CN113017053A - 一种富含油酸和挥发性风味物质的羊骨汤及其制备方法 - Google Patents
一种富含油酸和挥发性风味物质的羊骨汤及其制备方法 Download PDFInfo
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- CN113017053A CN113017053A CN202110422782.8A CN202110422782A CN113017053A CN 113017053 A CN113017053 A CN 113017053A CN 202110422782 A CN202110422782 A CN 202110422782A CN 113017053 A CN113017053 A CN 113017053A
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- Prior art keywords
- bone soup
- enzymolysis
- sheep bone
- sheep
- lipase
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
本发明属于食品加工领域，为了提高羊骨汤的不饱和脂肪酸含量和风味物质含量，以及感官品质，为羊骨汤的工业化生产提供理论依据，公开了一种富含油酸和挥发性风味物质的羊骨汤及其制备方法。本发明是通过加入脂肪酶进行酶解，测定酶解物的pH值、白度，可溶性固形物含量，将酶解物甲酯化处理后利用固相微萃取‑气相色谱‑质谱联用(SPME/GC‑MS)分析酶解对羊骨汤特征香气成分的影响，利用气相色谱‑质谱联用(GC‑MS)测定脂肪酸的含量，通过九点标度法对羊骨汤感官品质进行评价，明确脂肪酶酶解对羊骨汤理化性质、脂肪酸和风味物质含量及感官品质的影响，以期为羊骨汤的工业化生产提供理论依据。
Description
技术领域
本发明属于食品加工领域，具体涉及一种富含油酸和挥发性风味物质的羊骨汤及其制备方法。
背景技术
羊骨营养物质丰富，其中蛋白质、脂肪的含量与肉类十分相似，蛋白质含量达11％～15％，脂肪含量为10％～12％，而且还含有钙、钠、镁、钾、铁等矿物元素和丰富的维生素。
为了充分利用羊骨资源，羊骨常作为原料来熬制羊骨汤。羊骨汤中含有一定量的脂肪，脂肪对羊骨汤的主要作用是：脂肪加热降解产生各种挥发性化合物，如烷烃、醛、醇、酯和羧酸等，这些物质有的进一步参与美拉德反应形成更为复杂的香气物质。
但目前的烹食方法不能有效利用羊骨营养，导致大量对身体有益的物质流失，且传统烹食方式感官品质也较差。
如果利用酶解技术将骨汤中的脂肪进行水解，不仅可得到更多的有益脂肪酸，有利于风味物质的进一步生成，提高羊骨汤的风味特征，而且可以降低骨汤中的脂肪含量，得到改良的骨汤产品。
发明内容
本发明的目的是为了提高羊骨汤的不饱和脂肪酸含量和风味物质含量，以及感官品质，为羊骨汤的工业化生产提供理论依据，进而提供了一种富含油酸和挥发性风味物质的羊骨汤及其制备方法。
本发明采用如下技术方案：
一种富含油酸和挥发性风味物质的羊骨汤及其制备方法，羊骨汤原材料包括羊碎骨和脂肪酶，制备过程如下：
1)熬制羊骨汤将羊骨：水按照1：3(w/w)比例放入蒸煮锅中熬制，过滤后制得羊骨汤；
2)羊骨汤酶解
骨汤中添加脂肪酶，在酶解罐中进行酶解，酶解温度为25-50℃，脂肪酶添加量为0.5％-1.0％，酶解时间不少于3h，反应完成后灭酶；
3)羊骨汤调味、分装和灭菌
加入盐、胡椒粉、鸡精、味精和I+G，搅拌均匀，进行15～20MPa均质；将均质好的羊骨汤进行分罐，并用压罐机压盖，最后灭菌。
述脂肪酶的酶解温度为35℃，pH 7.5，脂肪酶添加量为0.8％，酶解反应完成后于90℃下灭酶10min。
所述熬制羊骨汤过程中，先中火熬制3小时，将熬制好的羊骨汤出锅，在锅中再次加入3倍羊骨体积的水重复熬制，连续熬制3次，合并熬制好的骨汤，利用双层纱布过滤制得羊骨汤。
羊骨粒径为0.9～1.1cm。
所述脂肪酶的活力为20U/mg。
酶解后羊骨汤的PH值、白度值和可溶性固形物含量降低，酶解合格。
酶解后羊骨汤的脂肪酸含量升高，酶解合格。特别是油酸含量由33.25％升高至58.52％。
酶解后羊骨汤的醛类物质增加，酶解合格。其中醛类物质包括15种挥发性醛类，壬醛在所测醛类物质中含量最高。
本发明是通过加入脂肪酶对羊骨汤进行酶解，测定了酶解羊骨汤的pH值、白度，可溶性固形物含量，并利用固相微萃取-气相色谱-质谱联用(SPME/GC-MS)分析酶解对羊骨汤特征风味物质种类和含量的影响，利用气相色谱-质谱联用(GC-MS)测定了脂肪酸的含量，通过九点标度法分析了酶解对羊骨汤感官品质的影响。结果表明：酶解导致羊骨汤的pH值、白度、可溶性固形物含量下降，而脂肪酸含量显著升高，特别是油酸含量由33.25％升高至58.52％；风味物质分析可知，与未酶解产品比较，酶解使产品的挥发性物质种类增多，含量增加。特别是醛类物质含量增加4.02倍，其中壬醛、庚醛、辛醛、反式-2-辛烯醛、反式-2-壬烯醛和2-十一烯醛含量显著增加。感官评定可知，酶解羊骨汤在气味综合评价、味觉综合评价和口感综合评价的评分都高于未酶解羊骨汤，所以总体接受度显著高于未酶解羊骨汤。因此，通过脂肪酶酶解，羊骨汤不仅不饱和脂肪酸含量和风味物质含量显著提高，而且感官品质提高，是一种有效提高羊骨汤风味和营养的方法。
附图说明
图1为未酶解羊骨汤风味物质的得分贡献图；
图2为酶解羊骨汤风味物质的得分贡献图；
图3为两种羊骨汤的感官评定雷达图。
具体实施方式
本发明是通过常压蒸煮制得羊骨汤，加入脂肪酶进行酶解，测定酶解物的pH值、白度，可溶性固形物含量，将酶解物甲酯化处理后利用固相微萃取-气相色谱-质谱联用(SPME/GC-MS)分析酶解对羊骨汤特征香气成分的影响，利用气相色谱-质谱联用(GC-MS)测定脂肪酸的含量，通过九点标度法对羊骨汤感官品质进行评价，明确脂肪酶酶解对羊骨汤理化性质、脂肪酸和风味物质含量及感官品质的影响，以期为羊骨汤的工业化生产提供理论依据。
本发明所采用材料和仪器如下：
羊碎骨(颗粒直径为0.9～1.1cm)：由山西怀仁金沙滩肉业有限公司提供；
脂肪酶(活力20U/mg)：江苏一鸣生物科技有限公司；酚酞、乙醇、甲醛、NaOH等试剂均为分析纯：天津化学试剂一厂。
酸度计(PB-10)：瑞典Sartorius公司；电子恒温水浴锅(ZKW-4)：常熟市天量仪器有限责任公司；电子天平(LT 100型)：北京中兴伟业公司；离心机(ST40R)：德国TheromoScientific公司；气相色谱仪(7890A)，美国Agilent公司
实施例1
羊骨汤的熬制：
将冷冻羊碎骨清水解冻，用冷水浸泡换水3次，水清为止；将羊骨放入蒸煮锅中以羊骨：水＝1：3(w/w)比例熬制，中火熬制3小时；将熬制好的羊骨汤出锅，在锅中再次加入3倍羊骨体积的水重复熬制，连续熬制3次，合并熬制好的骨汤，双层纱布过滤。
羊骨汤酶解：
将骨汤中添加脂肪酶在酶解罐中进行酶解，脂肪酶温度25℃，pH 7.5，脂肪酶添加量0.5％，酶解时间3h，反应完成后于90℃下灭酶10min。
羊骨汤调味、分装和灭菌：
加入盐0.3％，胡椒粉0.02％，鸡精0.03％，味精0.03％，I+G 0.005％(均以骨汤质量为基准)，搅拌均匀，进行15～20MPa均质；将均质好的羊骨汤分装到马口铁罐，每罐净含量280ml，用压罐机压盖；121℃灭菌30分钟。
测得酶解后羊骨汤油酸的相对含量为51.63％。
实施例2
羊骨汤酶解过程中的脂肪酶温度35℃，脂肪酶添加量0.8％，其余过程与实施例1相同。测得酶解后羊骨汤油酸的相对含量为58.52％。
实施例3
羊骨汤酶解过程中的脂肪酶温度50℃，脂肪酶添加量1.0％，其余过程与实施例1相同。测得酶解后羊骨汤油酸的相对含量为45.31％。
由上述3个实施例可知，由于酶活性受温度影响较大，温度过高会导致酶失活，影响酶解效果，并最终影响脂肪酸含量,尤其是油酸的含量。试验分析得出脂肪酶的最适温度为35℃，脂肪酶添加量0.8％。
为了进一步验证酶解对羊骨汤品质的影响，做了关于酶解羊骨汤和未酶解羊骨汤的对比试验例，一种按实施2步骤进行酶解制得羊骨汤，另一种除了不进行酶解步骤外，其余配料以及过程完全相同。
两种羊骨汤的指标测得方法如下：
1、pH值的测定
参照GB-5009.237-2016的方法进行测定
2、色差值的测定
取骨汤样品均匀铺布在色差平皿底部，将色差仪进行白板校正后检测其亮度(L*)、红度(a*)和黄度(b*)值，计算白度值按式(1)计算。
3、可溶性固形物含量的测定
试样在105±2℃烘箱内，在大气压下烘干，直至恒重。测定烘干后的质量m1与试样质量m，按以下公式(2)计算固形物百分比。
固形物含量(％)＝m1/m×100％ (2)
4、脂肪酸含量的测定
样品脂肪提取：称取样品5g放入250ml烧瓶中，加入100mg焦性没食子酸和十七烷酸甲酯内标溶液1ml，加入几粒沸石，再加入2ml 95％乙醇，混匀。然后加入8.3mol/L的盐酸溶液10ml，混匀。将烧瓶放入80℃水浴中水解40min。每隔10min振荡一次烧瓶，使黏附在烧瓶壁上的颗粒物混入溶液中。水解完成后，取出烧瓶冷却至室温。最后加入10ml 95％乙醇，混匀。
脂肪水解完成后，将烧瓶中的水解液转移到分液漏斗中，用50ml乙醚冲洗烧瓶，用乙醚冲洗烧瓶盖子，冲洗液并入分液漏斗中，加盖，振摇5min，静置10min，将醚层提取液收集到250ml烧瓶中，按照以上步骤重复提取水解液3次，最后用乙醚冲洗分液漏斗，并收集到250ml烧瓶中后，挥发提取溶剂，残留物为脂肪提取物。
脂肪的皂化和脂肪酸的甲酯化：在盛有脂肪提取物的250ml烧瓶中加入2％氢氧化钠甲醇溶液8ml，连接回流冷凝器在80℃水浴上回流30min；从回流冷凝器上端加入15％三氟化硼甲醇溶液7ml，在80℃水浴中继续回流2min；用去离子水冲洗回流冷凝器。停止加热，从水浴上去下烧瓶，迅速冷却至室温，准确加入30ml正庚烷，振摇2min，再加入饱和氯化钠水溶液20ml，静置分层，吸取上层正庚烷提取溶液5ml，转移到30ml比色管中，加入大约3～5g无水硫酸钠，振摇1min，静置5min，吸取上层溶液到进样瓶中待测定。
气相色谱分析：使用Agilent 7890A气相色谱仪。色谱柱为DB-WAX 30m×0.25mm石英毛细管柱；固定相FFAP，检测器温度280℃，进样口温度270℃，柱温130℃持续1min，以6.5℃/min的速度升至170℃；再以2.75℃/min升至215℃。保持32min。载气：N2；分流比50∶1。用脂肪酸标准品的保留时间定性，用面积归一化法及内标定量法分别进行分析。
5、挥发性风味物质的测定
称取6.5g骨汤，加入1g食盐，混合均匀，于80℃恒温水浴锅中加热15min。将活化好的萃取头插入萃取瓶，80℃恒温富集吸附风味物质30min后插入GC进样口，250℃解吸5min，进行GC-MS分析。根据保留时间和CAS号鉴定风味化合物。
6、感官评价
选取10名骨汤感官评定经验丰富的小组成员对羊骨汤的品质进行评价。感官评价采用九点标度法对羊骨汤进行评价。小组成员对样品的外观综合评价、气味综合评价、味觉综合评价、口感综合评价和总体接受程度进行评定，并对个体特征进行评分，即白度、分层程度、稠度；香味、羊膻味；鲜味、余味；口感细腻程度、醇厚度。每个属性在非结构化范围评价中从0(感官最差)到9(感官最好)表达，最后以雷达图的形式呈现。
数据处理时，试验均设3次重复，结果用平均值±标准差表示，采用MicrosoftExcel 2007和Simca14.1软件对试验数据进行统计分析，Origin Pro 8.0绘图软件绘图，并应用Statistics 8.1中Turkey HSD程序进行显著性差异分析，显著水平为0.05。
结果分析如下：
1、两种羊骨汤理化指标测定结果：
表1为两种羊骨汤中pH值、白度及可溶性固形物含量的测定结果。由表1可知，纯羊骨汤的pH值为7.63，酶解之后显著降低为6.97(P<0.05)，这是因为在脂肪酶的作用下，脂质分解产生脂肪酸，酸的积累导致pH值的下降。
W值表示白度，酶解导致白度的下降，但是与未酶解羊骨汤比较，差异不显著(P＞0.05)。酶解之后，脂质含量降低，脂质特有的白色减弱，故白度值降低。经过酶解之后可溶性固形物含量下降为3.82％，这是因为酶解之后，脂质物质分解变成可挥发性的小分子化合物的缘故。
表1两种羊骨汤的理化指标
Table 1 The physicochemicalindexes of sheep bone soup
注：A-C字母不同表示同列数据差异显著(P<0.05)
2、两种羊骨汤中脂肪酸含量测定
表2为两种羊骨汤中脂肪酸含量的测定结果。由表2可知，两种羊骨汤中共检测出6种脂肪酸，包括5种饱和脂肪酸和1种不饱和脂肪酸。由表可知，酶解过程对羊骨汤中脂肪酸含量变化显著。未酶解的羊骨汤中十四酸、十五酸、十六酸和十八酸含量高于酶解骨汤，但是油酸含量显著低于酶解骨汤(P<0.05)。油酸为不饱和脂肪酸，因含有双键更易发生氧化作用，生成过氧化物，过氧化物进一步分解生成酮、醛、酸等挥发性羰基化合物，产生特有的香气。因此油酸对于进一步热反应产生的香气成分具有重要的作用。
表2两种羊骨汤中脂肪酸的相对含量(％)
Table 2 The relative content of fatty acid in sheep bone soup
注：-表示未检测到。
3、两种羊骨汤中风味物质含量的测定
表3为羊骨汤中挥发性风味物质的组成及含量。在未酶解羊骨汤和酶解羊骨汤中分别检测到20、37种挥发性物质，包括醇、醛、烷、烯、酮、酯、呋喃类物质，其中以醇、醛、酮类物质含量丰富。酶解羊骨汤与未酶解羊骨汤比较发现，酶解使骨汤的挥发性物质种类增多，含量增加。
醇类在骨汤独特风味形成过程中至关重要。醇类主要来源于糖代谢、脂质氧化、氨基酸脱羧等代谢过程，脂肪酶的作用增加了参与这些代谢反应的底物的数量和含量。如表3所示未酶解羊骨汤和酶解羊骨汤共含有14种醇类，其中有9种醇类物质只存在于酶解羊骨汤中：4-甲基环己醇(0.45％)、异戊醇(0.12％)、2-戊炔-1-醇(0.03％)、2-七萜醇(2.61％)、正庚醇(2.30％)、2-辛炔-1-醇(0.17％)、正辛醇(0.02％)、苯乙醇(0.02％)、1-壬醇(0.05％)。醇类物质含量和种类的差异使酶解结束后两种骨汤的风味显著不同。
醛类是骨汤中典型的风味物质，两种羊骨汤共含有16种醛类，其中酶解羊骨汤含有15种挥发性醛类。壬醛在所测醛类物质中含量最高，相对含量分别为7.94％(未酶解羊骨汤)和22.00％(酶解羊骨汤)。壬醛具有较低的气味阈值，通常被认为是加工肉制品的主要风味成分。据报道壬醛在柑橘油类中含量较高，具有较强的月桂香和甜橙味。己醛具有青草味和脂肪味，在酶解羊骨汤中相对含量为5.82％。酶解结束后，庚醛的相对含量由0.97％显著增加到7.70％，为骨汤提供了果香味、柑橘味和坚果味。辛醛的相对含量由2.59％显著增加到11.74％，为骨汤提供了甜橙香气。酶解羊骨汤中还发现3种烯醛(反式-2-辛烯醛2.14％、反式-2-壬烯醛7.62％和2-十一烯醛2.70％)的相对含量显著高于未酶解羊骨汤(0.53％，1.02％，0.88％)，为骨汤提供了脂肪香、醛香、肉香等独特香气。总体来看，经过酶解后醛类物质种类和含量都有所增加，与未酶解羊骨汤比较，酶解羊骨汤的醛类物质相对含量增加4.02倍。
脂肪酸烷氧自由基的断裂和氨基酸降解代谢过程是形成酮类物质的主要途径。2-庚酮和3-辛烯-2-酮在酶解羊骨汤中能检测到，但不存在于未酶解羊骨汤中。但由于烷酮类物质的风味阈值比较高，故对骨汤风味形成贡献较小。
除此之外，酶解还使羊骨汤还产生了3-甲基-1-丁醇硝酸酯、硬脂酸3-十八烷基氧丙基酯以及2-正戊基呋喃等物质，且2-正戊基呋喃具有肉香味和硫味。
表3羊骨汤中挥发性风味物质的组成及含量
Table 3 The content of volatile compounds in sheep bone soup
4、两种羊骨汤风味物质的主成分分析
为进一步分析挥发性风味物质对羊骨汤风味的影响，对挥发性风味物质的相对含量进行主成分分析。由表4可知，主成分1的特征值为1.74，其贡献率为43.6％，主成分2的特征值为1.30，其贡献率为32.6％，即前两个主成分可解释76.2％的原始风味组分信息。因其特征值均大于1，可以推断出主成分1、2会影响羊骨汤中的主要风味和香气，所提取的两个主成分能够较好地反映羊骨汤风味的构成。
表4两种羊骨汤中挥发性风味物质的主成分分析
Table 4 Principal component analysis offlavor substances from sheepbone soup
图1为未酶解羊骨汤风味物质的得分贡献图。由图1可知，在未酶解羊骨汤的主成分分析中，几种烷烃与主成分1、2的得分贡献率较高，但烷烃类化合物主要来源于脂肪酸烷氧基自由基的均裂，一般认为对食品风味的贡献较少。A2(5-甲基-1，5-己二烯-3-醇)、A3(异辛醇)、A4(9-十八烷醇)、A9(12-甲基-2，13-十八碳二烯-1-醇)、B6(癸醛)、B8(枯茗醛)、B7(2-十一烯醛)、C1(正十四烷)、C2(正十二烷)、C3(正十六烷)、E1(2-癸酮)、E3(2-己酮)等起到风味的正向调控作用。
图2为酶解羊骨汤风味物质的得分贡献图。由图2可知，经过酶解后，羊骨汤中A4(9-十八烷醇)、A6(4-甲基环己醇)、A8(2-戊炔-1-醇)、A11(22-辛炔-1-醇)、A13(苯乙醇)、B3(反式-2-辛烯醛)、B5(反式-2-壬烯醛)、B16(顺式-4，5-环氧-(E)-2-癸烯醛)、C4(甲烷)、C5(iso-丙基环丁烷)、D1(6-甲基-3-辛烯)、E6(3-辛烯-2-酮)、F1(3-甲基-1-丁醇硝酸酯)、F2(硬脂酸3-十八烷基氧丙基酯)有明显的正相关性。
5、两种羊骨汤感官品质评价
基于九点标度法对两种羊骨汤分别进行感官评定，图3为两种羊骨汤的感官评分。由图3可知，在分层程度、稠度、外观综合评价及醇厚度方面两种羊骨汤的评分无显著性差异(P>0.05)；酶解羊骨汤在香味、无羊膻味得分高于未酶解羊骨汤，故气味综合评价较高；酶解羊骨汤在鲜味、余味得分高于未酶解羊骨汤，故味觉综合评价较高。
由于酶解羊骨汤在气味综合评价、味觉综合评价和口感综合评价的评分都高于未酶解羊骨汤，所以总体接受度显著高于未酶解羊骨汤(P<0.05)，表明经过脂肪酶酶解的羊骨汤更受消费者的欢迎。
这可能是因为脂肪酶将脂肪分解成小分子物质，产生特殊的挥发性风味物质，有助于酶解羊骨汤特征风味的形成。一般羊骨汤有羊膻味的存在，但是酶解之后，羊膻味减弱，且具有特殊香味，使羊骨汤风味得到较大提高。
有关研究表明蛋白酶解会有一定的苦味，影响口感和风味，这是酶解过程促进了苦味肽的形成。而经过脂肪酶酶解，并未有苦味肽的产生，产生的是以醛、醇、酯和脂肪酸为代表的分子量较低的物质，这些物质进一步参与美拉德反应形成风味物质，不仅使羊膻味减弱，而且风味得到了较大改善，鲜香味浓郁，获得了较高的感官评分。
通过对酶解羊骨汤理化指标、脂肪酸含量和挥发性风味物质含量分析可知，脂肪酶酶解促进了油酸的形成和挥发性风味物质数量和含量的增加，油酸含量由33.25％升高至58.52％，风味物质中壬醛、庚醛、辛醛、反式-2-辛烯醛、反式-2-壬烯醛和2-十一烯醛含量显著增加，酶解羊骨汤的感官评分也显著高于未酶解产品。因此，利用脂肪酶酶解提高了羊骨汤的风味评分和感官品质。
Claims (8)
1.一种富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，羊骨汤原材料包括羊碎骨和脂肪酶，制备过程如下：
1)熬制羊骨汤
将羊骨：水按照1：3(w/w)比例放入蒸煮锅中熬制，过滤后制得羊骨汤；
2)羊骨汤酶解
骨汤中添加脂肪酶，在酶解罐中进行酶解，酶解温度为25-50℃，脂肪酶添加量为0.5％-1.0％，酶解时间不少于3h，反应完成后灭酶；
3)羊骨汤调味、分装和灭菌
加入盐、胡椒粉、鸡精、味精和I+G，搅拌均匀，进行15～20MPa均质；将均质好的羊骨汤进行分罐，并用压罐机压盖，最后灭菌。
2.根据权利要求1所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，所述脂肪酶的酶解温度为35℃，pH 7.5，脂肪酶添加量为0.8％，酶解反应完成后于90℃下灭酶10min。
3.根据权利要求2所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，所述熬制羊骨汤过程中，先中火熬制3小时，将熬制好的羊骨汤出锅，在锅中再次加入3倍羊骨体积的水重复熬制，连续熬制3次，合并熬制好的骨汤，利用双层纱布过滤制得羊骨汤。
4.根据权利要求3所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，羊骨粒径为0.9～1.1cm。
5.根据权利要求4所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，所述脂肪酶的活力为20U/mg。
6.根据权利要求5所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，酶解后羊骨汤的pH值、白度值和可溶性固形物含量降低，酶解合格。
7.根据权利要求6所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，酶解后羊骨汤的脂肪酸含量升高，酶解合格。
8.根据权利要求7所述的富含油酸和挥发性风味物质的羊骨汤及其制备方法，其特征在于，酶解后羊骨汤的醛类物质增加，酶解合格。
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