1、中华人民共和国出入境检验检疫行业标准SN/T 1623一2005进出口卫生筷中曝苯咪略、邻苯基苯盼、联苯和抑霉瞌残留量的检验方法液相色谱法Inspection of thiabendazole、o-phenylphenol、diphenyland imazalil in disposable chopsticks for import and export-Liquid chromatography 2005-08-18发布060608000110 2006-02-01实施中华人民共和国发布国家质量监督检验检疫总局中华人民共和国出入境检验检疫行业标准进出口卫生筷中唾苯睐睡、邻苯基苯酣、联苯和抑
2、霉睡残留量的检验方法液相色谱法SN/T 1623-2005 当&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: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2005年11月第一版2005年11月第一次印刷拎书号:155066 2-16467 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:中华人民共和国湖南出入境检验检疫局。本标准主要起草人:戴华、
3、曹建全、李拥军、张华茂、黄志强。本标准系首次发布的出入境检验检疫行业标准。SN/T 1623-2005 I 1 范围进出口卫生筷中曝苯咪瞠、邻苯基苯盼、联苯和抑霉瞌残留量的检验方法液相色谱法SN/T 1623一2005本标准规定了卫生筷中唾苯咪瞠、邻苯基苯酷、联苯、抑霉瞠残留量检验的抽样、制样和液相色谱测定方法。本标准造用于卫生筷中唾苯咪瞠、邻苯基苯醋、联苯、抑霉瞠残留量的检验。2 抽样和制样2. 1 检验批以不超过450万双为一检验批。同一检验批的商品应具有相同的特征，如包装、标记、产地、规格和等级等。2.2 抽样数量抽样数量见表1。表1抽样敢量批量最低取样量骂王900050 9001-36
4、000 125 36001-150000 200 150001-600000 300 二主600001 500 2.3 抽样方法单位为双从货物至少3个不同的部位随机抽取样件，从抽取的样件中逐件开启，按表1规定的抽样数量从样件中抽取样品。将所取全部样品充分混合，装入清洁、干燥的样品袋内密封，并标明标记，及时送实验室。2.4 制样从抽取的样品中再随机抽取样品3双，将附在试样上的碎屑、碎片等清除干净。称取每双筷子的质量ml(准确到0.01g)，再将每双卫生筷劈成长小于1cm、厚约1mm的竹条或削成木片，分别装入清洁密封的试样袋中密封，作为检验样品分别测定。2.5 试样保存将试样于室温保存。抽样和制样
5、的操作过程中，应防止样品受到污染或发生残留物含量的变化。3 测定方法3. 1 方法提要试样中唾苯眯略、邻苯基苯酣、联苯和抑霉瞠用甲醇提取，提取液经过滤后，用配有二极管阵列检测器或紫外检测器的液相色谱仪进行测定，外标法定量。SNjT 1623-2005 3.2 试剂和材料所有试剂除特殊注明外，均为分析纯，水为重蒸锢水。3.2.1 乙睛:液相色谱级。3.2.2 甲醇:液相色谱级。3.2.3 磷酸:85%。3.2.4 十二烧基磺酸铺。3.2.5 唾苯咪瞠标准品:纯度注98.3%。3.2.6 邻苯基苯酣标准品z纯度95.5%。3.2.7 联苯标准品:纯度注99.5%。3.2.8 抑霉瞠标准品z纯度注9
6、9.0%。3.2.9 标准溶液z分别准确称取适量唾苯咪瞠(3.2.5)、邻苯基苯酣(3.2.6)、联苯(3.2.7)或抑霉瞠(3.2.的，用甲醇榕解并定容至100mL，配制成为唾苯咪瞠100g/mL、邻苯基苯酣100t-tg/mL、联苯100吨/mL或抑霉瞠1000g/rnL浓度的标准储备液。根据需要再用甲醇将标准储备液稀释成适当浓度的混合标准工作液。3.3 仪器和设备3.3. 1 高效液相色谱仪配二极管阵列检测器或紫外检测器。3.3.2 t昆匀器。3.3.3 超声波清洗器。3.3.4 离心机:4000r/rnin。3.4 测定步骤3.4. 1 提取、净化准确称取1g(准确至0.001g)试样
7、于30mL具塞离心管中，准确加入5.00rnL甲醇(3.2.2)，具塞混匀，置超声波清洗器中超声30rnin，然后在旋涡混匀器上棍匀3rnin，于2000 r/rnin离心5min，上清液过0.45m微孔捷、膜后，上HPLC测定，外标法定量。3.4.2 测定3.4.2. 1 色谱条件a) 色谱柱:ODS柱(25crnX4. 6 mrn ID，5m)或与之相当的柱。b) 流动相:称取十二烧基磺酸铀0.681g，加入350rnL甲醇、50rnL乙睛、100rnL水和1mL磷酸溶解，过0.45m微孔滤膜。c) 流速:0.5rnL/rnin。d) 检测波长:唾苯咪瞠、邻苯基苯酣、联苯的检测波长为247
8、nr川抑霉哇的检测波长为226nrn。e) 进样量:10Lof) 柱温:400C。3.4.2.2 色谱测定根据样液中唾苯咪瞠、邻苯基苯酣、联苯和抑霉瞠的含量情况，选定峰面积相近的标准工作、溶液。标准工作溶液和样液中唾苯眯嗖、邻苯基苯盼、联苯和抑霉哩的响应值均应在仪器的检测线性范围内。对标准工作溶液和样液等体积参插进样测定。在上述色谱条件下，唾苯咪瞠、邻苯基苯盼、抑霉哇、联苯的保留时间约为8.1 rnin、8.6min、11.1 min和14.0rnin。标准品色谱图参见附录A中图A.l和图A.203.4.3 空白实验除不加试样外，按上述测定步骤进行。3.5 结果计算和表述用色谱数据处理机或按式
9、(1)、式(2)计算试样中唾苯咪瞠、邻苯基苯酣、联苯或抑霉瞠的含量，计算2 SN/T 1623一2005结果需扣除空白值。Ax Cs X v j = A s X m2 X 1 000 x. mj . ( 1 ) 。=AXCsxV As X m2 ( 2 ) 式中z1一-每双试样中唾苯咪瞠、邻苯基苯酣、联苯或抑霉睡的残留量，单位为毫克每双(mg/双); 2一-试样中唾苯咪瞠、邻苯基苯酣、联苯或抑霉瞠的残留量，单位为毫克每千克(mg/kg); A一一样液中唾苯咪瞠、邻苯基苯酣、联苯或抑霉睡的峰面积，单位为平方毫米(mm2); Cs-一标准工作液中唾苯咪瞠、邻苯基苯酣、联苯或抑霉睡的浓度，单位为微克
10、每毫升(g/mL); A. -标准工作液中唾苯咪瞠、邻苯基苯酣、联苯或抑霉瞠的峰面积，单位为平方毫米(mm2); V一一样液最终定容体积，单位为毫升(mL);mj一一每双筷子的质量，单位为克(g);m2一一最终样液所代表的试样质量，单位为克(g)。4 测定低限、回收率4. 1 测定低限本方法唾苯咪瞠、邻苯基苯酣和联苯的测定低限为1.0 mg/kg;抑霉略的测定低限为5.0mg/kgo 4.2 回收率4.2. 1 卫生筷中唾苯咪哇添加浓度在1.0 mg/kg、5.0mg/kg、10.0mg/kg、25.0mg/kg和50.0mg/kg 时回收率范围在81%92%。4.2.2 卫生筷中邻苯基苯酣添
11、加浓度在1.0 mg/kg、5.0mg/kg、10.0 mg/kg、25.0 mg/kg和50.0 mg/kg时回收率范围在81%92%。4.2.3 卫生筷中联苯添加浓度在1.0 mg/kg、5.0mg/kg、10.0mg/kg、25.0mg/kg和50.0mg/kg时回收率范围在82%92.1%。4.2.4 卫生筷中抑霉哩添加浓度在5.0mg/kg、10.0mg/kg、25.0mg/kg、50.0mg/kg和125.0mg/同时回收率范围在81%105%。3 SN/T 1623一2005附录(资料性附录)标准晶色谱圄A go-TH桐沓目由.由E桶叫mMR且非亭。.寄送锵剧Chl247nm m
12、Abs 150 10 5 。20 唾苯咪睡、邻苯基苯酣、联苯混合标准晶的液相色谱固15 10 5 。圄A.14 SN/T 1623-2005 Chl226 nm mAbs 150 v一才80.=副EMW豪100 。丁50 110 20 15 10 5 5 抑霉睡标准晶的液相色谱固圈A.2SN/T 1623一2005Foreword Annex A of this standard is an informative annex. This standard was proposed by and is under the charge of The Certification and Accr
13、editation Admin istration of the Peopl白Republicof China. This standard was drafted bHunan Entry-Exit Inspection and Quarantine Bureau. The standard was mainly drafted by Dai Hua. Cao Jianquan. Li Yongjun. Zhang Huamao and Huang Zhiqiang. This standard is a professional standard for entry-exit inspec
14、tion and quarantine promulgated for the first time. Note, This English version ,a translation from the Chnese text , is solely for guidance. 6 SN/T 1623-2005 Inspection of thiabendazole、o-phenylphenol、diphenyland imazalil in disposable chopsticks for import and export 一Liquidchromatography 1 Scope T
15、his standard specifies the methods of sampling , sample preparation and determination of thiaben dazole、o-phenylphenol、diphenyland imazalil residues in disposable chopsticks by liquid chromatogra phy. This standard is applicable to the determination of thiabendazole、o-phenylphenol、diphenyland imaza
16、liI residues in disposable chopsticks. 2 Sampling and sample preparation 2. 1 Inspection lot Each inspection lot should not be exceed 4 500 000 pairs. The characteristics of the臼rgowithin the same inspection lot,such as packing ,mark,origin ,specifi cation and grade etc. should be the same. 2. 2 Qua
17、ntity of sample taken (See table 1) Table 1 Quantity of sample taken unit: palrs Number of packages in an inspection lot Minimum number of packages to be taken 三二900050 9001-36000 125 36001-150000 200 150001-600000 300 二主600001500 2. 3 Sampling procedure Draw at least three park of packages at rando
18、m , open the packages one bone， and take the samples according to table 1. The combine primary sample is fully mixed, placed in a clean sample canister , sealed , labeled and sent to laboratory in time. 2. 4 Preparation of test sample Take three pairs of chopsticks of the mixed primary sample at ran
19、dom , make the chopsticks surface smooth ,Weigh every pairs of chopsticksCaccurate to 0.01 g). Then split it long ca. 1 cm , ca. thin 1 mm. Each portion is placed in a clean container as the test sample , which is then sealed and 7 SN/T 1623-2005 labeled. 2. 5 Storage of test sample The test samples
20、 should be stored at room temperature. In the course of sampling and sample prepa ration.pre臼utionsmust be taken to avoid contamination or anfactors which macause the change of residue content. 3 Method of determination 3. 1 Principle Thiabendazole、o-phenylphenol、diphenyland imazalil residues are ex
21、tracted from the disposable cops ticks by methanol .and the extract is fpassed through membrane filter. The solution is used for LC determination by PAD or UV. External standard method is used for quantitative measurement. 3. 2 Reagents and materials Unless otherwise specified.all the reagent used s
22、hould be analytical grade. water is distilled water. 3.2. 1 Acetonitrile: HPLC grade. 3. 2. 2 Methanol: HPLC grade. 3.2.3 Phosphoric acid:85% 3. 2. 4 Sodium laurysulfonate. 3.2. 5 Standard of thiabendazole: purit注98.3%.3. 2. 6 Standard of o-phenylphenol: purit注95.5%.3.2.7 Standard of diphenyl: purit
23、y ;?:99. 5%. 3.2.8 Standard of imazalil: purit注99.0%.3.2.9 Stock standard solution: Accurately weigh an adequate amount of thiabendazole (3. 2. 5) o-phenylphenol(3.2.邸.diphenyl(3.2. 7) or imazalil(3. 2. 8) standard.dissolve with methanol and prepare a solution of 100g/mL as standard stock solution o
24、f thiabendazole, o-phenylphenol or di phen叭，and1 000g/mL as standard stock solution of imazali l. According to the requirement ,dilute the standard stock solution with methanol as the mix standards working solution. 3.3 Apparatus and equipment 3.3.1 HPLC equipped with PAD (photodiode array detector)
25、 or UV detector. 8 SN/T 1623-2005 3. 3. 2 Mixer. 3.3.3 Ultrasonic bath. 3.3.4 Centrifuge: 4000 r/min. 3. 4 Procedure 3. 4. 1 Extraction Weigh accurately 1 9 of the test sample (accurate to O. 001 g) into 30 mL centrifuge tube with stop per. Add 5.00 mL methanol (3.2.2) and sonicate for 30 minutes in
26、 an ultrasonic bath (shaking at least every 10 min). Let cool the sample to room temperature, centrifugalize at 2 000 r/min for 5 min,and filter through 0.45m membrane filter. The solution is ready for HPLC determination, using external standard method. 3. 4. 2 Determination 3. 4. 2. 1 HPLC operatin
27、g condition a) Column: ODS (25 cm x 4.6 mm i. d. ，5m) or equivalent. b) Mobile phase: Dissolve O. 681 9 sodium laurysulfonate in 350 mL methanol , 50 mL acetonitrile , 100 mL water and 1 mL phosphoric acid , then fi Iter through O. 45m membrane filter. c) Flow rate:O. 5 mL/min. d) Detector wavelengt
28、h: thiabendazole、o-phenylphenoland diphenyl at 247 nm; imazalil at 226 nm. e) Injection volume: 10L. f) Column temperature:40oC. 3. 4. 2. 2 HPLC determination According to the approximate concentration of thiabendazole , o-phenylphenol ,diphenyl or imazalil in the sample solution ,select the standar
29、d working solution with similar peak area to that of the sample solution. The responses of thiabendazole, o-phenylphenol , diphenyl or imazalil in the mix standard working solution and sample solution should be within the linear range of the instrumental detection. The mix standard working solution
30、should be randomly injected in-between the injections of the sam ple solution of equal volume. Under the above operating condition , the retention time of thiaben dazole is about 8.1 min ,o-phenylphenol is about 8.6 min , imazalil is about 11.1 min and diphenyl is about 14.0 min. For chromatogram of
31、 the standard ,see Fig. A.1 and Fig. A. 2 in annex A. 3.4. 3 Blank test The operation of the blank test is the same as that described in the method of determination but without addition of sample. 3. 5 Calculation and expression of the result 了hecalculation of the residue content of thiabendazole ,
32、o-phenylphenol , diphenyl or imazalil in the test sample is carried out by HPLC data processor or according to the formula( 1) or the formula(2) , 9 SN/T 1623-2005 the blank value should be subtracted from the result of calculation: ，-A x c. x V o _ _ _ x m, A. x m2 x 1 000 ., Ax c. x V 2 = A. x m2
33、、B/aE, ，、. . . . . . . . . . . . . . . . . . . . . . . . ( 2 ) Where, 1一一theresidue content of thiabendazole,o-phenylphenol ,diphenyl or imazalil in the test sam ple per pairs (mg/pairs); 2一一-theresidue content of thiabendazole.o-phenylphenol.diphenyl or imazalil in the test sam ple Cmg/kg) ; A-the
34、peak area of thiabendazole ,o-phenylphenol.diphenyl or imazalil in the sample solution Cmm2) ; c.一-theconcentration of thiabendazole. o-phenylphenol , diphenI or imazalil in the standard working solution (g/mL) ; A.一thepeak area of thiabendazole.o-phenylphenol.diphenyl or imazalil in the standard wo
35、rk ing solution (mm2); V -the final volume of the sample solution (mL); m,-the mass of the test sample per pairs (g); m2-一一thecorresponding mass of the sample in the final sample solution (g). 4 Limit of determination and recovery 4. 1 Limit of determination The limit of determination of thiabendazo
36、le，o-phenIphenol and diphenI is 1.0 mg/kg,and imazalil is 5.0 mg/kg. 4. 2 Recovery 4.2. 1 According to the experiment data. the fortifying concentrations 1. 0 mg/kg、5.0 mg/kg、10.0 mg/kg, 25. 0 mg/kg and 50.0 mg/kg of thiabendazole in dsposable chopsticks. and their corre sponding recoveries ranges a
37、re 81 % .92%. 4.2.2 According to the experiment data. the fortifying concentrations 1. 0 mg/kg、5.0mg/I甸、10.0 mg/kg、25.0mg/kg and 50.0 mg/kg of o-phenylphenol in disposable chopsticks.and their corre sponding recoveries ranges are 81 % .92%. 4.2.3 According to the experiment data. the fortifing conce
38、ntrations 1. 0 mg/kg、5.0mg/kg、10.0 mg/kg、25.0mg/kg and 50. 0 mg/kg of diphenyl in disposable chopstcks .and their correspond ing recoveries ranges are 81 % .91 %. 4.2.4 According to the experiment data. the fortifying concentrations 5.0 mg/kg、10.0mg/坤、25.0 mg/kg , 50.0 mg/kg and 125.0 mg/kg of imaza
39、lil in disposable chopsticks,and their correspond ing recoveries ranges are 81%.105%. 10 SN/T 1623-2005 Annex A ( Informative) Chromatogram of standards 咱的04时Hbgoz且弓时的由国-。E乱-AEZ且占喃喃0.由-。目ERSChl 247 nm mAbs 150 10 5 。20 mm Chromatogram of thiabendazole,o-phenylphenol and di由enylstandard 15 10 5 。Figure A.1 mCON-伺NFFHZ08.=-M苟且EChl226 nrn mAbs 150 SN/T 1623一2005100 50 飞口lJn20 15 10 侵极必究书号:155066.2-16467 定价:10.00元* 版权专有Chromatogram of imazalil standard Figure A. 2
