CN110373446B - 啤酒霉味预估方法 - Google Patents
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Abstract
本发明提供一种啤酒霉味预估方法,涉及啤酒检测领域。本方法包括以下步骤:过滤:将待检测啤酒进行膜过滤;培养:将上述过滤后的滤膜平铺于培养基上进行培养;观察:观察培养基上是否出现菌落,若未出现菌落,则预判该批次啤酒不会产生霉味;若出现菌落,则进行洗涤步骤;洗涤:用洗液对出现菌落的培养基表面进行洗涤,收集洗涤后的洗涤液;检测:检测收集的洗涤液中2,4,6‑三氯苯甲醚的含量,检测同批次啤酒的顶空氧含量,根据预定的判定标准,以2,4,6‑三氯苯甲醚含量和顶空氧含量预判啤酒是否会产生霉味。本发明的方法,可以将检测时间缩短至1.5~3天,而且预测准确率高。
Description
技术领域
本发明涉及啤酒检测领域,特别是涉及啤酒霉味预估方法。
背景技术
啤酒经长时间贮存后,可能会有少数批次的啤酒会出现发霉味道的现象,严重影响成品风味口感。常规监控啤酒是否发霉的方法为:在保存期内,对留样啤酒进行定期品评,判断其是否会产生霉味。但是,这种方法的检测周期非常长,一旦检测发现有霉味的样品,要对该批次的啤酒进行召回,可能会造成一定的经济损失和声誉损失。
发明内容
基于此,有必要针对现有方法检测啤酒是否会产生霉味的方法周期过长的问题,提供一种预测啤酒是否会产生霉味的方法,可以将检测时间缩短至1.5~3天,而且预测准确率高。
一种啤酒霉味预估方法,包括以下步骤:
过滤:将待检测啤酒进行膜过滤;
培养:将上述过滤后的滤膜平铺于培养基上进行培养;
观察:观察培养基上是否出现菌落,若未出现菌落,则预判该批次啤酒不会产生霉味;若出现菌落,则进行洗涤步骤;
洗涤:用洗液对出现菌落的培养基表面进行洗涤,收集洗涤后的洗涤液;
检测:检测收集的洗涤液中2,4,6-三氯苯甲醚的含量,检测同批次啤酒的顶空氧含量,根据预定的判定标准,以2,4,6-三氯苯甲醚含量和顶空氧含量预判啤酒是否会产生霉味。
上述预测方法,对待检啤酒进行膜过滤,用于富集啤酒中的霉菌和霉菌孢子,而培养基可以为霉菌或霉菌孢子提供大量营养物质,使霉菌快速生长。若待测啤酒中没有霉菌或霉菌孢子,则培养基上不会生长出霉菌,该批次啤酒不会产生霉味。若待测啤酒中含有霉菌或霉菌孢子,霉菌会产生2,4,6-三氯苯甲醚(TCA),而此物质是令啤酒产生霉味的主要物质,因此本发明将2,4,6-三氯苯甲醚含量作为鉴定参数之一;另一方面,霉菌在氧含量较低的情况下,生长受到抑制,可能不会令啤酒产生霉味,只有当初始霉菌含量达到一定数量且啤酒内氧含量到达一定程度才会令啤酒产生可识别的霉味。因此,本发明设置2,4,6-三氯苯甲醚含量和顶空氧含量同时作为鉴定参数,可以准确预估将来啤酒是否会产生霉味。
在其中一个实施例中,所述检测步骤中,判定标准为:若所述洗涤液中2,4,6-三氯苯甲醚的含量大于20ng/L,且顶空氧含量大于300ppb,则预判该批次啤酒会产生霉味;若所述洗涤液中2,4,6-三氯苯甲醚的含量≤20ng/L,和/或顶空氧含量≤300ppb,则预判该批次啤酒不会产生霉味。当两个参数均超过阈值时才说明啤酒会产生霉味。
在其中一个实施例中,所述过滤步骤具体为:在无菌条件下,将待检测啤酒通过0.45~1.00μm的微孔膜进行膜过滤。优选地,微孔膜的孔径为0.45μm。
在其中一个实施例中,所述培养步骤具体为:在无菌条件下,将过滤后的滤膜平铺于培养基上,在25-28℃温度下,有氧培养36-72h。
在其中一个实施例中,所述培养基为:察氏琼脂培养基、麦汁琼脂培养基或马铃薯葡萄糖琼脂培养基。优选地,培养基为察氏琼脂培养基。
在其中一个实施例中,所述洗涤步骤中洗液的用量与所述过滤步骤中待检啤酒的体积比为1:5~15。优选地,所述洗液与所述待检测啤酒的体积比为1:10。
在其中一个实施例中,所述洗涤步骤中所用的洗液为无菌水或生理盐水。
在其中一个实施例中,所述生理盐水的质量浓度为0.65%~1.00%。优选地,生理盐水的质量浓度为1.00%。
与现有技术相比,本发明具有以下有益效果:
本发明预测方法,对待检啤酒进行膜过滤,用于富集啤酒中的霉菌和霉菌孢子,而培养基可以为霉菌或霉菌孢子提供大量营养物质,使霉菌快速生长。若待测啤酒中没有霉菌或霉菌孢子,则培养基上不会生长出霉菌,该批次啤酒不会产生霉味。若待测啤酒中含有霉菌或霉菌孢子,霉菌会产生2,4,6-三氯苯甲醚,而此物质是令啤酒产生霉味的主要物质,因此本发明将2,4,6-三氯苯甲醚含量作为鉴定参数之一;另一方面,霉菌在氧含量较低的情况下,生长受到抑制,可能不会令啤酒产生霉味,只有当初始霉菌含量达到一定数量且啤酒内氧含量到达一定程度才会令啤酒产生可识别的霉味。因此,本发明设置2,4,6-三氯苯甲醚含量和顶空氧含量同时作为鉴定参数,可以准确预估将来啤酒是否会产生霉味。
具体实施方式
为了便于理解本发明,下面将结合较佳的实施例对本发明进行更全面的描述。但是,本发明可以以许多不同的形式来实现,并不限于本文所描述的实施例。相反地,提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。
除非另有定义,本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本文中在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不是旨在于限制本发明。
实施例1
一种啤酒霉味预估方法,包括以下步骤:
(1)过滤:在啤酒正常生产中,按生产批次抽取纯生成品啤酒;在无菌条件下,用0.45μm的微孔膜过滤富集500mL成品啤酒;
(2)培养:在无菌条件下,将步骤(1)中过滤啤酒后的微孔膜平铺于含察氏琼脂培养基的培养皿中,将培养皿放置于26℃下,有氧培养48h;
(3)观察:观察步骤(2)中的培养皿是否出现菌落,若未出现菌落,则可断定该批次啤酒不会产生霉味,若出现菌落,则进行洗涤步骤;
(4)洗涤:将50mL质量浓度为1%的灭菌生理盐水分多次对培养基表面菌落进行洗涤,收集洗涤液;
(5)检测:采用气质联用色谱仪检测洗涤液中2,4,6-三氯苯甲醚(TCA)的含量,若洗涤液中2,4,6-三氯苯甲醚的含量大于20ng/L,且该批次的啤酒顶空氧含量大于300ppb,则预判该批次啤酒会产生霉味,若洗涤液中2,4,6-三氯苯甲醚的含量≤20ng/L,和/或顶空氧含量≤300ppb,则预判该批次啤酒不会产生霉味。
实施例2
一种啤酒霉味预估方法,与实施例1的区别在于,洗涤步骤中的生理盐水换为灭菌水。
实施例3
一种啤酒霉味预估方法,与实施例的区别在于,培养步骤中的培养条件为:25℃下有氧培养72h。
实验例1
采用实施例1的方法对某瓶啤酒进行检测,2,4,6-三氯苯甲醚的含量为55.3ng/L,顶空氧的含量为356ppb,说明该批次啤酒会产生霉味。
实验例2
将活化后的标准黑曲霉菌株的按不同浓度接种于灭菌成品啤酒中,灭菌成品啤酒在无菌条件下打开瓶盖,在不同的时间段进行封盖操作,进行此操作是因为正常啤酒中没有霉菌或霉菌含量极少,霉菌含量少的啤酒通常需要很长时间才会产生霉味,在啤酒中接种标准黑曲霉菌株,会加快产生霉味。
本实验中一共有24个样品,每个样品做三个平行样。
采用实施例1的方法对以上24个样品进行检测,与此同时,每种样品留下3瓶于6个月后进行品评,品评是否有霉味。检测结果如下表所示,其中0#为经巴氏灭菌处理的纯生酒液(空白对照),1#~24#为样品:
以上24个样品中,仅24#样品满足以下条件:2,4,6-三氯苯甲醚的含量大于20ng/L,而且顶空氧含量大于300ppb,采用本发明的预估方法24#样品会产生霉味,1#~23#不会产生霉味,而实际风味品评的结果与本发明的结果完全一致。
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (6)
1.一种啤酒霉味预估方法,其特征在于,包括以下步骤:
过滤:在无菌条件下,将待检测啤酒通过0.45~1.00μm的微孔膜进行膜过滤;
培养:在无菌条件下,将过滤后的滤膜平铺于培养基上,在25-28℃温度下,有氧培养36-72h;
观察:观察培养基上是否出现菌落,若未出现菌落,则预判该批次啤酒不会产生霉味;若出现菌落,则进行洗涤步骤;
洗涤:用洗液对出现菌落的培养基表面进行洗涤,收集洗涤后的洗涤液;所述洗液为无菌水或生理盐水;
检测:检测收集的洗涤液中2,4,6-三氯苯甲醚的含量,检测同批次啤酒的顶空氧含量,根据预定的判定标准,以2,4,6-三氯苯甲醚含量和顶空氧含量预判啤酒是否会产生霉味;所述判定标准为:若所述洗涤液中2,4,6-三氯苯甲醚的含量大于20ng/L,且顶空氧含量大于300ppb,则预判该批次啤酒会产生霉味;若所述洗涤液中2,4,6-三氯苯甲醚的含量≤20ng/L,和/或顶空氧含量≤300ppb,则预判该批次啤酒不会产生霉味。
2.根据权利要求1所述的啤酒霉味预估方法,其特征在于,所述培养基为:察氏琼脂培养基、麦汁琼脂培养基或马铃薯葡萄糖琼脂培养基。
3.根据权利要求1~2任一项所述的啤酒霉味预估方法,其特征在于,所述洗涤步骤中洗液的用量与所述过滤步骤中待检啤酒的体积比为1:5~15。
4.根据权利要求3所述的啤酒霉味预估方法,其特征在于,所述洗液与所述待检测啤酒的体积比为1:10。
5.根据权利要求1所述的啤酒霉味预估方法,其特征在于,所述生理盐水的质量浓度为0.65%~1.00%。
6.根据权利要求5所述的啤酒霉味预估方法,其特征在于,所述生理盐水的质量浓度为1.00%。
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