1、中华人民共和国出入境检验检疫行业标准SN/T 1593-2005 进出口蜂蜜中五种有机磷农药残留量检验方法气相色谱法Inspection of five organophosphorus pesticides residues in honey for export and import-Gas chromatography 2005嗣05-20发布2005-12-01实施中华人民共和国发布国家质量监督检验检疫总局前言本标准的附录A为资料性附录。本标准由国家认证认可监督管理委员会提出并归口。本标准起草单位:中华人民共和国浙江出入境检验检疫局。本标准主要起草人:谢文、丁慧瑛、朱晓雨、郑自强、章晓
2、氧、楼成杰。本标准系首次发布的出入境检验检疫行业标准。SN/T 1593-2005 I SN/T 1593-2005 1 范围进出口蜂蜜中五种有机磷农药残留量检验方法气相色谱法本标准规定了蜂蜜中5种有机磷农药残留量检验的抽样、制样和气相色谱检验方法。本标准适用于蜂蜜中敌百虫、皮蝇磷、毒死蝉、马拉硫磷、蝇毒磷农药残留量的检验。2 抽样和制样2. 1 检验批以不超过1000件为一个检验批。同一检验批的商品应具有相同的特征，如包装、标记、产地、规格、等级等。2.2 抽样鼓量抽样数量见表1。表1抽样数量批量最低抽样数50以内5 50100 10 101500 每增加100，增取5501以上每增加100
3、，增取22.3 抽样工具2.3.1 取样管:不锈钢管或有机玻璃管，长约115cm，直径约2.5cm 2.3.2 混昆样器:搪瓷桶(或杯)。2.3.3 样品瓶:500mL磨砂盖广口玻璃瓶。2.4 抽样方法单位为件按2.2规定的抽样件数随机抽样，逐件开启。将取样管缓缓放人，吸取样品，如遇蜂蜜结晶时，则用取样管插到底，抽取样品。每件抽取样品不少于100g，作为原始样品。将所取样品倾入混样器，7昆合均匀，取500g样品装入清洁干燥的样品瓶内，封口后，标明标记并及时送实验室。2.5 试样制备对未结晶的样品将其用力搅拌均匀，对有结晶析出的样品可将样品瓶盖塞紧后，置于不超过60.C的水浴中温热，待样品全部融
4、化后搅匀，迅速冷却至室温。在融化时应注意防止水分挥发。制备好的试样均分成两份，分别装入样品瓶中，密封，并标明标记。2.6 试样保存在抽样和制样的操作过程中，应防止样品污染或发生残留物含量的变化。试样应于室温下保存。3 试验方法3. 1 方法提要蜂蜜加水稀释后，用乙酸乙醋提取样品中有机磷农药，低温浓缩，用配有火焰光度检测器的气相色谱仪测定，外标法定量。SN/T 1593-2005 3.2 试剂和材料除另有规定外，所有试剂均为分析纯，水为蒸馆水。3.2.1 乙酸乙醋为色谱纯。3.2.2 氧化铀。3.2.3 敌百虫标准品:纯度大于97%;皮蝇磷标准品:纯度大于99%;毒死蝉标准品:纯度大于99%;
5、马拉硫磷标准品:纯度大于99%;蝇毒磷标准品z纯度大于99%。3.2.4 敌百虫、皮蝇磷、毒死蝉、马拉硫磷、蝇毒磷标准储备摇液:分别称取O.0100 0 g标准品(3.2.3)，分别用乙酸乙醋溶解定容至100mL，溶液浓度为100g/mL，存放于40C冰箱。临用前根据需要用乙酸乙醋稀释至适当浓度，作为1昆合标准工作液。3.3 仪器和设备3.3. 1 气相色谱仪，配有火焰光度检测器(磷滤光片525nm)。3.3.2 旋转蒸发器。3.3.3 离心机:4000 r/min。3.3.4 71昆匀器。3.4 测定步骤3.4. 1 提取称取10g均匀试样(精确到0.01g)，置于50mL离心管中，加入15
6、mL水和5g氯化销，泪匀，再加入15mL乙酸乙醋，在混匀器上混匀2min，离心5min(3 000 r/mi时，用尖嘴吸管将上层乙酸乙醋溶液移至100mL浓缩瓶中，水相再加入15mL乙酸乙醋，重复上述操作，合并两次提取液，用旋转蒸发器在400C水洛减压榷缩至近干，加入8mL乙酸乙醋榕解，定量转移至10mL离心管中，在400C以下水浴中用平缓氮气流吹至干，准确加入1mL乙酸乙醋，混匀，供气相色谱测定。3.4.2 测定3.4.2. 1 色谱条件进行测定的色谱条件如下:a) 色谱柱:石英毛细管柱，DB-1701，30mXO. 25 mm(内径)X 0.25m(膜厚)，或相当者;b) 载气:氮气(纯度
7、大于99.999%);载气流速1.0 mL/min;尾吹气流速:30mL/min;氢气流速:75 mL/min，空气流速:100 mL/min; c) 柱温:初始温度500C保持2min，以150C/min升至1800C保持1min，以80C/min升至2900C保持10min; d) 进样口温度:2500C ; e) 检测器温度:2500C;f) 进样方式:不分流进样;g) 进样量:2L;h) 开阀时间:1. 5 min。3.4.2.2 色谱测定根据样品中被测有机磷农药的含量，选定峰面积相近的标准工作榕液。标准工作榕液和样液中各种有机磷的响应值均应在仪器的线性范围内。标准工作梅液和样液等体积
8、穿插进样测定。在上述色谱条件下敌百虫的保留时间约为6.0min，皮蝇磷的保留时间约为17.8min，毒死蝉的保留时间约为18.5 min，马拉硫磷的保留时间约为18.9min，蝇毒磷的保留时间约为27.8mino标准品的色谱图参见附录A中图A.103.4.3 空白试验除不加试样外，均按上述操作步骤进行。2 SN/T 1593-2005 3.4.4 结果计算和襄述用色谱数据处理机或按式(1)计算试样中各种有机磷的残留含量，计算结果需扣除空白值:式中zX=AXcXV 一二AsXm X一一试样中各种有机磷残留量，单位为毫克每千克(mg/kg); A一一样液中各种有机磷的峰面积，单位为平方厘米(cm2
9、); As -标准工作液中各种有机磷的峰面积，单位为平方厘米(cm2); C一一标准工作液中各种有机磷的浓度，单位为微克每毫升(g/mL); V一一样液最终定容体积，单位为毫升(mL);m一一称样量，单位为克(g)。4 测定低限和回收率4. 1 测定低限., ( 1 ) 敌百虫0.01mg/kg、皮蝇磷0.01mg/kg、毒死蝉0.01mg/kg、马拉硫磷0.01mg/kg、蝇毒磷0.01 mg/kg。4.2 回收率回收率的实验数据(在不同添加浓度范围内)如下za) 敌百虫z1) 添加浓度在0.01mg/kg回收率为70%85%; 2) 添加浓度在0.02mg/kg回收率为70%90%;D 添
10、加浓度在0.1mg/kg回收率为81%89%。b) 皮蝇磷:1) 添加浓度在0.01mg/kg回收率为73%99%;2) 添加浓度在0.02mg/kg回收率为75%95%;3) 添加浓度在0.1mg/kg回收率为80%97%。c) 毒死蝉:1) 添加浓度在0.01mg/kg回收率为84%102%; 2) 添加浓度在0.02mg/kg回收率为80%100%; 3) 添加浓度在0.1mg/kg回收率为84%106%。d) 马拉硫磷:1) 添加浓度在0.01mg/kg回收率为84%106%; 2) 添加浓度在0.02mg/kg回收率为80%105%; 3) 添加浓度在0.1mg/kg回收率为81%1
11、03%。e) 蝇毒磷:1) 添加浓度在0.01mg/kg回收率为74%96%; 2) 添加浓度在0.02mg/kg回收率为75%100%; 3) 添加浓度在0.1mg/kg回收率为80%102%。3 SNjT 1593-2005 创阳怪附录A(资料性附录)标准晶色谱固辈击目辈5 町、山10 12.5 15 17.5 20 22.5 25 5 7.5 4 图.15种有机磷标准晶的气相色i曾固27.5 mm SN/T 1593-2005 Foreword Annex A of this standard is an informative annex. This standard was prop
12、osed by and is under the charged of certification and accreditation administa tion of the People 5 Republic of China. This standard was drafted by Zhejiang Entry-Exit Inspection and Quarantine Bureau. The standard was mainly drafted by Wen Xie, Hui-yin Ding , Xiao-yu Zhu , Zi-qiang Zhe吨，Xiao-dongZha
13、ng , Cheng-jie Lou This standard is a professional standard for entry-exit inspection and quarantine promulgated for the first time. 5 SN/T 1593-2005 Inspection of five organophosphorus pesticides residues in honey for export and import-Gas chromatography 1 Scope The standard specifies the methods o
14、f sampling , sample preparation and determination by gas chro matography CGC) of five organophosphorus pesticides residues in honey. This standard is applicable to the determination of trichlorfon, fenchlorphos , chlorprifos， malathi on , coumaphos. 2 Sampling and sample preparation 2. 1 Inspection
15、lot 丁hequantity of an inspection lot should not be more than 1 000 packages. The characteristics of the cargo within the same inspection lot, such as packing , mark, origin,speci fication , grade etc. , should be the same. 2. 2 Ouantity of the sample taken Ouantity of the sample taken see Table 1. N
16、umber of packages in each inspection lot Up to 50 50.100 101.500 more than 501 2. 3 Sampling tools Table 1 Minimum number of packages to be taken 5 10 take 5 more packages for every increment of 100 packages take 2 more packages for every increment of 100 packages 2.3.1 Sampling tube: stainless stee
17、l pipe or colored glasses ppe; length: about 115 cm , diame ter: about 2. 5 cm. 2.3.2 Sample mixer: enamel drum(or cup). 2.3.3 Sample bottle: 500 mL wide-mouth glass bottle with ground stopper. 2. 4 Sampleing procedure 6 SN/T 1593-2005 Take the sample b;nsert;ng the sampling tube slowly ;nto the pac
18、kage after the packages were taken at random according to 2.2 and opened one bone. If the honey is crystallized. the single casing tube should be used and penetrated to the bottom of the container. The sample taken from each package shall be not less than 100 9 as primary sample. Pour the samples ta
19、ken into the sample mix 肘.and m;x thoroughly. then take 500 9 sample pour into a clean and dry sample bottle. seal. label. and send to the laboratorin time. 2. 5 Preparation of test sample The sample which is not crystallized shall be stirred well to make homogeneous. If the sample is crystallized.
20、it must be warmed in a water-bath below 60 with the sample bottle covered tightly; mix thoroughly when all sample has melted. then cool immedately to room temperature. In the course of melting the sample. precautions must be taken to avoid evaporation of water from the sample. Keep the prepared samp
21、le into two sample bottles. seal and label. 2. 6 Storage of the sample In the course of sampling and sample preparation. precautions must be taken to avoid contamination or any factors which macause the. change of residue content. The test sample is stored at room temperature. 3 Method of determinat
22、ion 3.1 Principle Honey is diluted by distilled water. then organophosphorus pesticides are extracted from the diluted solution by ethyl acetate. The extracted solution is concentrated under the lower temperture. Finally residues are determined by GC-FPD. using external standard method. 3. 2 Reagent
23、s and materials Unless otherwise speci fied .剖Ithe reagent used should be analytical grade. water is distilled wa ter. 3. 2. 1 Ethyl acetate: HPLC grade 3.2.2 Sodium chloride:analytical grade. 3.2.3 Standard of trichlorfon: purity97%. Standard of fenchlorphos: purty99%. Standard of chlorpyrifos: pur
24、ity99%. Standard of malathion: purity 99% Standard of coumaphos: purity 99%. 3.2.4 Stock standard solution of trichlorfon. fenchlorphos. chlorpyrifos. malathon. coumaphos: accurately weigh 0.01000 9 standard(3. 2.韵.dissolve with ethyl acetate and quantitatively on 100 mL volumetric flask individuall
25、. the concentration of solutions are 100g/mL. should store in 4t refrigerator. According to the concentration required. the mixed standard workng solution is pre-7 SN/T 1593-2005 pared from the stock solution by ethyl acetate before using. 3.3 Apparatus and equipment 3.3.1 Gas chromatograph: equippe
26、d with flame photmetric detector (phosphorous filter). 3. 3. 2 Rotary vacuum evaporator. 3.3.3 Centrifuge: 4000 r/min. 3. 3. 4 Vortex mixer. 3. 4 Procedure 3. 4. 1 Extraction Weigh ca 10 9 of the test sample (accurate to 0.01 g) into a 50 mL centrifuge tube, add 15 rnL wa ter, 5. 0 9 sodium chloride
27、 and mix it, add 15 mL ethyl acetate, blend for 2 min , centrifuge for 5 min under 3 000 r/min. Transfer the upper ethI acetate layer into the flask. Water layer repeat the pro cedure with 15 mL ethyl acetate, combine ethyl acetate extracted solution and evaporate to nearly dryness in a water bath b
28、elow 40oC. Transfer the risudes with 8 mL ethyl acetate into 10 mL centri fuge tube, blow just dryness with N2 gas in a water bath below 4Ot. Add exactl1 mL ethyl acetate to dissolve the residue , mix the solution. The solution is ready for GC determination. 3. 4. 2 Oetermination 3.4.2. 1 GC operati
29、ng conditions a) Column: fused silica capillary column , 08-1701 30 m x 0.25 mm X 0.25m， or the equi回国lent. b) Carrier gas and make up gas: Nitrogen (purit 99. 999%) , flow rate of carrier gas: 1. 0 mL/min, flow rate of make up gas: 30 mL/min, flow rate of hydrogen: 75 mL/min, flow rate of air: 100
30、mL/min. c) Temperature programme: 50t (keep 2 min) 15t /min to 180t (keep 1 min) 8t /min to 2900C Ckeep 10 min). d) Injection temperature: 250oC. e) Oetector temperature: 250oC. f) Injection mode: splitless. g) Injection volume: 2L h) Purge time: 1.5 min. 3.4. 2. 2 GC determination According to the
31、values of five organophosphorus pesticides in sample , select the standard working solution with similar peak area to that of sample solution. The responses of organophosphorus in the standard working and the sample solution should be within the linear range of the instrumental 8 SN/T 1593一2005detec
32、tion. The standard working solution should be randomly injected in between the injections of sample solution of equal volume. Under the above GC operating condition , the retention time of tri chlorfon is about 6.0 min , fenchlorphos is about 17.8 min , chlorpyrifos is about 18.53 min, malathi on is
33、 about 18.9 min , coumaphos is about 27.8 min ,. For chromatogram of the standard , see Figure A. 1 in annex A. 3. 4. 3 Blank test The operation of the blank test is the same as the described in the method of determination , but with the omission of sample addition. 3. 4. 4 Calculation and expressio
34、n of result Calculation the content of multi-organophosphorus residue in the test sample by GC data processor or according to the formula (1) The blank value should be subtracted from the above result of calcula tion. Where: X一生羊L羊v-A , Xm X-the residue content of multi organophosphorus in the test
35、sample , mg/kg; A-the peak area of multi organophosphorus in sample solution , cm勺A.一一一thepeak area of multi organophosphorus in standard working solution , cm勺C一一一theconcentration of multi organophosphorus in working solution，g!mL; V一一-thefinal volume of the sample solution , mL; m一一-Massof test sa
36、mple, g. 4 Limit of determination and recover 4. 1 Limit of determination The limit of determination of this method are: . ( 1 ) trichlorfon O. 01 mg!闸，fenchlorphosO. 01 mg!kg , chlorpyrifos O. 01 mg!1唱，malathionO. 01 mg!闸，coumaphos 0.01 mg!kg. 4.2 Recover According to the experimental data , the co
37、rresponding recoveries of fortifing concentrations are: a) trichlorfon: spike O. 01 mg!闸，therecovery is 70% -85%; spike O. 02 mg!闹，therecovery is 70% -90% ; spike O. 1 mg!kg , the recovery is 81 % -89%. b) fenchlorphos: spike O. 01 mg!闸，therecovery is 73% -99%; spike 0.02 mg!1唱，therecove叩is75%-95%;
38、spike O. 1 mg!1唱，therecove叩is80%-97%. c) chlorprifos: spikeO.01 mg!kg,the recovery is84%-102%; 9 SN/T 1593-2005 10 spi ke O. 02 mg/ kg ,the recovery i s 80 % -100 % ; spike O. 1 mg/kg,the recovery is 84%-106%. d) malathion: spike O. 01 mg/kg,the recove叩is84%-106%; spike O. 02 mg/kg, the recove叩is80%
39、 -105%; spike 0.1 mg/kg,the recovery is 81%-103%. e) coumaphos: spike 0.01 mg/kg,the recove叩is74%-96%; spike 0.02 mg/kg, the recovery is 75% -100%; spike O. 1 mg/阔，therecove叩is80%-102%. 口。=。205Annex A (informative annex) Chromatogram of the standards iii 山10 12.5 15 17.5 20 22.5 25 5 7.5 SN/T 1593-2
40、005 Fig. A. 1 the chromatogram of five organophosphorus pesticides standards 的。samES。J 27.5 mm mCON-仍FFHZm中华人民共和国出入境检验检疫行业标准进出口蜂蜜中五种有机磷农药残留量检验方法气相色谱法SN/T 1593-2005 * 中国标准出版社出版发行北京复兴门外三里河北街16号邮政编码，100045网址电话，685239668517548 中国标准出版社秦皇岛印刷厂印刷特印张l字数20千字2005年8月第一次印刷开本880X12301/16 2005年8月第一版9略定价10.00元如有印装差错由本社发行中心调换版权专有侵权必究举报电话:(010)68533533书号:155066 2-16337 SN/T 1593-2005
