|---|---|---|---|---|---|---|---|---|---|
含量/(mg/100 g) Content | TAV | 含量/(mg/100 g) Content | TAV | 含量/(mg/100 g) Content | TAV | 含量/(mg/100 g) Content | TAV | |||
天冬氨酸Asp | 100 | 鲜/甜 Delicate/sweet(+) | 1.571±0.211a | 0.015 7 | 1.052±0.300a | 0.010 5 | 0.551±0.037b | 0.005 5 | 0.813±0.140b | 0.008 1 |
苏氨酸Thr | 260 | 甜 Sweet(+) | — | — | — | — | 1.413±0.168a | 0.005 4 | 1.733±0.247a | 0.006 7 |
丝氨酸Ser | 150 | 甜 Sweet(+) | 0.510±0.096c | 0.003 4 | 0.528±0.066c | 0.003 5 | 1.058±0.033b | 0.007 1 | 1.309±0.114a | 0.008 7 |
谷氨酸Glu | 30 | 鲜 Delicate(+) | 32.946±4.523a | 1.098 2 | 21.894±3.296b | 0.729 8 | 5.509±0.305c | 0.183 6 | 7.482±0.906c | 0.249 4 |
甘氨酸Gly | 130 | 甜 Sweet(+) | 7.316±0.236a | 0.056 3 | 4.825±0.470b | 0.037 1 | 0.748±0.004c | 0.005 8 | 0.368±0.046d | 0.002 8 |
丙氨酸Ala | 60 | 甜 Sweet(+) | 28.799±0.930a | 0.480 0 | 15.297±0.009b | 0.255 0 | 2.368±0.242c | 0.039 5 | 3.185±0.460c | 0.053 1 |
胱氨酸Cys | — | 苦/甜/硫 Bitter/sweet/sulfur(-) | 9.088±0.265a | — | 6.803±0.192b | — | 0.010±0.001c | — | — | — |
缬氨酸Val | 40 | 甜/苦 Sweet/bitter(-) | 20.354±2.405a | 0.508 9 | 15.366±0.880b | 0.384 2 | 2.679±0.114c | 0.067 0 | 3.326±0.365c | 0.083 2 |
蛋氨酸Met | 30 | 苦/甜/硫Bitter/sweet/sulfur(-) | 1.274±0.093b | 0.042 5 | 1.091±0.073b | 0.036 4 | 1.276±0.057b | 0.042 5 | 2.015±0.220a | 0.067 2 |
异亮氨酸Ile | 90 | 苦 Bitter(-) | 12.163±0.776a | 0.135 1 | 8.565±0.952b | 0.095 2 | 0.232±0.017c | 0.002 6 | 0.299±0.032c | 0.003 3 |
亮氨酸Leu | 190 | 苦 Bitter(-) | 22.963±2.487a | 0.120 9 | 14.098±0.577b | 0.074 2 | 0.866±0.088c | 0.004 6 | 0.853±0.052c | 0.004 5 |
酪氨酸Tyr | — | 苦 Bitter(-) | 3.782±0.150a | — | 3.532±0.077b | — | — | — | 0.096±0.005c | - |
苯丙氨酸Phe | 90 | 苦 Bitter(-) | 12.084±0.294a | 0.134 3 | 9.139±0.954b | 0.101 5 | 0.804±0.257c | 0.008 9 | 0.993±0.078c | 0.011 0 |
赖氨酸Lys | 50 | 甜/苦 Sweet/bitter(-) | 18.539±0.251a | 0.370 8 | 15.707±1.093b | 0.314 1 | 4.626±0.499d | 0.092 5 | 7.690±1.311c | 0.153 8 |
组氨酸His | 20 | 苦 Bitter(-) | 8.043±0.589a | 0.402 2 | 5.080±0.259b | 0.254 0 | 9.301±0.167a | 0.465 1 | 7.144±1.405ab | 0.357 2 |
精氨酸Arg | 50 | 甜/苦 Sweet/bitter(+) | — | — | — | — | 1.492±0.101b | 0.029 8 | 2.654±0.938a | 0.053 1 |
脯氨酸Pro | — | 甜/苦 Sweet/bitter(+) | — | — | — | — | — | — | — | — |
TUSAA | 71.142±5.102a | 43.596±3.523b | 13.170±0.359c | 17.595±2.923c | ||||||
TBAA | 108.291±1.059a | 79.380±0.336b | 19.794±0.362d | 22.416±0.657c | ||||||
TFAA | 179.433±6.161a | 122.977±3.187b | 32.964±0.721c | 40.010±3.580c |
注: TUSAA表示甜味氨基酸 ,TBAA表示苦味氨基酸, TFAA表示游离氨基酸。Note: TUSAA as sweet amino acid, TBAA as bitter amino acid, TFAA as free amino acid.
对电子舌采集到的各组数据进行主成分分析,并做主成分分析(PCA)图和滋味雷达图。由

图3 不同处理鲢鱼糜基于电子舌滋味特征的PCA分析(A)和雷达图(B)
Fig.3 PCA analysis (A) and radar chart (B) of electronic tongue taste characteristics of silver carp surimi under different treatments
由

图4 鲢鱼糜基于电子鼻气味特征的PCA分析(A)、雷达图(B)和腥味评价(C)
Fig.4 PCA analysis (A), radar chart (B) and odor evaluation (C) of silver carp surimi based on odor characteristics of electronic nose
由4种鱼糜的电子鼻响应值的雷达图(
从GC-IMS三维俯视图(

图5 鱼糜挥发性组分的GC-IMS谱图俯视图(A)、差异图(B)和指纹图谱(C)
Fig.5 GC-IMS spectrum top view (A), two-dimensional differential map (B) and fingerprint map (C) of the volatile components of surimi of silver carp
C图中横坐标数字代表未能定性物质。The abscissa number in Fig.C represents the non qualitative substance.
GC-IMS从样品中共鉴定出挥发性风味物质59种,其中醛类17种、醇类13种、酮类7种、酯类1种、其他化合物21种,其中有2-戊基呋喃等重要风味化合物。清水漂洗新鲜鱼糜壬醛、辛醛、6-甲基-5-庚-2-酮、1-戊醇、醋酸、异丙醇等物质的含量较高,臭氧漂洗后的新鲜鱼糜3-辛醇、庚醛、1-戊醇、戊醛、3-甲基丁醛、异丙醇、β-罗勒烯、庚醇等物质的含量较高,清水漂洗冷冻鱼糜3-辛酮、1-辛烯-3-醇、3-呋喃甲醇、环己酮、(E)-2-戊烯醛、2-甲基-1-丁醇、3-甲基-1-丁醇、3-戊酮、1-丁醇、乙酸乙酯、丁醛、丙醇、2-丁酮、糠醛、异丁醇等物质的含量较高,其中醇类、烯醛类香气物质的含量较高,大多是鱼腥味的重要特征物质。臭氧漂洗后的冷冻鱼糜中(E)-2-己烯醛、乙偶姻、2-甲基丁醛、3-甲基丁醛、1-丁醇、丙醇、2-丁酮等物质的含量较高。
由
鱼肉主要成分一般由水溶性的肌浆蛋白、盐溶性的肌原纤维蛋白和不溶性的基质蛋白组成,已有研究表明臭氧漂洗会一定程度改变这些鱼肉蛋白结构,进而导致蛋白溶解度发生一定程度的改变,适度的臭氧氧化导致鱼肉蛋白结构发生轻微改变,增强了蛋白的溶解性,使其在漂洗过程中溶出增
鱼肉中丰富的游离氨基酸是其滋味特性的主要来源之一。现有的研究表明,鲢鱼肉特有的风味游离氨基酸主要是组氨酸、赖氨酸、丙氨酸、苏氨酸等,而鲢鱼糜中苦味的亮氨酸、异亮氨酸、苯丙氨酸、缬氨酸、蛋氨酸,以及具有鲜味的谷氨酸的含量相对较高,且谷氨酸对生鱼糜滋味贡献较
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