甜瓜保鲜剂的高效液相色谱测定方法的研究
摘要
由于农药品种和用量的不断增加，农药作为对环境和食品的重要污染源之一，越来越受到各国政府和公众的关注。人们不断研究更新、更快捷准确、实用的对农残进行检测的分析方法，特别是多残留分析方法。
本文通过对几种甜瓜保鲜剂的提取、分离、测定和残留动态研究，提出了利用高效液相色谱及液相色谱-质谱联用同时测定几种保鲜剂的方法，这些方法适用于甜瓜中保鲜剂在保鲜过程中及水果贮运中的残留动态分析，也适用于日常生产、生活中瓜果蔬菜等中的保鲜剂残留量测定。
本论文共分五个部分：
1．概述了农药残留分析的一些进展，介绍了常用的农药残留分析样品前处理技术和检测技术以及近期发展的样品前处理新技术和残留量检测新技术，阐述了新疆瓜果保鲜剂残留量测定方法的研究意义。
2．分别论述了从农药原药中提取保鲜剂多菌灵(MBC)、甲基托布津(TFM)、噻菌灵、TBZ)、抑霉唑(IMZ)、扑海因(ID)的方法。对从农药原药中提取标准品的方法进行了研究，针对不同农药剂型及待提取保鲜剂的性质，采用不同的提取方法，制得的标准品均经熔点测定、分子量测定、元素分析等分析手段验证。
3．提出了一种用高效液相色谱法(HPLC)同时测定甜瓜样品中的抑霉唑(IMZ)、噻菌灵(TBZ)含量的方法。用乙酸乙酯作提取剂提取甜瓜样品，通过一个净化步骤分离了样品中共提取组分。采用InertsilC8-3色谱柱(150×4.6mm，5μm)，流动相
为乙睛：水。75：25和用外标法定量，方法的最低检测限分别
为 TBZ：lug，IMZ 2 ng；回收率为 87．5－98刀％，变异系数为
1．5～4．2％。本法用于甜瓜样品中IMZ、TBZ含量的测定，获
得了满意的结果。
4．提出了用高效液相色谱法同时测定甜瓜样品中的多菌灵
（MBC）、甲基托布津（TFM）、噬菌灵（TBZ）含量的方法，
该方法用甲醇作提取剂，用甲醇冰－二氯甲烷体系一次净化完
成，二极管阵列检测器检测，用外标法定量，最低检测限分别
为 MBC：lug，TBZ：lug，TFM：5 ng，变异系数为 0．8～3．2％，
回收率为80．9～97．3％。本法用于甜瓜在保鲜过程中及贮运中
MBC、TBZ、TFM残留量的测定，取得了满意的结果。
5．提出了一种用液相色谱一质谱法同时测定甜瓜样品中的噬菌灵
（TBZ）、甲基托布津（TFM）、扑海因（ID）含量的方法。该
法用溶剂直接提取进样，方法简单、快速；测定采用电喷雾（ESI）
作离子源，检测模式为正离子，定量检测方式为选择离子检测
（SIM），检测离子In／Z：202．25、343卫8、330刀7。最低检测限
分别为＊＊Z：o．1＊，＊＊M：10＊g，ID：3＊；回收率为81．0～
110％；相对标准偏差为1．5～16、8％。方法灵敏度高，选择性好，
准确度高，最低检测限达到pg级。用于甜瓜样品中噬菌灵、甲
基托布津、扑海因的测定，获得了满意的结果。
Pesticides, as one of the major pollutant sources of environment and food, now are becoming more concerned by publics and many countries in the world with the increases of varieties and usage of pesticides. In recent years, new analytical methods, especially multiresidues analysis methods, have been developed greatly. In this paper, through the dynamic study of the pesticide multiresidues in muskmelon, the methods are presented for their simultaneous determination by high-performance liquid chromatograph (HPLC) and liquid chromatography-mass spectrometry (LC-MS). These methods are available to be applied to the dynamic analysis of the pesticides involved in muskmelon during storage and transportation and also to the pesticide residues in vegetable and fruit in regulatory production and daily life. This paper consists of five chapters.
1. A review concerning the analytical methods of the pesticide residues is presented. The regulatory and recent developed technologies of the extraction and instrumental analysis are discussed. The significance of the research in this paper is also given.
2. The procedures purifying pesticide products for analysis standards , i.e., thiabendazole(TBZ), thiophanate methyl(TFM), iprodione(ID), carbendazim(MBC), imazalil(IMZ), are reported. These methods are presented according to the variety of the market pesticide products and the properties of the pesticides mentioned above. The purities of the
references extracted are verified by melting points, molecular mass and elementary analysis.
3. A HPLC method for the simultaneous determination of the amounts of IMZ and TBZ in muskmelons is investigated. The samples were extracted with ethyl acetate followed by a simple clean-up procedure to separate the analytes from the coextractives. Liquid chromatographic separation was performed on an InertsilC8-3 column with acetonitrile-water (75:25) as mobile phase. The method was quantified by an external standard mode. The detection limits of the method were 1 ng for TBZ and 2 ng for IMZ, and the coefficients of variation, recoveries were 1.5-4.2%, 87.5-98.0%, respectively. The method has been applied to the quantitative determination of IMZ and TBZ in muskmelons with satisfactory results. The method is simple, rapid, and accurate.
4. A HPLC method is investigated for simultaneous determination of carbendazim (MBC), thiabendazole (TBZ) and thiophanate-methyl (TFM) in muskmelons. Analyses were performed with simply clean-up procedure followed by HPLC-DAD(diode array detector) detection on a reversed-phase column eluted with acetonitrile- 8 mM phosphate buffer (35:65, v/v). The recoveries of MBC, TBZ and TFM from muskmelon samples were in the range of 80.9-86.4%, 90.8-96.0% and 92.5-97.3% respectively with coefficients of variation ranging from 0.8% to 3.2%. The method was applied to determining MBC, TBZ and TFM in the muskmelons during storage, and the results were satisfied.
5. A HPLC-MS method is presented for the simultaneous determination of the micro amounts of TBZ, TFM, ID in muskmelon samples by HPLC-MS. The method involved a simple and rapid extraction procedure and liquid chromatography coupled to electrospray mass selective detection. lonization was performed at atmospheric pressure in an electrospray-type source (+) and quantitative detection was carried out using the selected ion monitoring (SIM) mode. Limits of detection were 0.1 pg for TBZ, 10 pg for TFM, 3 ng for ID. The average recoveries and relative standard deviations ranged from 81.0-110% and from 1.5-16.8%, respectively. The method was applied to the determination of TBZ, TFM and ID in melon samples with satisfactory results.
本文通过对几种甜瓜保鲜剂的提取、分离、测定和残留动态研究，提出了利用高效液相色谱及液相色谱-质谱联用同时测定几种保鲜剂的方法，这些方法适用于甜瓜中保鲜剂在保鲜过程中及水果贮运中的残留动态分析，也适用于日常生产、生活中瓜果蔬菜等中的保鲜剂残留量测定。
本论文共分五个部分：
1．概述了农药残留分析的一些进展，介绍了常用的农药残留分析样品前处理技术和检测技术以及近期发展的样品前处理新技术和残留量检测新技术，阐述了新疆瓜果保鲜剂残留量测定方法的研究意义。
2．分别论述了从农药原药中提取保鲜剂多菌灵(MBC)、甲基托布津(TFM)、噻菌灵、TBZ)、抑霉唑(IMZ)、扑海因(ID)的方法。对从农药原药中提取标准品的方法进行了研究，针对不同农药剂型及待提取保鲜剂的性质，采用不同的提取方法，制得的标准品均经熔点测定、分子量测定、元素分析等分析手段验证。
3．提出了一种用高效液相色谱法(HPLC)同时测定甜瓜样品中的抑霉唑(IMZ)、噻菌灵(TBZ)含量的方法。用乙酸乙酯作提取剂提取甜瓜样品，通过一个净化步骤分离了样品中共提取组分。采用InertsilC8-3色谱柱(150×4.6mm，5μm)，流动相
为乙睛：水。75：25和用外标法定量，方法的最低检测限分别
为 TBZ：lug，IMZ 2 ng；回收率为 87．5－98刀％，变异系数为
1．5～4．2％。本法用于甜瓜样品中IMZ、TBZ含量的测定，获
得了满意的结果。
4．提出了用高效液相色谱法同时测定甜瓜样品中的多菌灵
（MBC）、甲基托布津（TFM）、噬菌灵（TBZ）含量的方法，
该方法用甲醇作提取剂，用甲醇冰－二氯甲烷体系一次净化完
成，二极管阵列检测器检测，用外标法定量，最低检测限分别
为 MBC：lug，TBZ：lug，TFM：5 ng，变异系数为 0．8～3．2％，
回收率为80．9～97．3％。本法用于甜瓜在保鲜过程中及贮运中
MBC、TBZ、TFM残留量的测定，取得了满意的结果。
5．提出了一种用液相色谱一质谱法同时测定甜瓜样品中的噬菌灵
（TBZ）、甲基托布津（TFM）、扑海因（ID）含量的方法。该
法用溶剂直接提取进样，方法简单、快速；测定采用电喷雾（ESI）
作离子源，检测模式为正离子，定量检测方式为选择离子检测
（SIM），检测离子In／Z：202．25、343卫8、330刀7。最低检测限
分别为＊＊Z：o．1＊，＊＊M：10＊g，ID：3＊；回收率为81．0～
110％；相对标准偏差为1．5～16、8％。方法灵敏度高，选择性好，
准确度高，最低检测限达到pg级。用于甜瓜样品中噬菌灵、甲
基托布津、扑海因的测定，获得了满意的结果。
Pesticides, as one of the major pollutant sources of environment and food, now are becoming more concerned by publics and many countries in the world with the increases of varieties and usage of pesticides. In recent years, new analytical methods, especially multiresidues analysis methods, have been developed greatly. In this paper, through the dynamic study of the pesticide multiresidues in muskmelon, the methods are presented for their simultaneous determination by high-performance liquid chromatograph (HPLC) and liquid chromatography-mass spectrometry (LC-MS). These methods are available to be applied to the dynamic analysis of the pesticides involved in muskmelon during storage and transportation and also to the pesticide residues in vegetable and fruit in regulatory production and daily life. This paper consists of five chapters.
1. A review concerning the analytical methods of the pesticide residues is presented. The regulatory and recent developed technologies of the extraction and instrumental analysis are discussed. The significance of the research in this paper is also given.
2. The procedures purifying pesticide products for analysis standards , i.e., thiabendazole(TBZ), thiophanate methyl(TFM), iprodione(ID), carbendazim(MBC), imazalil(IMZ), are reported. These methods are presented according to the variety of the market pesticide products and the properties of the pesticides mentioned above. The purities of the
references extracted are verified by melting points, molecular mass and elementary analysis.
3. A HPLC method for the simultaneous determination of the amounts of IMZ and TBZ in muskmelons is investigated. The samples were extracted with ethyl acetate followed by a simple clean-up procedure to separate the analytes from the coextractives. Liquid chromatographic separation was performed on an InertsilC8-3 column with acetonitrile-water (75:25) as mobile phase. The method was quantified by an external standard mode. The detection limits of the method were 1 ng for TBZ and 2 ng for IMZ, and the coefficients of variation, recoveries were 1.5-4.2%, 87.5-98.0%, respectively. The method has been applied to the quantitative determination of IMZ and TBZ in muskmelons with satisfactory results. The method is simple, rapid, and accurate.
4. A HPLC method is investigated for simultaneous determination of carbendazim (MBC), thiabendazole (TBZ) and thiophanate-methyl (TFM) in muskmelons. Analyses were performed with simply clean-up procedure followed by HPLC-DAD(diode array detector) detection on a reversed-phase column eluted with acetonitrile- 8 mM phosphate buffer (35:65, v/v). The recoveries of MBC, TBZ and TFM from muskmelon samples were in the range of 80.9-86.4%, 90.8-96.0% and 92.5-97.3% respectively with coefficients of variation ranging from 0.8% to 3.2%. The method was applied to determining MBC, TBZ and TFM in the muskmelons during storage, and the results were satisfied.
5. A HPLC-MS method is presented for the simultaneous determination of the micro amounts of TBZ, TFM, ID in muskmelon samples by HPLC-MS. The method involved a simple and rapid extraction procedure and liquid chromatography coupled to electrospray mass selective detection. lonization was performed at atmospheric pressure in an electrospray-type source (+) and quantitative detection was carried out using the selected ion monitoring (SIM) mode. Limits of detection were 0.1 pg for TBZ, 10 pg for TFM, 3 ng for ID. The average recoveries and relative standard deviations ranged from 81.0-110% and from 1.5-16.8%, respectively. The method was applied to the determination of TBZ, TFM and ID in melon samples with satisfactory results.
引文
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