蜂花粉和蜂胶中4种砷化合物形态测定方法的研究
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摘要
砷元素在自然界中以无机和有机两种形态广泛存在，它的毒性不仅与其含量有关，更大程度上取决于它的存在形态，无机砷毒性远大于有机砷，已有研究证明无机砷为致癌物质。砷污染严重危害着人类的健康。近年来砷污染问题在蜂产品中也有报道。蜂产品营养丰富，富含人体所需的营养成分、微量元素及酶类等，有“天然营养素”之称，在国内外已被广泛地开发利用。但其食用安全性，尤其是砷污染问题引起了人们极大的关注。目前砷元素形态分析中，氢化物发生-原子荧光光谱（HG-AFS）以其低灵敏度、成本低、操作简单的特点，与高效液相色谱（HPLC）联用逐渐成为主要的检测手段之一。因此，本文建立了利用高效液相色谱-氢化物发生-原子荧光光谱（HPLC-HG-AFS）联用技术测定蜂花粉和蜂胶中砷形态的分析方法。结果如下：
1、建立了以0.20mol/L硫酸为提取溶剂，高效液相色谱-氢化物发生-原子荧光光谱（HPLC-HG-AFS）联用技术作为检测手段，分析蜂花粉中亚砷酸（AsⅢ）、砷酸（AsV）、一甲基砷酸（MMA）和二甲基砷酸（DMA）的方法。AsⅢ、DMA、MMA和AsV的检测限分别是1.0、2.1、1.2、3.1μg/kg，加标回收率均在79%～100%之间，精密度试验结果（n=4）显示四种砷形态的相对标准偏差分别为4.5%、2.3%、11.0%、7.7%。
2、建立了以1%（V/V）磷酸-甲醇（1:1）为提取溶剂，高效液相色谱-氢化物发生-原子荧光光谱联用技术为检测手段，分析蜂胶中四种砷形态化合物的方法。并比较了1%磷酸-甲醇（1:1）与0.20mol/L硫酸-甲醇（1:1）作提取溶剂的优劣，发现前者方法稳定性好、准确性高。AsⅢ、DMA、MMA和AsV的检测限分别是0.6、1.7、1.2、1.6μg/kg。四种砷形态的平均回收率均在66%～110%之间。精密度试验结果（n=6）显示四种砷的相对标准偏差均小于7%。
蜂产品中砷元素形态分析为建立安全可靠的蜂产品安全及营养评价体系提供了科学技术支持和鉴定的标准。
Arsenic compounds exist widely with inorganic and organic arsenic form in nature. Theactual toxicity levels for different arsenic compounds vary greatly. Inorganic arsenic isconsidered the most acutely toxic form. The inorganic compounds of arsenic have beenclassified as carcinogenic. Arsenic contamination of surroundings caused serious harm to humanhealth. Now Arsenic contamination of bee products was also reported. Bee products were rich innutrition ingredients, trace elements and enzymes that human body needs, which were named as"Natural nutrients," and were diffusely development and utilized both at home and abroad. Whiletheir edible safety, especially the arsenic contamination caused great attention. Hydridegeneration and atomic fluorescence spectrometry(HG-AFS) together with high performanceliquid chromatography (HPLC) would become one of the main means of detection gradually inarsenic speciation analysis field at present, with its high sensitivity, low cost and simpleoperation. So the methods for determination of arsenic speciation in bee pollen and propolis byhigh performance liquid chromatography-hydride generation-atomic fluorescence spectrometrywere established in this study. The results were as follows:
1. A method about the determination of arsenic speciation in bee pollen was established.The arsenic compounds, including arsenite(AsⅢ), monomethylarsonic acid (MMA),dimethylarsinic acid (DMA) and arsenate(AsV), in bee pollen were analyzed with highperformance liquid chromatography-hydride generation-atomic fluorescence spectrometry(HPLC-HG-AFS) as detection means and0.20mol/L sulfuric acid as extraction. The detectionlimits for AsⅢ, DMA, MMA and AsⅤ were1.0、2.1、1.2、3.1μg/kg, respectively, and therecoveries of standard addition were in the range of79%～100%with that the relative standarddeviations (RSD) were4.5%,2.3%,11.0%,7.7%, respectively.
2. A method to detect the four arsenic compounds above mentioned in propolis wasestablished, with high performance liquid chromatography-hydride generation-atomicfluorescence spectrometry (HPLC-HG-AFS) as detection means, and with1%(V/V)phosphoricacid-methnol (1:1) as extraction.1%(V/V)phosphoric acid-methnol (1:1) and0.20mol/L sulfuricacid-methnol (1:1) were compared as extraction. It was found that the former had better stability,higher accuracy. The detection limits for As Ⅲ, DMA, MMA and AsⅤ were0.6、1.7、1.2、1.6 μg/kg, respectively, and the recoveries of standard addition were in the range of66%～110%with the relative standard deviations (RSD) all less than7%.
The arsenic speciation analysis in bee products would provide science and technologysupport and the standard of appraisal for establishing safe and reliable bee products safetysystem and nutrition evaluation system.
1、建立了以0.20mol/L硫酸为提取溶剂，高效液相色谱-氢化物发生-原子荧光光谱（HPLC-HG-AFS）联用技术作为检测手段，分析蜂花粉中亚砷酸（AsⅢ）、砷酸（AsV）、一甲基砷酸（MMA）和二甲基砷酸（DMA）的方法。AsⅢ、DMA、MMA和AsV的检测限分别是1.0、2.1、1.2、3.1μg/kg，加标回收率均在79%～100%之间，精密度试验结果（n=4）显示四种砷形态的相对标准偏差分别为4.5%、2.3%、11.0%、7.7%。
2、建立了以1%（V/V）磷酸-甲醇（1:1）为提取溶剂，高效液相色谱-氢化物发生-原子荧光光谱联用技术为检测手段，分析蜂胶中四种砷形态化合物的方法。并比较了1%磷酸-甲醇（1:1）与0.20mol/L硫酸-甲醇（1:1）作提取溶剂的优劣，发现前者方法稳定性好、准确性高。AsⅢ、DMA、MMA和AsV的检测限分别是0.6、1.7、1.2、1.6μg/kg。四种砷形态的平均回收率均在66%～110%之间。精密度试验结果（n=6）显示四种砷的相对标准偏差均小于7%。
蜂产品中砷元素形态分析为建立安全可靠的蜂产品安全及营养评价体系提供了科学技术支持和鉴定的标准。
Arsenic compounds exist widely with inorganic and organic arsenic form in nature. Theactual toxicity levels for different arsenic compounds vary greatly. Inorganic arsenic isconsidered the most acutely toxic form. The inorganic compounds of arsenic have beenclassified as carcinogenic. Arsenic contamination of surroundings caused serious harm to humanhealth. Now Arsenic contamination of bee products was also reported. Bee products were rich innutrition ingredients, trace elements and enzymes that human body needs, which were named as"Natural nutrients," and were diffusely development and utilized both at home and abroad. Whiletheir edible safety, especially the arsenic contamination caused great attention. Hydridegeneration and atomic fluorescence spectrometry(HG-AFS) together with high performanceliquid chromatography (HPLC) would become one of the main means of detection gradually inarsenic speciation analysis field at present, with its high sensitivity, low cost and simpleoperation. So the methods for determination of arsenic speciation in bee pollen and propolis byhigh performance liquid chromatography-hydride generation-atomic fluorescence spectrometrywere established in this study. The results were as follows:
1. A method about the determination of arsenic speciation in bee pollen was established.The arsenic compounds, including arsenite(AsⅢ), monomethylarsonic acid (MMA),dimethylarsinic acid (DMA) and arsenate(AsV), in bee pollen were analyzed with highperformance liquid chromatography-hydride generation-atomic fluorescence spectrometry(HPLC-HG-AFS) as detection means and0.20mol/L sulfuric acid as extraction. The detectionlimits for AsⅢ, DMA, MMA and AsⅤ were1.0、2.1、1.2、3.1μg/kg, respectively, and therecoveries of standard addition were in the range of79%～100%with that the relative standarddeviations (RSD) were4.5%,2.3%,11.0%,7.7%, respectively.
2. A method to detect the four arsenic compounds above mentioned in propolis wasestablished, with high performance liquid chromatography-hydride generation-atomicfluorescence spectrometry (HPLC-HG-AFS) as detection means, and with1%(V/V)phosphoricacid-methnol (1:1) as extraction.1%(V/V)phosphoric acid-methnol (1:1) and0.20mol/L sulfuricacid-methnol (1:1) were compared as extraction. It was found that the former had better stability,higher accuracy. The detection limits for As Ⅲ, DMA, MMA and AsⅤ were0.6、1.7、1.2、1.6 μg/kg, respectively, and the recoveries of standard addition were in the range of66%～110%with the relative standard deviations (RSD) all less than7%.
The arsenic speciation analysis in bee products would provide science and technologysupport and the standard of appraisal for establishing safe and reliable bee products safetysystem and nutrition evaluation system.
引文
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