CN108802160B - 一种采用毛细管电泳-质谱联用分析鉴定落叶松中二氢槲皮素的方法 - Google Patents
一种采用毛细管电泳-质谱联用分析鉴定落叶松中二氢槲皮素的方法 Download PDFInfo
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Abstract
本发明提供了一种使用毛细管电泳‑质谱联用(CE‑MS)法检测落叶松中二氢槲皮素的方法。所述方法包括步骤为:(1)制备纯品二氢槲皮素的标准溶液,在毛细管电泳仪中进样,在质谱仪中分析,获得质谱图;(2)绘制标准曲线和计算最低检测限;(3)采用步骤(1)的方法获得样品的质谱图,计算样品加样回收率;(4)根据样品色谱峰与特征子离子信息,对落叶松中二氢槲皮素进行鉴定;(5)根据标准曲线对落叶松中二氢槲皮素进行定量分析。本发明具有操作简单、节约时间、检测高效、重复性好等特点,所需样品用量少,检测结果准确可靠,是一种对落叶松中二氢槲皮素进行定性、定量分析的优良方法。
Description
技术领域
本发明涉及二氢槲皮素的鉴定方法,具体为采用毛细管电泳-质谱联用(CE-MS)建立的二氢槲皮素的分析鉴定方法。
背景技术
二氢槲皮素(Dihydroquercetin,DHQ),又称花旗松素(Taxifolin)、黄杉素、紫杉叶素,是二氢黄酮类化合物的一种,其结构式如下。在自然界中,二氢槲皮素的含量较低,但分布却较为广泛,如广泛存在于松科、金柳梅科、蔷薇科、漆树科、唇形科等植物中,其中在落叶松的根颈部含量最高。落叶松被离体后,其二氢槲皮素的含量随储存时间的增长而不断降低。
二氢槲皮素最初是由日本学者Fukui提取出来的一种葡萄糖苷元,分子式为C15H12O7,分子量为304.25,是无色针尖状结晶(50%乙醇)或淡黄色粉末,熔点240℃,无特殊气味,几乎不能溶于苯等非极性溶剂,能稍溶于水,且易溶于极性溶剂(如乙醇、乙酸)、沸水中。
通常采用薄层扫描、分光光度计法和液相色谱法分析二氢槲皮素的含量,但薄层扫描和分光光度计法检测结果的准确性相对较低,导致两种方法在进行准确的定量分析中应用性不高。高效液相色谱法是现有检测方法中最为有效的方法,但依旧存在着分析成本高、样品使用量大、日常仪器维护费用贵,检测时间较长的缺点。
快速、精确、灵敏的新型鉴定方法仍为市场及科研所需要。毛细管电泳-质谱联用作为一种新兴的检测技术,利用了毛细管电泳具有的精密度高,样品用量小,分离速度快,单针样品测量费用低等特点,还具有质谱的定性能力强,能够给出分子局部的结构信息等特点,是多数检测标准中采用的仪器设备。将这一技术应用到二氢槲皮素的分析中,可以完善现有的检测鉴定方法。
发明内容
针对目前二氢槲皮素鉴定方法中分析成本高、样品使用量大,检测时间长的缺点,本发明提供了一种采用毛细管电泳-质谱联用的二氢槲皮素鉴定方法,能够高效、快速的鉴定落叶松中的二氢槲皮素。
为了实现上述目的,本发明采用了如下技术方案:
一种使用毛细管电泳-质谱联用法检测落叶松中二氢槲皮素的方法,所述方法包括的步骤为:
(1)制备纯品二氢槲皮素的标准溶液,在毛细管电泳仪中进样,获得所述纯品二氢槲皮素的质谱图;其中所述纯品二氢槲皮素以甲醇为溶剂,配制梯度浓度的二氢槲皮素标准溶液;所述在毛细管电泳中进样为20mbar进样2s,15-30kV条件下进行电泳,紫外波长选择214nm,柱温为20-35℃,缓冲盐为乙酸铵溶液;质谱条件为阳离子模式,MS/MS全扫描,氮气流速15psi,氦气7psi,气体温度为300℃,分离电压3.0kV,鞘流液使用50%的甲醇溶液(含0.01%醋酸铵),流量为100nL/min,泵流速为0.4L/min,分流比为100:1;
(2)根据步骤(1)中质谱图的峰面积进行积分计算,绘制标准曲线和计算最低检测限;
(3)将样品二氢槲皮素和一份纯品二氢槲皮素按照步骤(1)的参数在毛细管的电泳仪中进样,在质谱仪中分析,获得质谱图,计算加样回收率;其中所述样品二氢槲皮素包括配制好的落叶松植物样品一份,添加本步骤所述纯品二氢槲皮素的落叶松样品一份;根据得到所述色谱图计算色谱峰面积,采用步骤(1)中标准曲线公式计算所述色谱峰面积所对应的进样量,分别记为纯品二氢槲皮素S0、植物样品S1、添加纯品二氢槲皮素的植物样品S2,回收率公式为(2S2-S1)/S0×100%;
(4)根据步骤(3)中色谱峰与特征子离子信息,对所述落叶松中二氢槲皮素进行鉴定;
(5)根据所述标准曲线对所述落叶松中二氢槲皮素进行定量分析。
进一步的,所述步骤(1)中纯品二氢槲皮素的梯度浓度为2000、1000、500、250、125μg/mL,采用Chemstation软件对峰面积进行积分计算。
所述步骤(1)中毛细管电泳的电压为20kV,柱温为25℃,乙酸铵浓度为20mmol。
所述步骤(3)中所述纯品二氢槲皮素的浓度为0-2mg/mL。
所述步骤(3)中所述纯品二氢槲皮素的浓度为125μg/mL。
本发明所述方法较现有技术具有以下突出的优点和显著的技术效果:
1、本方法首次将毛细管电泳联用质谱仪用于标准品二氢槲皮素以及落叶松中天然二氢槲皮素的检测,并建立了一种以该仪器为主的高效快速的分析鉴定方法,可用于快速高效检测落叶松提取物保健食品中的二氢槲皮素含量。
2、本方法用时短,测试一针的样品只需要15分钟即能得到结果,液相色谱检测检测的时间约为30-40分钟,节约了一半时间;样品使用量小,只需要几纳升样品即可以进行检测,液相则需要10微升以上,从微升级提升到了纳升级;精确性高,实验结果表明毛细管电泳联用质谱仪对二氢槲皮素的最低检测限为15.6pmol(47588pg),优于液相色谱;实验重复性较高,通过计算RSD都在1%左右,误差值低,可信程度比较高。
3、本方法对二氢槲皮素的裂解进行了研究,利用了质谱联用后能打多级质谱的特点,对二氢槲皮素的子离子进行了详细分析,通过对子离子的分析,不仅能够进一步验证样品是否为二氢槲皮素,还为今后该仪器对生物类黄酮的检测研究提供了思路。
附图说明:
图1为实施例中二氢槲皮素标准品的标准曲线
图2为二氢槲皮素标准品(A)和落叶松木屑植物样品(B)的色谱,紫外光谱,质谱图
图3为二氢槲皮素的裂解行为
具体实施方式
下面以具体实施方式对发明作进一步详细说明
本发明参照特定的实施例进行了描述,但是,很显然仍可以做出各种修改和变换而不违背本发明的精神和范围。因此,本发明的说明书和附图应该认为是说明性的而非限制性的。
下列实施例中,采用的落叶松来自大兴安岭林场,木屑为落叶松根颈部分,其部位二氢槲皮素含量较高。
具体实施例:
(1)以甲醇作溶剂,分别配制2000、1000、500、250、125μg/mL浓度的二氢槲皮素标准溶液。
(2)在毛细管电泳仪中进样,采用20mbar进样2s,在20kV条件下进行电泳,紫外波长选择214nm,柱温为25℃,缓冲盐为20mmol乙酸铵溶液;质谱条件为阳离子模式,MS/MS全扫描,氮气流速15psi,氦气7psi,气体温度为300℃,分离电压3.0kV,鞘流液使用50%的甲醇溶液(含0.01%醋酸铵),流量为100nL/min,泵流速为0.4L/min,分流比为100:1,每个浓度重复上样3次,用Chemstation软件对峰面积进行积分计算,根据得到的结果绘制标准曲线,结果见图1,并测得最低检测限为15.6pmol。
(3)从落叶松中提取二氢槲皮素,将晒干的落叶松木屑放于103-105℃下烘干12小时,然后用粉碎机粉碎至细,用80目筛过筛;将筛后的原料再放于103-105℃条件下干燥12小时。将过筛后进行干燥的落叶松称取30mg于试管中,用2mL 60%乙醇提取,先在4℃条件下存放48h,超声30min,再用乙酸乙酯与丙酮进行萃取,用盐酸调pH为3,收集有机相,过0.45μm膜,烘干后得二氢槲皮素,使用时用甲醇溶解,定容至2mL,将落叶松样品进行毛细管电泳-质谱分析并与标准品进行比对,结果见图2。
(4)计算加样回收率,取稀释1/8的二氢槲皮素标准品一份(125μg/mL),配制好的落叶松植物样品一份,再取添加稀释1/8的二氢槲皮素标准品的落叶松样品一份进行检测,测量后计算出色谱峰面积,用标准曲线公式计算这些峰面积所对应的进样量;分别记为标准品S0、植物样品S1、添加标准品的植物样品S2,使用回收率公式(2S2-S1)/S0×100%进行加样回收率的测定,回收率为93.25%符合要求。
(5)按照得到的质谱图对样品进行裂解结果分析(图3,表1),进一步确认检测结果为二氢槲皮素。
(6)根据标准曲线对落叶松中二氢槲皮素进行定量分析,计算得落叶松中二氢槲皮素得率为0.52%。
表1二氢槲皮素的子离子碎片信息
Claims (5)
1.一种使用毛细管电泳-质谱联用法检测落叶松中二氢槲皮素的方法,其特征为,所述方法包括的步骤为:
(1)制备纯品二氢槲皮素的标准溶液,在毛细管电泳仪中进样,在质谱仪中分析,获得所述纯品二氢槲皮素的质谱图;其中所述纯品二氢槲皮素以甲醇为溶剂,配制梯度浓度的二氢槲皮素标准溶液;所述在毛细管电泳中进样为20mbar进样2s,15-30kV条件下进行电泳,紫外波长选择214nm,柱温为20-35℃,缓冲盐为乙酸铵溶液;质谱条件为阳离子模式,MS/MS全扫描,氮气流速15psi,氦气7psi,气体温度为300℃,分离电压3.0kV,鞘流液使用含0.01%醋酸铵的50%的甲醇溶液,流量为100nL/min,泵流速为0.4L/min,分流比为100:1;
(2)根据步骤(1)中质谱图的峰面积进行积分计算,绘制标准曲线和计算最低检测限;
(3)将样品二氢槲皮素和一份纯品二氢槲皮素按照步骤(1)的参数在毛细管电泳仪中进样,在质谱仪中分析,获得质谱图,并计算加样回收率:其中所述样品二氢槲皮素包括配制好的落叶松植物样品一份,和添加本步骤所述纯品二氢槲皮素的落叶松样品一份,根据得到的所述质谱图计算质谱峰面积,采用步骤(2)中标准曲线公式计算所述质谱峰面积所对应的进样量,分别记为纯品二氢槲皮素S0、植物样品S1、添加纯品二氢槲皮素的植物样品S2,回收率公式为(2S2-S1)/S0×100%;
(4)根据步骤(3)中质谱图中质谱峰与特征子离子信息,对所述落叶松中二氢槲皮素进行鉴定,其中所述特征子离子信息为母离子质荷比为305、子离子荷质比为153和258;
(5)根据所述标准曲线对所述落叶松中二氢槲皮素进行定量分析。
2.根据权利要求1所述的方法,所述步骤(1)中纯品二氢槲皮素的梯度浓度为2000、1000、500、250、125μg/mL,采用Chemstation软件对峰面积进行积分计算。
3.根据权利要求1所述的方法,所述步骤(1)中毛细管电泳的电压为20kV,柱温为25℃,乙酸铵浓度为20mmol。
4.根据权利要求1所述的方法,所述步骤(3)中所述纯品二氢槲皮素的浓度为0-2mg/mL。
5.根据权利要求4所述的方法,所述步骤(3)中所述纯品二氢槲皮素的浓度为125μg/mL。
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毛细管电泳紫外检测法测定莲须中的槲皮素、木犀草素、山萘酚、异槲皮甙;荆瑞俊 等;《分析化学研究简报》;20070831;第35卷(第8期);第1187-1190页 |
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