1、中华人民共和国出入境检验检疫行业标准SN/T 1977-2007 进出口水果和蔬菜中睡瞒自旨残留量检测方法高效液相色谱法Determination of fenpyroximate residues in fruit and vegetable for import and export-HPLC method 2007-08-06发布2008-03-01实施战二号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。本标准由中华人民共和国安徽出入境检验检疫局负责起草。本标准主要起草人s郑屏、盛旋、胡艳云、张营、孙吴、朱梦栩。本
2、标准系首次发布的出入挠检验检疫行业标准。SN/T 1977-2007 1 范围进出口水果和蔬菜中瞠蜻醋残留量检测方法高效液相色谱法本标准规定了水果、蔬菜中瞠瞒自旨残留量的高效液相色i昔检测方法。本标准适用于柑桔、白菜中瞠蜗酣残留量的检测。2 方法提要SN/T 1977-2007 试样用甲醉提取，正己统反萃取，中位氧化铝柱净化，以带紫外检测器的高效液相色谱仪检测，外标法定量。3 试剂和材料除特别说明外，所有试剂均为分析纯，水为二次蒸饱水。3. 1 甲那色i普纯。3.2 正己烧z色i普纯。3.3 乙酸乙黯z色i苦纯。3.4 元水硫酸纳，650C灼烧4h，在干燥器中冷却至室1日，贮于密封瓶中备用。3
3、.5 氯化纳溶液，10%。3.6 瞠蜗酣标准品(Fenpyroximate，CAS.NO. ，111812-58-9)，纯度大于等于99%.3.7 瞠瞒自旨标准储备液z准确称取适量盹瞒附标准品(3.6)，用乙脯配制成浓度为100g/mL的标准储备液。根据需要用乙脐稀释至适当浓度的标准工作液，3.8 中性氧化铝固相萃取柱，1000 mg , 3 mL。4 仪器和设备4. 1 高效液相色谱仪，配有紫外检测器。4.2 均质器，5000 r/min。4.3 离心机，4000r/min。4.4 旋转蒸发仪。4.5 氮吹仪。4.6 涡旋混匀器.5 试样制备与保存5. 1 试样制备取代表性样品约500g，取
4、其可食部分，个体大的样品需先切碎，放人高速组织捣碎机捣碎均匀，装人洁净容器，密封，并标明标记。5.2 试样保存试样于0C4C保存。在制样的操作过程中，应防止样品污染或发生残留物含量的变化.1 SN/T 1977-2007 6 测定步骤6. 1 提取称取15g (精确至O.01 g)试样置于50mL离心管中，加入20mL甲醉，高速均质2mino 4 000 r/min离心4mn，过滤，残渣中加入20mL甲醇，涡旋1min，重复离心过滤操作，合并滤液，待净化a6.2 净化将k述滤液转移至250mL分液漏斗中，加入40mL氯化纳溶液(3日和20mL正己烧，剧烈振荡2 min，静置分层，将正己烧层移入
5、100mL具塞锥形瓶中.水层用20mL正己炕重复提取一次，合并正己皖层。加入2g元水硫酸纳，振荡，静置后，将正己统层移人100mL浓缩瓶中，再用少量正己烧洗涤具塞锥形瓶两次，洗涤液合并至浓缩岛;二吃了.-.jt水时此刊队相.x.JLL下。牛二气川斗，听二2用3X3mL正己统+乙酸乙Ii(t千8洗涤浓缩瓶，将溶液转移儿经15mL乙酸乙自由预淋洗的中性氧化铝因相萃取柱。.8)中，控制流diilZUJn;收集全部流出液:i140c水浴氮气吹于，用流动相溶解并定容至1.0 mL.过O.45mil按常被村由苦功斤。.乙旬，中6.3 测定.# 刊于E川油6.3. 1 高效液相色诺条件- a) 色i的主，Z
6、ORAXSB-CI8，5m，j:.-:. bl 流动相z乙脐十水(85+15l，(c贝d) e) 6.4 空白试验除不加试样外，均按上述操作TT7 结果计算和表述用色谱数据处理软件或按式(J) 除空白值) l ( . . . . . . . . . . . . . . . . . . . . . . 式中zx一一试样中瞠蜗E号的残留景，单位为毫克每千克(mg/kgl，A 样液中II:蜗醋的峰面积;As一标准工作液中瞠瞒自旨的峰面积$c一一标准工作液中瞠蹒酷的浓度，单位为微克每毫升(g/mLl; v一一样液最终定容体积，单位为毫升(mLl; m一一政终样液所代表的试样质量，单位为克(g)。B 测
7、定低限和回收率8. 1 测定低限本方法的测定低限为0.02mg/kg. 2 8.2 回收率8.2. 1 柑桔中瞠瞒酣添加浓度及回收率实验数据.一一略蜗醋添加浓度在0.02mg/kg时，回收率为80.4%-97.5%，一一瞠蜗醋添加浓度在0.05mg/kg时，回收率为83.5%-97.1%，一一瞠蜗醋添加浓度在0.20mg/同时，回收率为87.8%-101.7%。8.2.2 白菜中略蹒l添加浓度及回收率实验数据z瞠蹒醋添加浓度在0.02mg/kg时，回收率为78.2%-96.0%，一一瞠蹒醋添加浓度在0.05mg/kg时，回收率为81.7%-97. 4% , 瞠蜗固旨添加浓度在0.20mg/kg
8、时，回收率为87.3%-102.5%。SN/T 1977-2007 3 四月1977-20074 你娟缸 : :.1: ,. 附录A(资料性附录)瞠蹒醋标准晶液相色i昔图2昌恤. 自t l.:I 图A.1瞌蜗醋标准品波相色谙图a咀, SNjT 1977-2007 Foreword Annex A of this standard is an informathe annex. This standard was proposed by and is under the charged of The Certification and Accreditation Ad ministration
9、of the Peoples Republic of China This standard was drafted by Anhui Entry-Exit Inspection and Quarantine Bureau of P. R. C. The standard was mainly drafted by Ping Zheng , Xuan Sheng , Yan-Yun Hu , lei Zhang. Hao Sun. Meng-Xu Zhu. The standard is a professional standard for entry-exit inspection and
10、 quarantine promulgated for the first time. 5 SN/T 1977-2007 Determination of fenpyroximate residues in fruit and vegetable for import and export-HPLC method 1 Scope This standard specifies the methods of sample preparation and determination by high performance liquid chromatography (HPLC) of fenpyr
11、oximate residue in fruit and vegetable. Thi比sstandard i怡sa叩口p剖州li臼bl怡et。thede目tt斩忡rm叫1甘由巾in阳1旧a目tionof f，仿en叩1币pyrox刘ima剖ter陌e吉副iduein c口itrusand cabbage 2 Principle 叫创创圳t盯r-column. The elu挝ewas used for HPt:eUV determintion.，.E皿emak.l;阳dardmethod was used for quantitative analysis. 3 Regents and ma
12、terials 3.2 n-Hexane: Chromatographic f 1:17f i i t二二)I 3.3 Ethyl acetate: Chromatographi!町刮目示飞也 J . _# 3.4 Anhydrous sodium sulfate: Ignite at 650它40r4 h. and 3.5 Sodium chloride solution: 10%. ; water is distilled water. 3.6 Standard of fenpyroximate (CAS. NO. : 111812-58-9): Purity;?99%. 3.7 Stoc
13、k standard solution of fenpyroximate: Suitable amount standard (3.6) was accurately weighed. prepared with acetonitrile at concentration of 100 Ig/mL. Working solution is prepared by properly diluting the stock standard solution to the required concentration with acetonitrile. 3.8 Neutral alumina SP
14、E column: 1 000 mg. 3 mL. 6 SNjT 1977-2007 4 Apparatus and equipment 4.1 High performance liquid chromatography. equipped with UV-Vis detector. 4.2 High-speed blender: 5000 r/min. 4.3 Centrifuge: 4000 r/min. 4.4 Rotary vacuum evaporator. 4.5 5 The edible portions from the represntative samples were
15、taken and b峰ndedin a high speed blender. homogenized thoroughly. ca 500 9 oJ the mixed sample was drawn anclplaced in a clean container as the test sample , sealed and labeled. L-5. 2 Sample storage / I The test sample should be stored atli:rc -4可.In te coursof-.amplingand sample preparation. pre哺ca
16、utions should be taken to avoid 10阳lli，ntionpr any ftors地hichnay cause the change of resi-I ( I , I i due content. 6 Procedure 6. 1 Extraction j 15.00 9 of the test sample was weighed into a 50 mL centrifuge tube. added with 20 mL methanol. and homogenized for 2 min. Sample extract was centrifuged f
17、or 4min at 4 000 r/min and the super natant was filtered. The residue was extracted with additional 20 mL methanol. repeat the centrifu ging and filtration operation. The total filtrate was combined for further clean-up. 6.2 Cleanup The above filtrate was transferred into a 250 mL separatory funnel.
18、 40 mL sodium chloride solution (3.5) and 20 mL n-hexane were added into separatory funne1. The separatory funnel was shaken vig orously for 5 min. let stand to separating. N-hexane layer was transferred into a 100 mL conical flask with stopper. Extra 20 mL n-hexane was added to the aqueous layer to
19、 repeat the operation again. 7 SN/T 1977-2007 The total n-hexane layer was combined into the same conical flask. The flask was added with 2.0 9 anhydrous sodium sulfate. shacked and let stand. N-hexane layer was transferred into a 100 mL con centration flask. The conical flask was washed with little
20、 amount of hexane twice. The total hexane was evaporated to nearly dryness at rotary vacuum eva口。ratorin water bath at 40t. The residue was washed with 3 x 3 mL hexane-ethyl acetate (2 +酌.the above solutions were trans ferred into neutral alumina SPE c咀lumn(2.2.酌.and passed through the column pre-el
21、uted with 5 mL ethyl acetate at a flow rate of 2 mL/min_ The total eluate was collected into a test tube. evaporated to dryness under N, stream in water bath at 4Ot. The residue was re-dissolved in 1 mL mobile phase solution. The solution is filtered through 0.45m micropore filter membrane for LC de
22、termination 6.3 Determination 6. 3. 1 LC condition a) Chromatographic column, ZORBAXSB-C.5m. 250 mm x 4. 6 mm (i. d). or equivalent; b) Mobile phase, acetonitrile-water (85 + 15); c) Flow rate, 0.80 mL/min; d) Detection wavelength, 254 nm; e) Injection volume, 40L 6. 3. 2 LC determination According
23、to the concentration of fenpyroximate in sample solution. the standard working solution with similar peak area was selected. The responses of fenpyroximate in the standard working solu tion and the sample solution should be within the linear range of the instrumental detection. The standard working
24、solution should be randomly injected in between the injections of sample solution of equal volume. Under the above LC operating condition. the retention time of fenpyroximate is a bout 9. 8 min. For chromatogram of the standard see Figure A. 1 in annex A. 6.4 81ank test The operation of the blank te
25、st is the same as the described in the method of determination. but with the omission of sample addition. 7 Calculation and expression of result Calculation the content of fenpyroximate residue in the test sample by LC data processor or accord ing to the formula (1). the blank valne should be subtra
26、cted from the result of calculation E where X=A.c.V As.m X -the residue content of fenpyroximate in the test sample. mg/kg; A -the peak area of fenpyroximate in sample solution; As-the peak area of fenpyroximate in standard working solution; c-the concentration of fenpyroximate in standard work solu
27、tion，g/mL， V-the final volume of the sample solution , mL; m-Mass of the test sample, g. 8 Limit of detection and recovery 8. 1 Limit of detection The limit of detection of this method is 0.02 mg/kg of fenpyroximate 8. 2 Recovery SNjT 1977-2007 ( 1 ) 8.2. 1 According to the experimental data by HPLC
28、-UV , the fortifying concentration of fenpyroxi mate in orange and its corresponding recoveries are , -0.02 mg/kg , the recovery of fenpyroximate is 80.4% -97. 5% , 0.05 mg/kg, the recovery of fenpyroximate is 83.5% -97.1 %, 0.20 mg/kg , the recovery of fenpyroximate is 87.8% -101. 7%. 8. 2. 2 Accor
29、ding to the experimental data by HPLC-UV , the fortifying concentration of fenpyroxi mate in cabbage and its corresponding recoveries are, O. 02 mg/ kg , the recovery of fenp.roximate is 78. 2-96. 0% ; 0.05 mg/闸，the recovery of fenpyroximate is 81.7-97.4% , 0.20 mg/阁，the recovery of fenpyroximate is
30、 87.3-102.5%. 9 SN/T 1977-2007 Annex A ( informative) Chromatog阳mof the standard 3凋-咆, 俑他矿，., J巳 内. .?- li U/ 川?呻F町_,_._ V f!:. Liquid h.(OItog阳nof fenpyroximate d写什牛二.、.;./:! IF 10 中华人民共和国出入境检验检疫行业标准迸出口水果和蔬菜中瞠瞒1I残留置检测方法高效液徊色谱法SN/T 1977-2007 中国标准出版社出版北京复兴门外三里河北街16号邮政编码，100045网址电话，6852394668517548 中国标准出版社秦皇岛印刷厂印刷 开本880X12301/16 印张1字数17千字2007年11月第一版2007年11月第一次印刷印数1-2000 书号，155066.2-18258 定价10.元
