CN112881622B - 涩感物质影响程度的判断方法 - Google Patents
涩感物质影响程度的判断方法 Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N15/02—Investigating particle size or size distribution
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Abstract
本发明提出一种涩感物质影响程度的判断方法,属于啤酒酿造技术领域。该方法利用唾液沉淀指数结合粒径大小、粘度、摩擦系数等对涩感影响物质程度提供判断方法,可在筛得影响啤酒涩感物质的基础上,进一步对其进行量化,相比于传统感官品评方式更加客观、准确;此外,基于上述可量化的结果可根据不同啤酒产品的特点制定个性化调控技术,保证不同品类和风格的啤酒达到不涩的要求。
Description
技术领域
本发明属于啤酒酿造技术领域,尤其涉及一种涩感物质影响程度的判断方法。
背景技术
涩感是一个多因素综合影响的口感感官指标,影响因素主要包括植物单宁、多酚、有机酸、多价阳离子、脱水剂以及矿物质等。目前的报道中多酚是涩感的主要影响物质,但并不是所有的多酚都会带来影响,其中只有富含Galloyl基团的多酚是最容易与唾液蛋白发生反应,从而带来涩感。
目前的研究发现,红酒中影响涩感的因素为乳酸和苹果酸,而关于啤酒的涩感研究报道中仅报道了麦芽中的生物碱、蛋白和香气对涩感感官的影响。但问题在于啤酒中包含糖、有机酸、醇酯、离子、蛋白、多酚等多种物质,到底哪些物质会带来涩感,哪些物质会掩盖涩感尚不清楚。为了了解其内在影响,我们采用传统的感官品评,但感官品评并无法量化;也试图采用其他酒类、植物类的涩感研究方法,但啤酒组分具有特殊性,影响涩感的物质除了涩感物质本身外,还受到啤酒基质的影响,例如同样的多酚在红酒是涩的,在啤酒中可能不涩。因此,尚未有一种合理、可量化的方法可以判断啤酒中究竟哪些物质会带来涩感、哪些物质会掩盖涩感以及其影响程度究竟是怎样的。
发明内容
本发明提供了一种涩感物质影响程度的判断方法,该方法利用唾液沉淀指数结合粒径大小、粘度、摩擦系数等因素为涩感物质影响程度提供判断方法,可在筛得影响啤酒涩感物质的基础上,进一步对其进行量化,相比于传统感官品评方式更加客观、准确。
为了达到上述目的,本发明提供了一种涩感物质影响程度的判断方法,包括以下步骤:
利用偏最小二乘回归法建立啤酒唾液蛋白沉淀指数和啤酒8大类物质组分之间的回归方程,筛选回归方程中标准化系数绝对值大于0.3的组分,以该组分作为影响啤酒涩感的可能性啤酒组分;
将上述筛选得到的可能性啤酒组分按照啤酒浓度的10倍分别配制单物质溶液,通过分别与唾液进行沉淀反应,将可能性啤酒组分初步分为涩感因素和掩涩因素,其中,涩感因素是与唾液发生沉淀反应带来涩感的物质,掩涩因素是降低摩擦系数、提高润滑性能,可以掩盖涩感的物质;
根据不同组分与唾液反应的沉淀指数的增加幅度以及粒径大小判断属于涩感因素的各组分的最终影响程度,以及根据粘度与摩擦系数的变化程度判断属于掩涩因素的各组分的最终影响程度。
作为优选,所述8大类物质包括有机酸、阴离子和阳离子、多糖、蛋白、多酚及涩感多酚、醇酯、苦味酸和粘度。
作为优选,利用偏最小二乘回归法建立啤酒唾液蛋白沉淀指数和啤酒8大类物质组分之间的回归方程具体为:
Y=-370+0.078ⅹ乳酸-1.52ⅹ琥珀酸/苹果酸+0.709ⅹ柠檬酸+0.726ⅹ草酸+0.377ⅹ丙酮酸-0.02ⅹ富马酸-0.04ⅹ乙酸+0.032ⅹ总有机酸+0.004ⅹK++229ⅹMn2++0.106ⅹNa++34.7ⅹBa2+-0.686ⅹCa2+-104ⅹAl3++0.378ⅹMg2+-0.0711ⅹ总阳离子-0.0018ⅹCl--0.0008ⅹPO4 3--0.3ⅹSO4 2-+0.056ⅹ涩感多酚+0.0028ⅹ总醇-0.02ⅹ总酯+0.103ⅹ苦味酸-0.22ⅹ多糖-0.015ⅹ蛋白+330ⅹ粘度-0.004ⅹ异戊醇+0.06ⅹ乙酸乙酯。
作为优选,标准化系数绝对值大于0.3的组分包括选自乳酸、琥珀酸、苹果酸、柠檬酸、丙酮酸的有机酸,选自Ca2+、Mg2+、K+、Al3+的阳离子,选自SO4 2-、PO4 3-、Cl-的阴离子以及涩感多酚、多糖、蛋白和粘度。
作为优选,分别与唾液进行沉淀反应后,依据沉淀指数是否>2%将可能性啤酒组分分为与唾液发生沉淀反应的组分和与唾液无沉淀反应的组分。
作为优选,对与唾液发生沉淀反应的组分进行粒径分析,将粒径>500nm的组分初判为涩感因素;对与唾液无沉淀反应的组分进行粘度分析,将粘度增加的组分初判为掩涩因素。
作为优选,所述涩感因素包括乳酸、琥珀酸、苹果酸、柠檬酸、Ca2+、SO4 2-和涩感多酚;所述掩涩因素包括多糖和蛋白,其中所述多糖包括淀粉多糖和非淀粉多糖。
作为优选,将得到的涩感因素分别以啤酒浓度的50%反添加入啤酒中,根据沉淀指数增加幅度以及粒径大小判断涩感因素的最终影响程度具体为:
对于粒径500-1000nm的组分,其沉淀指数的增加幅度<10%,定义为III类;>10%,定义为II类;
对于粒径1000-2000nm的组分,其沉淀指数的增加幅度<5%,定义为III类;5%-10%,定义为II类;>10%,定义为I类;
对于粒径>2000nm的组分,其沉淀指数的增加幅度<5%,定义为II类;>5%,定义为I类;
其中,I类为影响程度大;II类为影响程度中等;III类为影响程度小。
作为优选,涩感因素的最终影响程度具体为:
作为优选,向啤酒中以万分之一体积比外添加不同的专一性酶制剂分别降解属于掩涩因素的各组分,其中,对应淀粉多糖加入的是葡糖淀粉酶,对应非淀粉多糖加入的是葡聚糖酶和木聚糖酶的复合物,对应蛋白加入的是蛋白酶;
掩涩因素的最终影响程度为非淀粉多糖、淀粉多糖和蛋白。
与现有技术相比,本发明的优点和积极效果在于:
本发明利用唾液沉淀指数结合粒径大小、粘度、摩擦系数等因素对涩感物质影响程度提供判断方法,可在筛得影响啤酒涩感物质的基础上,进一步对其进行量化,相比于传统感官品评方式更加客观、准确;此外,基于上述可量化的结果可根据不同啤酒产品的特点制定个性化调控技术,保证不同品类和风格的啤酒达到不涩的要求。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本发明实施例提供了一种涩感物质影响程度的判断方法,包括以下步骤:
S1:利用偏最小二乘回归法建立啤酒唾液蛋白沉淀指数和啤酒8大类物质组分之间的回归方程,筛选回归方程中标准化系数绝对值大于0.3的组分,以该组分作为啤酒涩感的可能性啤酒组分;
S2:将上述筛选得到的可能性啤酒组分按照啤酒浓度的10倍分别配制单物质溶液,通过分别与唾液进行沉淀反应,将可能性啤酒组分初步分为涩感因素和掩涩因素;
S3:根据不同组分与唾液反应的沉淀指数的增加幅度以及粒径大小判断属于涩感因素的各组分的最终影响程度,以及根据粘度以及摩擦系数的变化程度判断属于掩涩因素的各组分的最终影响程度。
上述步骤S1中,通过对25个同等度数(8P啤酒)的啤酒样品进行分析,在一优选实施例中,所述8大类物质包括有机酸、阴离子和阳离子、多糖、蛋白、多酚及涩感多酚、醇酯、苦味酸和粘度。具体如下:
1、唾液蛋白沉淀指数及啤酒多酚沉淀指数分析
1.1唾液收集:唾液分泌受人体昼夜生理规律的影响,呈现一定的波动变化,固定下午14:00-16:30之间收集。唾液采集前1h内不能进食,采集当天不能饮酒、喝含咖啡因饮料。采集前半小时喝室温饮用水200mL,然后收集1h唾液,混匀,冰箱4度放置过夜,然后12000rpm离心10min取上清。
1.2唾液与啤酒样品反应:唾液与啤酒按照1:1比例在37度反应10min,转速650rpm~750rpm,反应后12000rpm离心10min,取上清;同时设定唾液对照和样品对照,如表1所示。
表1样品及对照品制备
啤酒样品 | 唾液 | 水 | |
唾液对照1-1 | 1mL | 1mL | |
唾液对照1-2 | 1mL | 1mL | |
唾液对照1-3 | 1mL | 1mL | |
啤酒样品对照1-1 | 1mL | 1mL | |
啤酒样品对照1-2 | 1mL | 1mL | |
啤酒样品对照1-3 | 1mL | 1mL | |
唾液&啤酒样品反应样品1-1 | 1mL | 1mL | |
唾液&啤酒样品反应样品1-2 | 1mL | 1mL | |
唾液&啤酒样品反应样品1-3 | 1mL | 1mL |
1.3蛋白检测方法(考马斯亮蓝分析方法):
1)标准曲线的制备:用0.1mg/mL标准蛋白质BSA溶液配制不同浓度梯度的标准溶液,梯度稀释的标准溶液体积为0.5mL。加入5mL考马斯亮蓝染色液,用漩涡振荡器混匀液体,静置5min。在595nm下检测液体的吸光度。以蛋白质质量(mg)为纵坐标,以测得的吸光度为横坐标,绘制标准曲线,建立回归方程Y=0.0028x+0.2794,R2>0.99,计算样品OD值=0.0028ⅹ蛋白浓度+0.2794。
2)取样品0.15mL,补加0.15mol/L NaCl溶液0.35mL,使混合后的液体体积为0.5mL。加入5mL考马斯亮蓝染色液,用漩涡振荡器混匀液体,静置5min。在595nm下检测试液的吸光度。通过标准曲线换算样品蛋白浓度,高分子蛋白含量(mg/L)=(样品OD值-0.2794)ⅹ10/3/0.0028。
1.4总多酚测定方法:
1)取两个25mL容量瓶,分别标识空白样品B、检测样品A,分别加入10mL样品,再加入8mL羧甲基纤维素溶液,充分混匀;
2)将检测样品内加入0.5mL铁试剂,充分混匀;
3)将空白样品、检测样品内分别加入0.5mL氨水(1:2),用蒸馏水稀释至刻度,充分混匀;
4)将空白样品和检测样品于常温下静置10分钟后,用10mm玻璃比色皿,分别测定空白样品B和检测样品A的吸光度;
5)总多酚(mg/L)=(A600-B600)ⅹ820。
1.5沉淀指数计算方法:
唾液蛋白沉淀指数(%)=(唾液对照+啤酒样品对照-唾液&啤酒样品试验)*100/唾液对照;
啤酒多酚沉淀指数(mg/L)=唾液对照+啤酒样品对照-唾液&啤酒样品试验
2、啤酒组分分析方法:
采用体积排阻液相色谱法测定啤酒单糖、二糖、三糖及四糖以上组分的含量;
采用考马斯亮蓝法测定啤酒中高分子蛋白的含量;
采用离子色谱法测定SO4 2-、PO4 3-、Cl-、柠檬酸、富马酸、草酸、琥珀酸/苹果酸、丙酮酸、甲酸、乙酸、乳酸的含量;
采用电感耦合等离子体发射光谱仪测定K+、Na+、Ca2+、Mg2+的含量;
采用气相色谱法测定异戊醇、异丁醇、正丙醇、辛酸乙酯、己酸乙酯、乙酸异戊酯、乙酸乙酯的含量;
采用液相色谱法测定啤酒中的苦味酸含量;
采用安东帕粘度分析仪分析粘度,利用安东帕的MCR摩擦磨损分析仪。测定摩擦系数。
3、通过PLSR方程建立预测模型
利用偏最小二乘回归法建立啤酒多酚沉淀指数和啤酒8大类物质组分之间的回归方程具体为:
Y=-370+0.078ⅹ乳酸-1.52ⅹ琥珀酸/苹果酸+0.709ⅹ柠檬酸+0.726ⅹ草酸+0.377ⅹ丙酮酸-0.02ⅹ富马酸-0.04ⅹ乙酸+0.032ⅹ总有机酸+0.004ⅹK++229ⅹMn2++0.106ⅹNa++34.7ⅹBa2+-0.686ⅹCa2+-104ⅹAl3++0.378ⅹMg2+-0.0711ⅹ总阳离子-0.0018ⅹCl--0.0008ⅹPO4 3--0.3ⅹSO4 2-+0.056ⅹ涩感多酚+0.0028ⅹ总醇-0.02ⅹ总酯+0.103ⅹ苦味酸-0.22ⅹ多糖-0.015ⅹ蛋白+330ⅹ粘度-0.004ⅹ异戊醇+0.06ⅹ乙酸乙酯。
4、结果分析
基于SRC绝对值筛选大于0.3的物质组分,包括选自乳酸、琥珀酸、苹果酸、柠檬酸、丙酮酸的有机酸,选自Ca2+、Mg2+、K+、Al3+的阳离子,选自SO4 2-、PO4 3-、Cl-的阴离子以及涩感多酚、多糖、蛋白和粘度。
上述步骤S2中,对上述筛选得到的组分进行复筛,具体为:
1、样品配制:按照啤酒浓度的10倍复配不同组分的单溶液,包括乳酸、琥珀酸、苹果酸、柠檬酸、丙酮酸、CaCl2、MgCl2、KCl、AlCl3、CaSO4、NaH2PO4、麦芽糊精(多糖)、大豆蛋白(蛋白)、多酚(阿魏酸/没食子酸/咖啡酸/单宁酸)、增稠剂(粘度)。
2、将配制溶液与唾液进行沉淀反应:
依据沉淀指数是否>2%将可能性啤酒组分分为与唾液发生沉淀反应的组分和与唾液无沉淀反应的组分。
进一步对上述两类组分进行分析,具体为:
对与唾液发生沉淀反应的组分进行粒径分析,将粒径>500nm的组分初判为涩感因素(与唾液发生沉淀反应带来涩感的物质);在一优选实施例中,所述涩感因素包括乳酸、琥珀酸、苹果酸、柠檬酸、CaCl2、CaSO4和涩感多酚(没食子酸/单宁酸)。
对与唾液无沉淀反应的组分进行粘度分析,将粘度增加的组分初判为掩涩因素(降低摩擦系数、提高润滑性能,可以掩盖涩感的物质);在一优选实施例中,所述掩涩因素包括多糖和蛋白,其中所述多糖包括淀粉多糖和非淀粉多糖。
上述步骤S3中,对上述得到的涩感因素和掩涩因素进行涩感强度判断,具体为:
对于涩感因素:
分别以啤酒浓度的50%反添加入啤酒中,根据沉淀指数增加幅度以及粒径大小判断涩感因素的最终影响程度具体为:
对于粒径500-1000nm的组分,其沉淀指数的增加幅度<10%,定义为III类;>10%,定义为II类;
对于粒径1000-2000nm的组分,其沉淀指数的增加幅度<5%,定义为III类;5%-10%,定义为II类;>10%,定义为I类;
对于粒径>2000nm的组分,其沉淀指数的增加幅度<5%,定义为II类;>5%,定义为I类;
其中,I类为影响程度大;II类为影响程度中等;III类为影响程度小。
在一优选实施例中,涩感负面因素的最终影响程度具体为:
对于掩涩因素:
向啤酒中以万分之一体积比,外添加不同的专一性酶制剂,降解属于掩涩因素的各组分,然后测试粘度及摩擦系数。粘度降低越大,摩擦系数增加越高,代表该组分的掩涩效果越好。对不同组分排序,得出掩涩因素作用强度。具体为:
掩涩因素的最终影响程度为非淀粉多糖、淀粉多糖和蛋白。
基于上述分析,最终确定了啤酒涩感因素为涩感多酚(I类)、苹果酸、柠檬酸和乳酸(II类)以及琥珀酸、硫酸钙和氯化钙(III类);掩涩因素为非淀粉多糖、淀粉多糖以及蛋白。
为了验证影响因素,进行多轮次工艺试验,目的在于对于涩感非常突出的某品类啤酒,如何通过降低涩感因素和/或提高掩涩因素来降低其涩感,提高滑口性,并通过感官品评判定改变影响因素后是否有效。
实施例
感官品评方法分析
1)品评小组:固定人员7-8人(包含国家级品酒师4人以上),饮食清淡,下午14:00-17:30之间品评;
2)样品准备:样品浓度一致,所有样品恒温15度;
3)涩感感官描述:涩感关注舌面摩擦感和饮后舌面摩擦感;
4)涩感品评流程:大口饮入啤酒样品液体在口腔中流动10s以上,感受液体在口腔流动过程中以及吞咽后舌面摩擦感强度;样品和样品之间清水漱口,样品之间停留1min;
5)涩感得分:≤1:无涩感或者涩感较轻;1-3轻微涩感;≥3,涩感明显。分值越高,代表涩感越强。具体结果如表2所示。
表2工艺调整前后涩感影响因素(涩感因素和掩涩因素)含量变化以及感官品评结果
基于表2数据发现,试验1测试了降低了II类涩感因素-乳酸含量,涩感明显改善;试验2测试了降低III类涩感因素硫酸钙,涩感降低,但幅度不如乳酸降低明显;试验3取消了酶制剂添加,提高了掩涩因素,涩感也得到了有效改善。试验4降低涩感因素同时提高了掩涩因素,涩感改善效果最为显著,样品已经无涩感。这表明,本发明提供的涩感因素判定方法与实际工艺试验结果是吻合的。
基于以上分析可见,本发明提供的方法具有以下优势:1)传统涩感改善措施大多针对洗糟,但往往有时改善不显著,这源于涩感影响物质不清晰,主因不明确;本发明系统的阐明了啤酒中涩感的影响因素,既包括带来涩感的物质(涩感因素),还包括掩盖涩感的物质(掩涩因素),有利于明确啤酒涩感的原因以及调控方向;2)本发明提供的影响因素判定方法,采用多种手段更加客观的明确各物质影响强度,比如涩感物质除了与沉淀物含量有关,还与粒径大小有关系,因此更加客观。
Claims (8)
1.涩感物质影响程度的判断方法,其特征在于,包括以下步骤:
利用偏最小二乘回归法建立啤酒唾液蛋白沉淀指数和啤酒8大类物质组分之间的回归方程,筛选回归方程中标准化系数绝对值大于0.3的组分,以该组分作为影响啤酒涩感的可能性啤酒组分;
将上述筛选得到的可能性啤酒组分按照啤酒浓度的10倍分别配制单物质溶液,通过分别与唾液进行沉淀反应,依据沉淀指数是否>2%将可能性啤酒组分分为与唾液发生沉淀反应的组分和与唾液无沉淀反应的组分,对与唾液发生沉淀反应的组分进行粒径分析,将粒径>500nm的组分初判为涩感因素;对与唾液无沉淀反应的组分进行粘度分析,将粘度增加的组分初判为掩涩因素;
根据不同组分与唾液反应的沉淀指数的增加幅度以及粒径大小判断属于涩感因素的各组分的最终影响程度,以及根据粘度及摩擦系数的变化程度判断属于掩涩因素的各组分的最终影响程度。
2.根据权利要求1所述的判断方法,其特征在于,所述8大类物质包括有机酸、阴离子和阳离子、多糖、蛋白、多酚及涩感多酚、醇酯、苦味酸和粘度。
3.根据权利要求1所述的判断方法,其特征在于,利用偏最小二乘回归法建立啤酒唾液蛋白沉淀指数和啤酒8大类物质组分之间的回归方程具体为:
Y=-370+0.078ⅹ乳酸-1.52ⅹ琥珀酸/苹果酸+0.709ⅹ柠檬酸+0.726ⅹ草酸+0.377ⅹ丙酮酸-0.02ⅹ富马酸-0.04ⅹ乙酸+0.032ⅹ总有机酸+0.004ⅹK++229ⅹMn2++0.106ⅹNa++34.7ⅹBa2+-0.686ⅹCa2+-104ⅹAl3++0.378ⅹMg2+-0.0711ⅹ总阳离子-0.0018ⅹCl--0.0008ⅹPO4 3--0.3ⅹSO4 2-+0.056ⅹ涩感多酚+0.0028ⅹ总醇-0.02ⅹ总酯+0.103ⅹ苦味酸-0.22ⅹ多糖-0.015ⅹ蛋白+330ⅹ粘度-0.004ⅹ异戊醇+0.06ⅹ乙酸乙酯。
4.根据权利要求1所述的判断方法,其特征在于,标准化系数绝对值大于0.3的组分包括选自乳酸、琥珀酸、苹果酸、柠檬酸、丙酮酸的有机酸,选自Ca2+、Mg2+、K+、Al3+的阳离子,选自SO4 2-、PO4 3-、Cl-的阴离子以及涩感多酚、多糖、蛋白和粘度。
5.根据权利要求1所述的判断方法,其特征在于,所述涩感因素包括乳酸、琥珀酸、苹果酸、柠檬酸、Ca2+、SO4 2-和涩感多酚;所述掩涩因素包括多糖和蛋白,其中所述多糖包括淀粉多糖和非淀粉多糖。
6.根据权利要求5所述的判断方法,其特征在于,将得到的涩感因素分别以啤酒浓度的50%反添加入啤酒中,根据沉淀指数增加幅度以及粒径大小判断涩感因素的最终影响程度具体为:
对于粒径500-1000nm的组分,其沉淀指数的增加幅度<10%,定义为III类;>10%,定义为II类;
对于粒径1000-2000nm的组分,其沉淀指数的增加幅度<5%,定义为III类;5%-10%,定义为II类;>10%,定义为I类;
对于粒径>2000nm的组分,其沉淀指数的增加幅度<5%,定义为II类;>5%,定义为I类;
其中,I类为影响程度大;II类为影响程度中等;III类为影响程度小。
8.根据权利要求5所述的判断方法,其特征在于,向啤酒中以万分之一体积比外添加不同的专一性酶制剂分别降解属于掩涩因素的各组分,其中,对应淀粉多糖加入的是葡糖淀粉酶,对应非淀粉多糖加入的是葡聚糖酶和木聚糖酶的复合物,对应蛋白加入的是蛋白酶;
掩涩因素的最终影响程度为非淀粉多糖、淀粉多糖和蛋白。
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