1、EEU 中华人民共和国出入境检验检疫行业标准SN/T 1978-2007 进出口食品中狄氏剂和异狄氏剂残留量检测方法气相色谱白质谱法Determination of dieldrin and endrin residues in food for import and export-GC-MS method 2007-08-06发布2008-03-01实施f新茧t;:飞&1:50 2050 10-20 允许的相对偏差1%土20土25土306.4 空白试验除不加试样外，按上述测定步骤进行。7 结果计算和表述定量离于(m/z)346 380 10 土50用色谱数据处理机或按式(1)计算试样中狄氏剂
2、和1异狄氏剂的残留含量，计算结果需扣除空白值。4 x 生兰二三立一As X m . ( 1 ) 式中zx一一试样中狄氏剂和异狄氏剂含量，微克每千克(g/kg); A一一样液中狄氏剂和异狄氏剂特征单离子的色谱峰丽积3As一一-标准工作溶液中狄氏剂和异狄氏剂特征单离子的色i昔峰zC一一标准工作溶液中狄氏剂和异狄氏剂浓度，微克每升(g/Ll; V 最终样液的定容体积，毫升(mL); m一一最终样液所代表试样量，克(g)。注，计算结果应扣除空白值a计算结果应表示到小数点后两位.8 盟rr定低限、回收率8. 1 测定低限r SN/T 1978-2007 本方法鲤鱼和蜂蜜中狄氏剂的最1定低限为2.5 f1
3、g/kg.)t他样品中狄氏剂的测定低限均为5.0g/kg;异狄氏剂的测定低限均为。/igkg;4 5 SN/T 1978-2007 表3(续)回收率1%样品名称添加浓度1 14000 13000 12000 11000 10000 9000 8000 1000 6000 5000 4000 3000 2000 1000 0 Wi, 60 80 Abundance 5000 4500 4000 3500 370 3000 2500 2000 1500 1000 气um/z 附录B(资料性附录)狄氏剂和异狄氏剂全扫描质i昔图237 346 109 128 狄氏剂NCI会扫描质谱-，-甲_.308_
4、】平380 415 / 12,5 145 .吨445 473 497 521 100 150 250 300 350 400 450 500 图B.2异狄氏剂NCI全扫描质i昔图SN/T 1978-2007 Foreword Annex A of this standard is an informative annexe. This standard was proposed by and is under the charged of the Certification and Accreditation Ad ministration of the people.s Republic o
5、f China E41IHqdat-uab pyi轧FJ3loet峰、电zsugpt , , W Note: This English Version. a translation from the Chinese text , is 501elfor guidance. 9 SN/T 1978-2007 Determination of dieldrin and endrin residues in food for import and export-GC-MS method 1 Scope The standard specifies the determination and conf
6、irmation of dieldrin and endrin residues in foods by Gas chromatography-mass spectrometry. This standard is appli臼bleto the determination and confirmation of dieldrin and endrin residues in rice.mung bean.spinage.string bean.orange.grape. chestnut.vinegar. rose.tea. pork.chicken.pork Ii ver. eel and
7、 honey. 2 Principle The residues in test samples are extracted with acetonitrile-ethyl acetate or n-Hexane-ethyl acetate or ethyl acetate. The extracts are cleaned up by neutral alumina SPE cartridge or florisil SPE cartridge or active臼rbonSPE cartridge. Determination and confirmation made by GC-MS
8、in NCI mode and u sing external standard method. 3 Reagents and materials Unless otherwise specified.all regents used should be analytical grade. water is distilled water. 3. 1 Acetone. 3. 2 Ethyl acetate. 3.3 n.Hexane. 3.4 Acetonitrile: chromatogram grade. 3. 5 Methanol 3.6 Anhydrous sodium sulfate
9、: Ignite at 650C for 4 h. and keep in a tightly closed container. 3.7 Sodium chloride. 3.8 Acetonitrile-ethyl acetate (4+ 1. V / V): Mix 80 mL of acetonitrile with 20 mL of ethyl acetate 3.9 Acetonitrile-ethyl acetate (2 + 3. V / V): Mix 40 mL of acetonitrile with 60 mL of ethyl acetate. 10 SN/T 197
10、8-2007 3.10 n-Hexane-ethyl acetate (1 +4. V!V): Mix 20 mL of n-Hexane with 80 mL of ethyl acetate. 3. 11 n-Hexane-acetone (4 + 1 V! V): Mix 80 mL of n-Hexane with 20 mL of acetone 3.12 Standards and standard solutions: a) Dieldrin: Purity99. O%(CAS: 6057 1) Endrin: Purity二99.0%. (CAS: 72208) , b) St
11、andard stock solution: Accurately weigh adequate amount of dieldrin and endrin standards sepa rately. dissolve with ethyl acetate and prepare a solution of 1.00 mg!mL as standard stock solu tion. Standard stock solution stored at - 18C in a refrigerator, c) Standard middle solution: pipette adequate
12、 amount of standard stock solution.dilute with ethyl acetate to prepare a solution of 10 Ig!mL as standard working solution. Stored at - 18C in a re frigerator; d) Standard working solutions: pipette adequate amount of standard stock solutio门.dilute with ethyl acetate to prepare appropriate concentr
13、ation standard working solutions. Stored at一1日tin a re frigerator 3.13 Neutral alumina SPE cartridge: 2500 mg. 6 mL 3.14 Ftorisil SPE cartridge: 1 000 mg. 3 mL 3.15 Active carbon SPE tubes: 200 mg. 3 mL.ENVI-Carb.or equivalent. 3.16 Strata SDB-L SPE cartridge: Styrene-Divinylbenzene Polymer. 200 mg.
14、 3 mL. or equivalent. 4 Apparatus and equipment 4. 1 Gas chromatograph-mass spectrometry with negative chemical ionization. interface. 4.2 High Speed Homogenizer. 4.3 Ultrasonic water bath. 4.4 Centrifuge: 4000 r!min.6 000 r/min. 4.5 Rotary vacuum evaporator. 4.6 Nitrogen evaporator 4.7 Solid phase
15、extraction vacuum manifold. 11 SNjT 1978-2007 4.8 Vortex mixer. 4.9 Glass tube with scale. 5 mL with stopper. 4.10 Centrifuge tube. polytetrafluoroethylene. 50 mL. 4. 11 Sand funnel. 5 Sample preparation and storage 5. 1 Preparation of test sample I I 5.1.1 spinage.string bean.orang事grape1 Re叫臼町叫叫m叩
16、唰口削l叫叩0问g阳川N怕叫e叩叩p阳paJt1阳m叫叫叫p川内leis m蚓d削叫叫川回M阶阳阶川mt怡a副ine1?川ea叫d.1阳胁a抽b5. 1. 2 rice. mung bean. chestnut., rose. tea la-5.1.5 honey Representative sample. about 500 g. The sample which is not crystallized shall be stirred well to make homogeneous. If the sample is crystallized.it must be warmed in
17、a water-bath below 60C with the sample bottle covered tightly. mix thoroughly when all sample has melted , then cool immediately to room temperature. In the course of samle melting, precautions must be taken to avoid evapora. tion of water from the smple. Place in a clean container.which is labeled
18、and sealed. 5. 2 Storage of sample Tea. honey.viegar.grain and nut should be stord 0C -4C ,vegetable and fruit The test should be 12 SN/T 1978-2007 stord below -18t. In the course of sampleing and sample preparation, precaution should be taken to avolid contamination or any factors that盯laycause cha
19、nge of the residue content. 6 Procedure 6. 1 Extraction 6.1.1 Rice , mung bean Weigh 5.0 9 (accurate to O. 01 g) of test sample into a 50 mL centrifuge tube, add 3 9 of sodium chloride, 20 mL of acetonitrile-ethyl acetate (4 : 1, V / V) (3.8) and 3 9 of anhydrous sodium sul-fate. Homogenize the samp
20、le f命1min, and sonicilte for 10 min.1 After centrifugation for 5 min at 4000 r/min. the supernatant I啡erW8S transfered into a heart-haped flask. Residue was rinsed twice with 10 mL of acetonitrile!.ethyLaCltat豆4盐_:_J.V / V) and cO!1bined the organic solvent. Evap orate the organic solvent to dryness
21、 with r叩a作evaporatqvat45C. Dissolve the residue with /- - -,- j 1.0 mL of n-hexane川eanup./ / / 6.1 , 2 spinage , string bean , orpnSe, grape / Weigh accuratel20.0 9 (accurte to O. 01 g) of test sample into 1mL conical flask with stop-1 min , and sonicate for 10 min. After centrifugation for 5 min at
22、 6 000 r/min. ,transfer 10 mL of the supernatant layer into a 20 mL tube for clean up 6. 1. 4 Eel , chestnut Weigh 5.0 9 (accurate to O. 01 g)of test sample into a 50 mL centrifuge tube. add 10 9 of anhydrous sodium sulfate and 25 mL of acetonitrile-ethyl acetate (4 : 1, V /V). Homogenize the sample
23、 for 1 min , and sonicate for 10 min. After centrifugation for 5 min at 6 000 r/min ,transfer 10 mL of the supernatant layer into a 20 mL tube for a clean up. 13 S:-I/T 1978-2007 6.1.5 Tea , rose Weigh O. 5 9 (accurate to 0.001 g) 01 test sample into a 10 mL centriluge tube , dipped in 1.5 mL water
24、lor 20 min. Add O. 2 9 01 sodium chloride and 0.2 9 01 anhydrous sodium sullate, vortex to mix and add 2 mL x 301 n-hexane.ethyl acetate (4 1, V I V) (3.10) , vortex lor 2 min. centriluge at 4000 r/min lor 3 min. and combined the supernatants lor a c1 ean up. 6. 1. 6 Honey. vinegar Weigh 1.0 g(accur
25、ate to O. 01日)01 test sample into a 10 mL glass tube. add 5 mL water to mix. The diluted solution was passed through a SD8-L SPE column (wash the culumn with 2 mL 01 methanol and 2 mL 01 water belore use) (3.16). and then rinse the column with 15 mL 01 water at a Ilow rate 013 mL/min. dry the column
26、 for 2 min. And then ,elute the column with 2 mL 01 acetone and 3 mL 01 ethyl acetate at a Ilow rate 01 1 mL/min. Collect the eluents into a heart-shaped Ilask. Evaporate nearly to O. 5 mL with rotary evaporator at 45 C lor a clean up. 6. 2 Clean-up 6.2.1 Rice. mung bean , honey, vinegar Setting SPE
27、 vacuum manilold, lill O. 5 9 anhydrous sodium sullate on the top 01 the Ilorisil SPE col umn (3.14). Condition the Ilorisil SPE column )Nith 3 mL 01 n-hexane-acetone(4 1, V / V) (3. 11) belore use. Pass the sample extraction solution through the column and rinse the column with 1.0 mL x 401 n-hexan
28、e -acetone(4 1, V / V) at a Ilow rate 01 1 mL!min. Collect the eluents in a 10 mL glass tube and evaporate under nitrogen to dryness at 45 C. Dissolve the residue and dilute to 1.0 mL with ethyl acetate lor GC-MS analysis. 6.2.2 spinage ,string bean ,orange,grape Setting SPE vacuun manilold. Conditi
29、on the ENVI-Carb SPE column(3. 15) with 2 mL x 201 ethyl ace tate belore use. Pass the sample extraction solution through the column and rinse the column with 2.0 mL x 2 01 ethyl acetate at a Ilow rate 01 1 mL/min. Collect the eluents in a 10 mL glass tube and evaporate under nitrogen to dryness at
30、45 C. Dissolve the residue and dilute to 1.0 mL with ethyl ac etate lor GC-MS analysis. 6.2.3 Pork, chicken , pork liver, Eel , chestnut Setting SPE vacuum manilold. add 1.0 9 01 anhydrous sodium sullate into a neutral alumina SPE col umn (3.13). Condition the Ilorisil column with 4 mL 01 acetonitri
31、le-ethyl acetate (4: 1, V/V) before use. Pass the sample extraction solution through the column and rinse the column with 2. 0 mL 01 acetonitrile-ethyl acetate (4 1, V / V) at a lolw rate 01 1 mL/min. Collect the eluents in a 10 mL glass tube and eva口orateunder nitrogen to dryness at 45C, and recons
32、titute with 1.0 mL 01 n-hex ane-acetone (4 1, V / V) then pass through the Ilorisil column (Iill O. 5 9 anhydrous sodium sullate 14 SN/T 1978-2007 on the top of column bed). Follow the same operation as that described in 6.2.1. 6.2.4 Tea , rose Setting SPE manifold. Condition the ENVI-Carb column wi
33、th 2 mL x 2 of ethyl acetate before use Pass the sample extraction solution through the column and rinse the column with 1. OmL of ethyl ac etate at a flow rate of 1 mL/min. Collect the eluents and evaporate under nitrogen to near O. 5 mL at 45C ,then pass through the florisil column (fill 0.5 9 anh
34、ydrous sodium sulfate on the top of column bed). Rinse the column with 1. 0 mL x 4 of n-hexane-acetone (4 1, V / V) at a flow rate of 1 mLnin. Collect the eluents in a 10 mL glass tube and evaporate under nitrogen to dryness at 45C. Dissolve the residue and dilute to 1.0 mL with ethyl acetate for GC
35、-MS analysis. 6.3 Determination 6.3. 1 GC and MS operating conditions a) Gc column , Capi lIary column , D8-XL日，0.25 mm(i. d. ) x 30 m with 0.25m particle size, or e quivalent, b) Carrier gas, Nitrogen, purity二三99.999% ,Fow rate , 1.5 mL/min,Pressure , 14.6 Psi , c) Injection mode, Splitless,purge o
36、n after 1 min, d) Injection volumn, 1L， e) Injection port temperature, 270C, f) Interface temperature, 280 C, 20 C /min g) Temperature increasing programme, Column temperature,80 C (1.5 min)一一一-一220C(3 min) 5t/min ._o.-. 20t/min 一-一255t:一一一一280t:(4 min) , h) Electron ionization mode, NCI , i) lon so
37、urce temperature, 150C , j) Guadrupole temperature, 150C , k) Solvent protection delay, 5 min, 1) Reaction Gas ,methane, Flow rate ,40% , 15 SN/T 1978-2007 m) Determination mode, SIM, n) Monitor ions(m/z): See table 1. Table l-Monitor ions and Quantitative ions Analyte Time/min Oetected ion(m/z) die
38、ldrn 16.1 237.380.239.346 endrin 16.9 38口.7日.308.3456. 3. 2 GC-MS determination and con Quantitative ions(m/z) 346 380 According to approximate concentratin()f analyte. select the staandard )Norking solution with simi lar response to that of sample solution. The responses pf dieldrin andntJ rin in t
39、he standard working -, ,/ solution and samole solution should be in the line号nffigeof the i阴阳mentaldetection. The standard f - working solution should be injected randomly_)仔betweenthlfnjections of sample solution of equal volume. ./ / According to the operating condition Relative intensity/% Permit
40、ted tolerances/% 6.4 81ank test ,.1 10 :t 50 The operation of the blank test 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 dieldrin and endrin residues in the test sample by GC-
41、MS data processor or according to the formula (1). The blank value should be substracted from the above result of calcula tion: 16 where X=AxcxV As x m X -the residue content of dieldrin and endrin in the test sample.g/kg， A -the peak area of dieldrin and endrin in the sample solution, As一thepeak ar
42、ea of dieldrin and endrin in the standard working solution, SN/T 1978-2007 . ( 1 ) c-the concentration of dieldrin nd endrin in the standard workng!solution.g/L， V-the final volume of the sampl m-the corresponding mass of test sample p 8 Limit of determination and n 8. 1 Limit of determination .h阳.h
43、efi协hefi丁陌时时ef刊f们inals 呵哪polu阳t旧町肌。町r/ F The lmit determnation of this method for dieldrin and何恻刑宁、电eland honeybee is 2. 5 fl9/k9. and 阳limit叫det阳mnat5.0g/kg. 8. 2 Recovery According to the experimental dat 、电飞飞句-地咱.，.，.1Fortified Sample concentrationj (严g/kgl5.0 rice 10 50 5.0 mung bean 10 50 n L-
44、Recovery!% Dieldrin Endrin 80.6-93.2 85.0-99.2 84.5-98.6 86.5-97.4 88.8-99.8 87.9-99.5 82.8-93.6 88.6-98.4 88.5-97.6 88.3-98.2 90.4-98.2 87.6-98.4 17 SN/T 19782007 Table 3 (ntlnued) Fo口ifiedRecovery/% Sample ncentratonl (g!kg) Dieldrin Endrn 5.0 77.2-86.8 80.4-93.4 spmage 10 87.6-95.4 83.2-94.7 50 8
45、9.6-97.1 89.0-98.2 5.0 79.6-91.4 80.8-92.2 string bean 1口88.7-94.4 89.0-94.1 50 91.4-100.2 91.2-97.0 5.0 76.2-87.2 76.8-84.0 orange 10 83.2-92.3 83.3-92.0 50 85.2-95.6 89.4-96.4 5.0 77.4-87.4 77.0-84.4 grape 10 86.5-94.0 83.2-95.7 50 89.8-97.6 88.4-96.2 5.0 80.4-88.4 87.0-100.6 pork 50 80.2-86.9 88.
46、4-102.0 100 85.9-93.2 90.5-94.7 5.0 77.8-91.8 87.2-102.4 chicken 10 83.2-92.3 82.6-92.4 100 85.5-95.2 87.3-96.5 5.0 82.4-89.4 85.8-100.8 阳rkliver 10 82.0-90.5 96.5-103.2 50 86.4-92.7 90.1-96.4 Dieldrin2. 5 89.2-92.4 92.0-97.2 Endrin5.0 eel 10 89.6-94.6 91.2-96.1 50 91.6-96.2 驭。8-97.25.0 83.2-90.8 89
47、.6-106.4 tea 10 80.6-88.0 82.4-96.6 50 88.4-97.0 90.8-95.4 5.0 81.6-92.4 89.6-96.6 chestnut 10 89.5-96.3 90.7-96.3 50 90.4-99.8 90.2-99.2 18 SN/T 1978-2007 Table 3 (continued) Fortified Recove叩/%Sample concentrationl (g!kg) Dieldrin Endrin 5.0 97.4-105.8 84.0-98.0 vrnegar 10 92.1-98.5 95.6-102.2 50
48、92.6-99.0 94.2-99.8 5.0 77.2-91.4 88.2-102.6 rose 10 79.4-93.4 80.4-92.1 50 84.8-97.0 85.4-98.8 2.5 76.8-84.8 8日.8-100.4honey 10 87.5-107.3 87.9-100.6 50 90.4-99.6 91.4-98.8 19 SN/T 1978-2007 Annex A ( informative) GC-MS chromatogram of the dieldrin and endrin standard(TIC) Abundan田5000 EE它-m4500 19
49、.50 20.00 15.5016. 00-.16.50 1旦旦17.5018.00 X ; Y叶(一romatogram(NCJ) of the dieldrin如dendrin standard(TIC) ，.;)_，r j，哗川户19.00 18.50 4000 :; 000 500 0 3500 2500 2000 1500 1000 14.50 14.00 Figure A_ l-GC-MS 20 Abundance 14000 13000 12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 oW!, m/z 60 80 Abundance 5000 -4 500 4000 3500 I 70 3000 2500
