CN106979988B - 一种蛋白质含量的检测方法 - Google Patents
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Abstract
本发明的目的是提供一种利用氨基酸分析检测蛋白质的方法。包括以下具体步骤:(1)称取样品于水解管中,加入乙醇溶液的对酚磺酸水解试剂,超声使样品混合均匀,通氩气或氮气除去溶解氧,封口,放入真空干燥箱中水解;(2)取出试管水解管冷却至室温,开管后,过滤到50ml容量瓶中,洗涤并收集洗涤液,定容至刻度线;(3)吸取0.5‑2ml定容后的样品于试管中,放入真空干燥箱中使其完全干燥;(4)在蒸干后的样品水解物试管中加入样品缓冲液,置振荡混合器上混合均匀,用针管吸取少量,通过过滤器过滤后,用氨基酸分析仪对水解液进行氨基酸分析,将得到样品的氨基酸图谱与标准样品的谱图对照,根据图谱上各峰的保留时间及峰面积求出样品中的氨基酸种类及含量。(5)在求得的氨基酸质量的基础上数减去与氨基酸总摩尔数相同摩尔数的水分子质量,即得到蛋白质的质量,根据样品的称重量及稀释倍数计算样品中蛋白质的含量。
Description
所属技术领域:
本发明涉及的是一种蛋白质的检测方法,属于食品、饲料及生化领域。
背景技术:
蛋白质的含量是人类食品和动物饲料的主要评价指标,蛋白质也是生物样品的重要组成成分。蛋白质的测定方法很多,主要有凯氏定氮法、双缩脲法、Folin-酚试剂法、紫外吸收法、考马斯亮蓝法等。″食品中蛋白质的测定国家标准″(GB 5009.5-2010)中包含了凯氏定氮法、燃烧法和分光光度法等三种方法。凯氏定氮法和燃烧法测定的是总氮含量,因此得到的结果代表的是粗蛋白含量。这一检测方法无法区分蛋白氮和非蛋白氮。″乳品三聚氰胺事件″就是不法分子利用了凯氏定氮法的这一缺陷,将含氮量高达66.67%的有毒化学品三聚氰胺人为地加入到乳品中,使蛋白质的检测值虚高而牟取暴利。上述其它方法也不能准确有效地对蛋白质含量进行检测。目前,传统的蛋白质检测方法均存在着一定的缺陷,而现代的仪器分析方法尚不成熟。
蛋白质是氨基酸通过肽键结合而成的高分子化合物。食品中的蛋白质的营养价值取决于其构成的氨基酸的种类和比例,按蛋白质中必需氨基酸的含量和种类可将食物中的蛋白质分为三类:(1)完全蛋白:蛋白质组成中含有充足的维持生命和促进生长的全部必需氨基酸,如乳中的酪蛋白、蛋中的卵蛋白,这类蛋白作为食物对人体的营养价值最高;(2)不完全蛋白:蛋白质组成中缺乏一种或几种人体必需氨基酸,它不能维持生命,也不能促进机体生长,如玉米中的玉米胶蛋白、动物结缔组织、肉皮中的胶质蛋白;(3)半完全蛋白:蛋白质中氨基酸组成介于上述两者之间,即蛋白质中含有人体所需的必需氨基酸,但氨基酸间含量比例不平衡,它能维持生命,但不能促进生长,如小麦和大麦中的麦胶蛋白。
将蛋白质水解成氨基酸,再对氨基酸进行分析,这样不仅能准确测定蛋白质的含量,而且可以了解构成蛋白质的氨基酸的种类和比例,并且能更好地评价蛋白质的质量。但是目前报导的蛋白质水解方法,都不能使蛋白质水解产物中的氨基酸真实地反映构成蛋白质的氨基酸,虽然在理论上可行,但是由于水解方法的限制,目前还无法通过该方法准确地测定蛋白质的含量。
文献中报道的常用于蛋白质水解的方法有三种,即酸水解法、碱水解法和酶水解法。酸水解法通常采用的是硫酸(4mol/L)或HCL(6mol/L),回流煮沸20h。其优点是水解完全,不引起消旋作用;缺点为:色氨酸被破坏,羟基氨基酸(丝氨酸及苏氨酸)有一小部分被分解,天冬酰胺和谷胺酰胺的酰胺基被水解下来。另外,也可采用蛋白质水解管,在60℃、无氧条件下水解20小时,该法在同样也存在氨基酸损失的问题。碱水解法是使样品与NaOH(5mol/L)共煮10-20h,优点为:水解完全,色氨酸不受破坏;缺点为:氨基酸产生消旋现象,还会引起精氨酸脱氨并生成鸟氨酸和尿素。酶水解法是用胰蛋白酶,糜蛋白酶,胃蛋白酶等蛋白酶进行分步水解。优点为:氨基酸不产生消旋作用,也不破坏氨基酸;缺点为:采用单种酶往往不能水解完全,需要几种酶协同作用才能使蛋白质完全水解,所需时间较长,步骤复杂,不适合在蛋白质检测中的应用。因此,对采用述三种蛋白质水解方法得到的水解产物进行氨基酸分析,得到的结果都不能真实地反映构成蛋白质的氨基酸种类及比例。
发明内容:
本发明的目的是提供一种蛋白质水解为氨基酸并通过氨基酸分析检测蛋白质含量的方法。
本发明的技术方案是:
首先将样品中的蛋白质进行水解,然后用氨基酸分析仪对水解液中的氨基酸进行分析,根据氨基酸求出蛋白质的含量。
具体实施方案:
1、蛋白质的水解
(1)样品的蛋白质的水解:称取样品于蛋白质水解管中,加入对酚磺酸水解试剂,对酚磺酸水解试剂是以乙醇、甲醇或乙腈的水溶液配制,超声,使样品混合均匀,通氩气或氮气除去溶解氧,封口后,放入真空干燥箱中,于100-120℃水解14-20小时;
(2)过滤及定容:水解完成后,取出水解管冷却至室温,开管后,将水解物过滤,洗涤水解管和滤纸,收集洗涤液,并定容;
(3)蒸干处理:吸取定容后的样品并放入真空干燥箱中,在温度为40-60℃下使其完全干燥,底部留有少许固体或痕渍。
2、氨基酸的测定
(1)氨基酸的分析原理:氨基酸分析仪中的阳离子交换柱利用氨基酸的酸碱性、极性及相对分子质量的不同,使各种氨基酸在色谱柱上实现分离。氨基酸被缓冲溶液洗脱下来的先后顺序依次是酸性氨基酸、极性大的氨基酸和非极性氨基酸、碱性氨基酸。被洗脱下来的氨基酸与茚三酮发生显色反应,颜色的深浅程度在一定范围内与氨基酸的含量呈线性关系,然后利用分光光度法对氨基酸进行检测,每种被洗脱下来的氨基酸会对应不同的保留时间,根据得到图谱的保留时间可以对氨基酸进行定性分析,根据每个谱峰的面积或峰高可以对其进行定量分析。
(2)氨基酸标准样品的分析
氨基酸标准品1:标准品中含有19种氨基酸,分别为半胱氨酸(Cys)、天冬氨酸(Asp)、蛋氨酸砜(MetSON)、苏氨酸(Thr)、丝氨酸(Ser)、谷氨酸(Glu)、脯氨酸(Pro)、甘氨酸(Gly)、丙氨酸(Ala)、胱氨酸((Cys)2)、缬氨酸(Val)、蛋氨酸(Met)、异亮氨酸(Ile)、亮氨酸(Leu)、酪氨酸(Tyr)、苯丙氨酸(Phe)、组氨酸(His)、赖氨酸(Lys)和精氨酸(Arg),标准品中每种氨基酸浓度为100nmol/mL,标准品1的图谱如图1所示。
氨基酸标准品2:色氨酸(Trp)的浓度为100nmol/mL。标准品2的图谱如图2所示。
采用德国aminoSys公司的A300氨基酸分析仪,进样量为20μL,每种氨基酸参与显色反应的物质的量为2nmol。
(3)样品的氨基酸分析及蛋白质含量的计算
在蒸干后的样品水解物试管中加入样品缓冲液,置振荡混合器上混合均匀,用针管吸取少量,通过过滤器过滤后,用A300型氨基酸分析仪对水解液进行氨基酸分析,进样量为20μL,得到样品的氨基酸图谱。与标准样品的谱图对照,根据图谱上各峰的保留时间及峰面积求出样品中的氨基酸种类及含量。
根据样品峰和标准品峰面积之比,求算水解液中每种氨基酸摩尔数,公式为:
式中:ni-样品中氨基酸的摩尔数,mol;
si-样品中每种氨基酸的峰面积;
s0-标准品氨基酸的峰面积;
2-标准品进样中的氨基酸纳摩尔数,nmol;
0.02-进样量,ml;
50-样品的稀释倍数。
样品中每种氨基酸的质量为:
mi=ni×Mi (2)
式中:mi-每种氨基酸的质量,g;
Mi-氨基酸的相对分子质量。
蛋白质的水解相当于每两个氨基酸之间加入了一个水分子,由于蛋白质属于高分子化合物,分子量非常大,每个蛋白质由成千上万个氨基酸构成,因此,计算时只按氨基酸的摩尔数减去相同摩尔数的水分子求算蛋白质,而由于每个蛋白质链之间相差的1个水分子产生的误差可以忽略不计。样品中蛋白质的百分含量为:
式中:mi-每种氨基酸的质量,g;
ni-样品中氨基酸的摩尔数,mol;
m-样品的称样量,g。
本发明具有如下优点:
1、本发明中采用的蛋白质水解剂为对酚磺酸的,较传统的盐酸或氢氧化钠水解剂温和,对于色氨酸不产生破坏作用;
2、对酚磺酸水解剂是以乙醇、甲醇或乙腈的水溶液配制,降低了溶液的极性,较传统的水溶剂极性低,对水解产生的氨基酸起到了保护作用;
3、因为作为食品中蛋白质的营养价值主要是由必需氨基酸决定的,因此,采用氨基酸分析来检测蛋白质,可以了解构成蛋白质的氨基酸的种类及各自的含量,更有利于评价食品的营养价值。
实例1
1、样品的蛋白质水解:称取奶粉样品0.2000g于水解管中,样品是由唐山市产品质量监督检验所提供的,是牛乳企业由鲜牛乳直接干燥得到的,在水解管中加入5ml浓度为6mol/L的对酚磺酸水解试剂,对酚磺酸水解试剂是以20%乙醇溶液配制的,超声30min使样品混合均匀,通氩气或氮气除去溶解氧,封口后,放入真空干燥箱中,于110℃水解17h;水解完成后取出水解管冷却至室温,开管后,过滤到50mL容量瓶中,用水洗涤水解管和滤纸,收集洗涤液,并定容至刻度线;吸取定容后的样品1mL放入真空干燥箱中,在温度为50℃下使其完全干燥,底部留有少许固体或痕渍。
2、氨基酸的分析:在蒸干后的样品水解物试管中加入1mL样品缓冲液,置振荡混合器上混合均匀,用针管吸取少量,通过过滤器过滤后,用A300型氨基酸分析仪对水解液进行氨基酸分析,进样量为20μL,得到样品的氨基酸图谱3。与氨基酸标准样品的谱图1和谱图2进行对照,根据图谱上各峰的保留时间及峰面积求出样品中的氨基酸种类及含量,见表1。
表1对酚磺酸水解干奶粉的氨基酸分析数据
求出氨基酸的总摩尔数为416.4,氨基酸的总质量为54990求得样品中蛋白质的百分含量为23.74%。
3、对比实验:称取相同的奶粉样品0.2000g于水解管中,将用5ml浓度为6mol/L的盐酸为水解试剂进行水解,在温度为110℃时水解样品24h,其它步骤与1和2的相同,进行氨基酸分析得到谱图,如图4所示。根据图谱上各峰的保留时间及峰面积求出样品中的氨基酸种类及含量,见表2。
表2盐酸水解干奶粉的氨基酸分析数据
求出氨基酸的总摩尔数为300.2μmol,氨基酸的总质量为39340μg,计算得出样品中蛋白质的百分含量为16.97%。盐酸作为水解剂与对酚磺酸比较,前者的水解液中不含有色氨酸及半胱氨酸,而后者均能检测到色氨酸及半胱氨酸,说明盐酸作为水解试剂使色氨酸和半胱氨酸完全破坏。另外,盐酸作为水解剂求得的蛋白质含量是16.97%,而以对酚磺酸为水解试剂求得的蛋白质的含量是23.74%,后者得到的蛋白质含量及各种氨基酸的含量均明显高于前者,说明了盐酸作为水解剂会使氨基酸受到明显破坏。
实例2
1、样品的蛋白质水解:称取市售商品奶粉样品0.2000g于水解管中,在水解管内加入6ml浓度为5.5mol/L的对酚磺酸水解试剂,对酚磺酸水解试剂是以20%乙腈溶液配制,超声25min使样品混合均匀,通氩气或氮气除去溶解氧,封口后,放入真空干燥箱中,于105℃水解20h;水解完成后取出水解管冷却至室温,开管后,过滤到50mL容量瓶中,用水洗涤水解管和滤纸,收集洗涤液,并定容至刻度线;吸取定容后的样品1mL放入真空干燥箱中,在温度为55℃下使其完全干燥,底部留有少许固体或痕渍。
2、氨基酸的分析:在蒸干后的样品水解物试管中加入1mL样品缓冲液,置振荡混合器上混合均匀,用针管吸取少量,通过过滤器过滤后,用A300型氨基酸分析仪对水解液进行氨基酸分析,进样量为20μL,得到样品的氨基酸图谱5。
3、计算样品中蛋白质的含量:将图5与氨基酸标准品的谱图1和2进行比较,根据图谱上各峰的保留时间及峰面积求出样品中的氨基酸种类及含量,见表3
表3对酚磺酸水解市售奶粉的氨基酸分析数据
求出氨基酸的总摩尔数为375.7μmol,氨基酸的总质量为49160μg,计算求得样品中蛋白质的百分含量为21.20%。样品中的色氨酸含量较高,说明采用该水解方法可以很好地保护氨基酸不受破坏。
附图说明:
图1氨基酸标准品1的图谱。
图2氨基酸标准品2的图谱。
图3对酚磺酸水解干奶粉的氨基酸分析谱图。
图4盐酸水解干奶粉的氨基酸图谱。
图5对酚磺酸水解市售奶粉的氨基酸分析图谱。
Claims (3)
1.一种蛋白质水解为氨基酸并通过氨基酸分析检测蛋白质含量的方法,其特征在于,包括以下步骤:
(1)样品的蛋白质的水解:称取样品于蛋白质水解管中,加入对酚磺酸水解试剂,对酚磺酸水解试剂是以乙醇、甲醇或乙腈的水溶液配制,超声,使样品混合均匀,通氩气或氮气除去溶解氧,封口后,放入真空干燥箱中,于100-120℃水解14-20小时;
(2)过滤及定容:水解完成后,取出水解管冷却至室温,开管后,将水解物过滤,用水洗涤水解管和滤纸,收集洗涤液,并定容;
(3)蒸干处理:吸取定容后的样品并放入真空干燥箱中,在温度为40-60℃下使其完全干燥,底部留有少许固体或痕渍;
(4)氨基酸的测定:得到的水解氨基酸样品用缓冲溶液稀释,采用氨基酸分析仪进行分析,根据样品峰和标准品峰面积之比,求算水解液中每种氨基酸的含量,并计算得到样品中蛋白质的含量。
2.根据权利要求1所述的一种蛋白质水解为氨基酸并通过氨基酸分析检测蛋白质含量的方法,其特征在于,采用对酚磺酸作为蛋白质水解试剂,较传统的盐酸或氢氧化钠水解剂温和,对于色氨酸不产生破坏作用,水解用对酚磺酸的浓度为5.5mol/L或6mol/L。
3.根据权利要求1所述的一种蛋白质水解为氨基酸并通过氨基酸分析检测蛋白质含量的方法,其特征在于,配制对酚磺酸水解剂的试剂是采用乙醇、甲醇或乙腈的水溶液配制,降低了溶液的极性,较传统的水溶剂极性低,对水解产生的氨基酸起到了保护作用,采用20%的乙醇、甲醇或乙腈的水溶液配制对酚磺酸水解剂。
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---|---|---|---|---|
CN104789713A (zh) * | 2015-04-16 | 2015-07-22 | 佛山市顺德区龙亭新材料有限公司 | 含铬合成鞣剂的方法及制革方法 |
CN105586444A (zh) * | 2016-01-06 | 2016-05-18 | 四川大学 | 一种用酚磺酸水解制革废弃物制备的皮革复鞣填充剂 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2685601A (en) * | 1950-04-06 | 1954-08-03 | Sterling Drug Inc | Method of hydrolyzing proteins to amino acids |
US2991309A (en) * | 1957-12-30 | 1961-07-04 | Internat Minerals & Chemicals | Protein hydrolysis |
US3952109A (en) * | 1973-02-13 | 1976-04-20 | Far-Mar-Co., Inc. | Low salt protein hydrolyzates |
FR2596391B1 (fr) * | 1986-03-27 | 1988-12-02 | Flork Sa Laboratoires | Procede de preparation industrielle d'acides amines par hydrolyse de proteines en milieu acide |
US6858114B2 (en) * | 2001-07-03 | 2005-02-22 | The University Of Alabama | Laser hydrolysis of polypeptides |
AU2004231997A1 (en) * | 2003-04-16 | 2004-11-04 | The University Of Mississippi | Immunostimulatory agents in botanicals |
US9179687B2 (en) * | 2005-03-30 | 2015-11-10 | Board Of Trustees Of The University Of Arkansas | Yeast fermentation of rice bran extracts |
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104789713A (zh) * | 2015-04-16 | 2015-07-22 | 佛山市顺德区龙亭新材料有限公司 | 含铬合成鞣剂的方法及制革方法 |
CN105586444A (zh) * | 2016-01-06 | 2016-05-18 | 四川大学 | 一种用酚磺酸水解制革废弃物制备的皮革复鞣填充剂 |
Non-Patent Citations (4)
Title |
---|
Simplified protein hydrolysis with methanesulfonic acid at elevated temperature for complete amino acid analysis of proteins;SHYH-HORNG CHIOU et al;《Journal of Chromatography》;19881231;第448卷;第404-410页 * |
应用磺酸水解法进行人血白蛋白氨基酸组成分析;赵宗建 等;《生物化学与生物物理进展》;19881231;第15卷(第4期);第316、301页 * |
羽毛氨基酸含量测定影响因素分析;林香信 等;《中国农学通报》;20111231;第27卷(第1期);第389-393页 * |
蛋白质水解阶段对氨基酸组成分析的影响;陈泽宪 等;《分析科学学报》;20020228;第18卷(第1期);第80-85页 * |
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