CN109283025A - 一种珍珠粉中所含多肽种类的检测方法 - Google Patents
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Abstract
本发明涉及一种珍珠粉中所含多肽种类的检测方法,它包括以下步骤:(a)将珍珠粉加入水中,在不断搅拌的条件下加热至沸并保温得水解液;(b)向所述水解液中加入固体硫酸铵,进行搅拌盐析得盐析物;(c)将所述盐析物溶于NaCl溶液中,随后加入层析柱中,使用所述NaCl溶液进行洗脱得洗脱液;(d)使用紫外分光光度计对所述洗脱液进行检测获得多肽的特征峰数量;(e)采用正丁醇‑醋酸作为展开剂对所述洗脱液进行提取分离,使用茚三酮显色确定斑点数量;当所述斑点数量与所述特征峰数量一致时即可。从而可以证明和解释珍珠的抗衰老的药理作用。
Description
技术领域
本发明属于珍珠粉领域,涉及一种检测方法,具体涉及一种珍珠粉中所含多肽种类的检测方法。
背景技术
珍珠是一种传统名贵中药,在我国已有二千多年的历史,具有养颜、美容、抗衰老、安神、生肌等多方面功效。珍珠所含成份综多,不同类成份所起作用也不同,药理研究证明多肽具有抗衰老作用,因此需要对珍珠中多肽类成份的数量进行确定和分离,以验证珍珠的抗衰老作用。
发明内容
本发明目的是为了克服现有技术的不足而提供一种珍珠粉中所含多肽种类的检测方法。
为达到上述目的,本发明所采用的技术方案为:一种珍珠粉中所含多肽种类的检测方法,它包括以下步骤:
(a)将珍珠粉加入水中,在不断搅拌的条件下加热至沸并保温得水解液;
(b)向所述水解液中加入固体硫酸铵,进行搅拌盐析得盐析物;
(c)将所述盐析物溶于NaCl溶液中,随后加入层析柱中,使用所述NaCl溶液进行洗脱得洗脱液;
(d)使用紫外分光光度计对所述洗脱液进行检测获得多肽的特征峰数量;
(e)采用正丁醇-醋酸作为展开剂对所述洗脱液进行提取分离,使用茚三酮显色确定斑点数量;当所述斑点数量与所述特征峰数量一致时即可。
优化地,它还包括:
(f)以0.1M的醋酸溶液为缓冲液对所述洗脱液进行电泳检测,使用茚三酮显色以再次核对斑点数量。
优化地,步骤(e)中,所述斑点的数量为两个,其Rf轴数值分别为0.35、0.43。
优化地,步骤(a)中,所述珍珠粉与所述热水的质量比为1:1~3,所述保温时间为1~3h。
优化地,步骤(b)中,所述固体硫酸铵与所述水的比例为10~20g:100ml。
优化地,步骤(c)中,所述NaCl溶液的浓度为0.5~1.0g/ml。
由于上述技术方案运用,本发明与现有技术相比具有下列优点:本发明珍珠粉中所含多肽种类的检测方法,通过对珍珠粉进行水解、盐析等化学处理,并结合展开分离、紫外分光光度计的检测实现了对多肽种类的核定和分离,从而可以证明和解释珍珠的抗衰老的药理作用。
附图说明
图1为本发明中紫外分光光度计检测获得的图谱示意图。
具体实施方式
本发明珍珠粉中所含多肽种类的检测方法,它包括以下步骤:(a)将珍珠粉加入水中,在不断搅拌的条件下加热至沸并保温得水解液;(b)向所述水解液中加入固体硫酸铵,进行搅拌盐析得盐析物;(c)将所述盐析物溶于NaCl溶液中,随后加入层析柱中,使用所述NaCl溶液进行洗脱得洗脱液;(d)使用紫外分光光度计对所述洗脱液进行检测获得多肽的特征峰数量;(e)采用正丁醇-醋酸作为展开剂对所述洗脱液进行提取分离,使用茚三酮显色确定斑点数量;当所述斑点数量与所述特征峰数量一致时即可。通过对珍珠粉进行水解、盐析等化学处理,并结合展开分离、紫外分光光度计的检测实现了对多肽种类的核定和分离,从而可以证明和解释珍珠的抗衰老的药理作用。
上述检测方法还包括以下步骤:(f)以0.1M的醋酸溶液为缓冲液对所述洗脱液进行电泳检测,使用茚三酮显色以再次核对斑点数量,这样能够进一步确定多肽的种类,以保证检测的准确程度。步骤(e)中,所述斑点的数量为两个,其Rf轴数值分别为0.35、0.43。步骤(a)中,所述珍珠粉与所述热水的质量比为1:1~3,所述保温时间为1~3h。步骤(b)中,所述固体硫酸铵与所述水的比例为10~20g:100ml。步骤(c)中,所述NaCl溶液的浓度为0.5~1.0g/ml。
下面将结合实施例对本发明进行进一步说明。
实施例1
本实施例提供一种珍珠粉中所含多肽种类的检测方法,它包括以下步骤:
(a)将珍珠粉加入水中(珍珠粉与水的质量比为1:2),在不断搅拌的条件下加热至沸(约100℃)并保温2h得水解液;
(b)向水解液中加入固体硫酸铵(硫酸铵与水的比例为15g:100ml),进行搅拌盐析得盐析物;
(c)将盐析物溶于NaCl溶液(NaCl溶液的浓度为0.9g/ml,盐析物和NaCl溶液的比例约为20g:100ml)中,随后加入层析柱中,使用相同浓度的NaCl溶液进行洗脱得洗脱液;
(d)使用紫外分光光度计对洗脱液进行检测获得多肽的特征峰数量(如图1所示);
(e)采用正丁醇-醋酸(体积比为1:1)作为展开剂对洗脱液进行提取分离,使用茚三酮显色确定斑点数量(斑点数量也为两个,其Rf轴数值分别为0.35、0.43);当斑点数量与特征峰数量一致时即可(当斑点数量与特征峰数量不一致时,还可以继续以下步骤);
(f)以0.1M的醋酸溶液为缓冲液对所述洗脱液进行电泳检测,使用茚三酮显色以再次核对斑点数量(最终确定珍珠粉中多肽的种类为两种)。
实施例2
本实施例提供一种珍珠粉中所含多肽种类的检测方法,它与实施例1中的基本一致,不同的是:步骤(a)中,珍珠粉与水的质量比为1:1,保温1h得水解液;步骤(b)中,向水解液中加入固体硫酸铵,使得硫酸铵与水的比例为10g:100ml;步骤(c)中,NaCl溶液的浓度为0.5g/ml。
实施例3
本实施例提供一种珍珠粉中所含多肽种类的检测方法,它与实施例1中的基本一致,不同的是:步骤(a)中,珍珠粉与水的质量比为1:3,保温3h得水解液;步骤(b)中,向水解液中加入固体硫酸铵,使得硫酸铵与水的比例为20g:100ml;步骤(c)中,NaCl溶液的浓度为1g/ml。
上述实施例只为说明本发明的技术构思及特点,其目的在于让熟悉此项技术的人士能够了解本发明的内容并据以实施,并不能以此限制本发明的保护范围。凡根据本发明精神实质所作的等效变化或修饰,都应涵盖在本发明的保护范围之内。
Claims (6)
1.一种珍珠粉中所含多肽种类的检测方法,其特征在于,它包括以下步骤:
(a)将珍珠粉加入水中,在不断搅拌的条件下加热至沸并保温得水解液;
(b)向所述水解液中加入固体硫酸铵,进行搅拌盐析得盐析物;
(c)将所述盐析物溶于NaCl溶液中,随后加入层析柱中,使用所述NaCl溶液进行洗脱得洗脱液;
(d)使用紫外分光光度计对所述洗脱液进行检测获得多肽的特征峰数量;
(e)采用正丁醇-醋酸作为展开剂对所述洗脱液进行提取分离,使用茚三酮显色确定斑点数量;当所述斑点数量与所述特征峰数量一致时即可。
2.根据权利要求1所述珍珠粉中所含多肽种类的检测方法,其特征在于,它还包括:
(f)以0.1M的醋酸溶液为缓冲液对所述洗脱液进行电泳检测,使用茚三酮显色以再次核对斑点数量。
3.根据权利要求1所述珍珠粉中所含多肽种类的检测方法,其特征在于,步骤(e)中,所述斑点的数量为两个,其Rf轴数值分别为0.35、0.43。
4.根据权利要求1所述珍珠粉中所含多肽种类的检测方法,其特征在于:步骤(a)中,所述珍珠粉与所述热水的质量比为1:1~3,所述保温时间为1~3h。
5.根据权利要求1所述珍珠粉中所含多肽种类的检测方法,其特征在于:步骤(b)中,所述固体硫酸铵与所述水的比例为10~20g:100ml。
6.根据权利要求1所述珍珠粉中所含多肽种类的检测方法,其特征在于:步骤(c)中,所述NaCl溶液的浓度为0.5~1.0g/ml。
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