SN T 1770-2006 进出口粮谷中抑虫肼残留量测定方法.液相色谱法.pdf

1、中华人民共和国出入境检验检疫行业标准SN/T 1770-2006 进出口粮谷中抑虫阱残留量测定方法液相色谱法Determination of tebufenozide residues in cereals for import and export-Liquid chromatographic method 2006-04-25发布060905000037 2006-11-15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粮谷中抑虫脚残留量测定方法液相色谱法SN/T 1770-2006 奇峰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:10

2、0045网址电话:6852394668517548 中国标准出版社秦皇岛印刷厂印刷9峰开本880X 1230 1/16 印张l字数22千字2006年8月第一版2006年8月第一次印刷印数1-2000提书号:155066 2-17034 定价10.00元前言本标准的附录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黑龙江出入境检验检疫局负责起草。本标准主要起草人:杨长志、康庆贺、田丰、韩广源、高勇。本标准系首次发布的出入境检验检疫行业标准。SN/T 1770-2006 SN/T 1770-2006 1 范围进出口粮谷中抑虫胁残留量测定方法液相色谱法本标准规定

3、了进出口粮谷中抑虫脚残留量检验的抽样、制样和液相色谱测定方法。本标准适用于进出口大米中抑虫脚残留量的检验。2 抽样和制样2. 1 检验批以不超过200t为一检验批。同一检验批的商品具有相同的特征，如包装、标记、产地、规格和等级等。2.2 抽样数量按式(1)计算抽样袋数:a =.fN 式中:a 抽样袋数;N一一-全批袋数。注:a值整数，小数部分向前进位为整数。2.3 抽样工具. ( 1 ) 2.3. 1 金属单管取样器:全长55cm(包括手柄)，直径1.5 cm2. 0 cm，沟槽长度应超过袋对角线长度的一半。2.3.2 取样铲或取样勺。2.3.3 分样板。2.3.4 盛样器:筒或袋，可密封。2

4、.3.5 分样布或适用铺垫物。2.4 抽样方法2.4. 1 倒包抽样从堆垛的各部位随机抽取2.2规定的应抽样件数的10%(每批一般不少于3袋)，将袋口缝线全部拆开，平置于分样布或其他洁净的铺垫物上，双手紧握袋底两角，提起约成45。倾角，倒拖1m以上，使袋内货物全部倒出。检查货物的外观、气昧、有无发霉、变质等，并查袋内和袋间品质是否均匀。确认情况正常后，用取样铲随机在各部位抽取样品，立即将样品倒入盛样器内。每袋抽取样品数量应基本一致。2.4.2 袋内抽样按2.2规定的应抽样件数(扣除倒包抽样袋数)，在堆垛四周上、中、下各层以曲线形走向随机抽取。将取样器(2.3.1)管槽朝下，从每袋一角依斜对角方

5、向插入袋内，然后将管槽旋转朝上，抽出取样器，立即将样品倒入盛样容器内。每袋抽取样品数量应与2.4.1基本一致。每批样品总量应不少于4kg。2.4.3 大样缩分合并倒包和袋内抽样所取全部样品，倒于分样布上，用分样板按四分法缩分样品至不少于2峙，倒SN/T 1770-2006 入盛样器内，加封后标明标记，并及时送交实验室。2.5 试样制备将样品按四分法缩分至1kg，全部磨碎并通过20目筛，y昆匀，均分成两份作为试样。分别装入洁净的容器内，密封，标明标记。在抽样和制样过程中，应防止样品受到污染或发生残留物含量的变化。2.6 试样保存将试样于OOC40C保存。3 测定方法3. 1 方法提要试样中残留的

6、抑虫缩、定容后，用配有紫外检3.2 试剂与材料除另有规定外，所用试剂均为分析纯，水为3.2. 1 甲醇:色谱纯。3.2.2 乙腊:色谱纯。3.2. 3 丙酬。3.2.4 二氯甲炕:色谱纯。3.2.5 正己:皖:色谱纯。3.2. 6 元水硫酸铀:经63.2.7 中性氧化铝:层析3.2.8 硅胶:层析用，60昌伊100目.foC烘2h. 3.2.9 抑虫脚标准品:纯度二三98%。性氧化铝和硅胶柱净化，浓3.2.10 抑虫阱标准洛液:准确称取适量的抑虫脚标准品，用乙睛配成浓度为100g/mL的标准储备液。根据需要用流动相稀释成适当浓度的标准工作液3. 3 仪器与设备3.3.1 液相色谱仪:配有紫3.

7、3.2 粉碎机。3. 3. 3 振荡器。3. 3. 4 旋转蒸发器:配3.3.5 氮吹仪。3. 3. 6 具塞锥形瓶:2503. 3. 7 分液漏斗:2503. 3. 8 滤膜:0.45m。3. 3. 9 层析柱:25 cm X 6 g中性氧化铝、4g无水硫3.3. 10 元水硫酸纳柱:8.03. 4 测定步骤3.4. 1 提取g无水硫酸铀、1.0g硅胶、正己炕(1十9)预淋洗。硫酸铀。称取试样约20g(精确到O.1 g)于250mL具塞锥形瓶中，加入100mL丙酣+水(80+20)，在振荡器上振荡提取30mi口，过滤于250mL浓缩瓶中。用30mL丙酣洗涤锥形瓶于同一浓缩瓶中，在450C水浴

8、中用旋转蒸发器浓缩至约20mL。将浓缩液转移到250mL分液漏斗中，再用60mL水洗涤，合并于同一分液漏斗中，加入2X50mL二氯甲炕充分振摇萃取2min，静置分层。将二氯甲皖层通过无水硫酸铀柱(3.3.10)脱水于250mL浓缩瓶中，在450C水浴中用旋转蒸发器浓缩至近干，再用氮气流吹2 干。用4mL丙酣+正己烧0+9)溶解残渣。3. 4. 2 净化SN/T 1770-2006 将上述溶液移入层析柱(3.3.9)中，用20mL丙酣+正己皖0+9)洗涤浓缩瓶，待液面降至无水硫酸铀表面时，加入柱内淋洗，弃去流出液。用20mL丙酣十正己烧0+1)洗脱，收集全部洗脱液于100 mL浓缩瓶中，在450

9、C水浴中用旋转蒸发器浓缩至近干，再用氮气流吹干。准确加入1.0 mL流动相(3.4.3.1)溶解残渣，经O.45m滤膜过滤后供液相色谱测定。3.4.3 测定3.4. 3. 1 液相色谱条a) 色谱柱:Nb) 流动相:乙睛c) 流速:1. 0 mL/ d) 柱温:400C;e) 检测波长:234nm; f) 进样量:20L。3.4. 3. 2 液相色谱测定根据样液中抑虫阱残留量情应值均应在仪器检测线性下，抑虫阱的保留时间约3. 4.4 空白试验除不加试样外，均按3.5 结果计算和表述用色谱数据处理机或按式(2)计算试样中抑虫脚残留量，计算结果应将空白值扣除。式中:h s 标准工作搭液一标准工作溶

10、液V 样液最终定m 最终样液所4 测定低限、回收率4.1 测定低限本方法的测定低限为4. 2 回收率样品中抑虫脚的添加浓度在0.500mg/kg时，回收X一主羊三羊vh s X 在0.050mg/kg时，回收率为87.0%; 在0.025mg/kg时，回收率为85.6%。. ( 2 ) 3 SN/T 1770-2006 附录A (资料性附录)标准晶色谱圄-、mAU-11.231 15 童倒霉基1 10 5 。5 。2 4 6 自10 12 14 16 18 t/min 圈A.1抑虫Jl#标准晶渣相色i普圄4 SNjT 1770-2006 Foreword Annex A of this sta

11、ndard is an informative one. This standard was proposed by and is under the charge of National Regulatory Commission for Certi fication and Accreditation Administration of the Peoples Republic of China. 丁hisstandard was drafted by Heilongjiang Entry-Exit Inspection and Ouarantine Bureau of the Peopl

12、es Republic of China. The main drafters of this standard are Yang changzhi ,Kang qinghe,tian feng and Han guangyuan ,Gao yong. This standard is an Inspection and Ouarantine professional standard promulgated for the first time.1) 1) Note: This English Version.a translation from the Chinese text. is s

13、olely for guidance. 5 SN/T 1770-2006 Determination of tebufenozide residues in cereals for import and export-Liquid chromatographic method 1 Scope This standard specifies t zide residues by liquid 2 Sampling and sample 2. 1 Inspection lot Each inspection lot determination of tebufeno-es in rice for

14、import and export. The characteristics of the cargo within the same inspection lot , such as packing , mark , origin , spec ification , grade ect. , should be the same. 2. 2 Quantity of sample ta The number of bags to where a一一-numberof bags N一一一totalnumber of Note: If value a is with 2. 3 Sampling

15、tools 2.3.1 Metallic sampler: length: longer than half the d 2.3.2 Sampling shovel or ladle. 2.3.3 Plate for quartering. 2.3.4 Sample container: Can or bag , which can be sealed. 6 ula ( 1): ( 1 ) the integral part of a er: 1.5 cm 2.0 cm; groove SN/T 1770-2006 2.3. 5 Cloth ( or other suitable materi

16、al) sheet: For sample dividing (quartering). 2.4 Sampling procedure 2.4. 1 Sampling by emptying out Draw 10 percent of the at random. Unseam and tight two corners of t 2.4. 2 Sampling from inside Draw the samples from in 2.4. 1) as follows: middle and lower parts bags) at any part of the pile or oth

17、er clean sheet). Grasp rd for ca. 1 m until all uniform within and between up the sample with a shovel the number of bags drawn from the bags of the upper, sampler ( 2. 3. 1) , with its groove facing downwarc-甲吨擎酶n倒出t叩叶跚且nit恤且自由蜡1800，draw out the sampler and promptlpour the sample into a container.

18、The quantity of sample drawn from each bags should be basically the same as in 2. 4. 1. The total weight of the sample of ea lot should Pour all the samples ( plate bquartering. PI 2. 5 Preparation of Grind with a grinder to portions as the samples. and sample preparation , cause change of the resid

19、 2. 6 Storage of test sample The test sample should be stored OoC _40C . to not less than 2 kg with a e laboratory in time. Iy and divide into two equal 1. In the course of sampling on or anfactors that may 7 SN/T 1770-2006 3 Method of determination 3. 1 Principle The tebufenozide residues in the te

20、st sample are extracted with acetone-water. The extract is ex tracted with dichloromethane, and clean up with an alumina and a silic gel column. The elutes solution is evaporated. The residues is dissolved with acetonitrile-water and determined by HPLC With UV detector, using external standard metho

21、d. 3. 2 Reagents and material Unless otherwise specified , all regents are analytically pure. Water is redistilled water. 3. 2. 1 Methanol: LC grade. 3.2.2 Acetonitrile: LC grade. 3. 2. 3 Acetone. 3.2.4 Dichloromethane: LC grade. 3.2.5 N-hexane: LC grade. 3.2. 6 Anhydrous sodium sulfate: Ignite at 6

22、50 oC for 4 h and store in a desiccator. 3. 2. 7 Neutral alumina: For chromatograph， 100 mesh. 200 mesh , Ignite at 300 oC for 4 h and store in a desiccator. 3.2.8 Silica gel: For chromatography, 60 mesh-100 mesh , Heat at 1100C for 2 h and store in a desiccator. 3.2.9 Tebufenozide standard: Purity9

23、8%. 3.2. 10 Tebufenozide standard solution: Accurately weigh an adequate amount of tebufenozide standard , dissolve in acetonitrile and prepare a solution of 1009 /mL as the standard stock solu tion. According to the requirement , dilute a standard working solution of appropriate concentration with

24、mobile phase. 3.3 Apparatus and equipment 3.3.1 High performance liquid chromatograph equipped with UV-detector. 8 SN/T 1770-2006 3.3.2 Grinder. 3. 3. 3 Shaker. 3.3.4 Rotary evaporator with 100 mL ,250mL evaporated flask. 3.3. 5 Nitrogen evaporator. 3.3.6 Conical flask: 250 mL, with stopper. 3. 3. 7

25、 Separator funnel: 250 mL. 3.3.8 Membrane filter: 0.45m. 3.3.9 Chromatographic column: 25 cm x 2. 0 cm (i. d. ) Glass column , pack with a little degreased cotton at the bottom of column and add in sequence 2 9 of anhydrous sodium sulfate, 1 9 of silica gel , 6 9 of neutral alumina , 4 9 of anhydrou

26、s sodium sulfate. Pre-rinse the column with 20 mL of ace tone-n-hexane (1 + 9) before use. 3.3. 10 Column of anhdrous sodium sulfate: 8.0 cm x 1. 5 cm( i. d) ,packed with 5 cm height of an hydrous sodium sulfate. 3. 4 Procedure 3.4. 1 Extraction Weigh 20 9 (accurate to O. 1 g) of the test sample int

27、o a 250 mL conical flask , add 100 mL of ace tone-water (80 + 20) , and shake for 30 min with a shaker. Filter the extract into a 250 mL evaporated flask. Wash conical flask into the same a 250 mL evaporated flask with acetone. Evaporate the extract ca. 20 mL with rotary evaporator in a water-bath b

28、elow 45 oC. Transfer the extract and wash evapo rated flask with 60 mL water into a 250 mL separator funnel. To the separator funnel , add 2 x 50 mL of dichloromethane, then shake violently for 2 min and let stand to separate clearly. Pass the dichlo romethane layer through an anhydrous sodium sulfa

29、te column (3.3.10) to remove the water, let drain into a 250 mL evaporated flask , Evaporate the dehydrate dichloromethane to near dryness with rotary evaporator in a water-bath below 45 oC ,then blow to dryness under a nitrogen flow. Dissolve the residue by adding 4 mL of acetone-n-hexane (1 +9). 3

30、. 4. 2 Clean up Rinse the above solution into the chromatographic column (3.3.9). Wash the evaporated flask with 20 mL of acetone-n-hexane (1 +9)into the column ,when the liquid levellowers to the upper surface of anhydrous sodium sulfate, then discard the effluent. Elute with 20 mL of acetone- n-he

31、xane (1 + 1) and collect all the eluate in a 100 mL evaporated flask. Evaporate the eluate to near dryness with ro-9 SNjT 1770-2006 tary evaporator in a water-bath below 45 oC , then blow to dryness under a nitrogen flow. the residue is dissolved accurately with 1 mL mobile phase(3. 4. 3. 1) , filte

32、red through a 0.45m membrane and ready for LC determination. 3. 4. 3 Determination 3.4.3.1 a) LC column: N b) Mobile phase : a equivalent ; c) d) Column temperature: 40 oC ; e) Detector wavelength: 234 nm; f) Injector volume: 20L 3. 4. 3. 2 LC determination According to the estimated the standard wo

33、rking soluti tebufenozide in the standard sample solution , select ution. The responses of be in the linear range of the instrumental detection. II咱二剖and佛非*omng啥。1日悦。句附情丁币口edrandomlin-between the injections of the sample solution of equal volume. Under the above chromatographic condition , the reten

34、tion time of tebufenozide is ca. 11 . 2 min , For the chromatogram of tebufenozide standard , see annex A. 3. 4. 4 81ank test The operation of the blank without addition of sample. 3. 4. 5 Calculation and The calculate the content of formula (2) , the blank value where: -霄，哇-一-X一一-theresidue content

35、 of tebufenozide the test samples , mg/ kg ; h-the peak height of tebufenozide in the test sample solution , mm; hs一一一thepeak height of tebufenozide in the standard working solution , mm ; of determination , but r or according to the ( 2 ) C一一一theconcentration of tebufenozide in the standard working

36、 solution，g/mL; V一一-thefi nal volume of sample solut ion , mL ; m一一一thecorresponding mass of test sample in the final sample solution , g. 10 SN/T 1770-2006 4 Limit of determination and recoverv 4. 1 Limit of determination The limit of determination of this method is O. 025 mg/kg. 4. 2 Recovery Acco

37、rding to the experimental data , the fortifying concentration of tebufenozide in the sample and its corresponding recoveries are: 一一一0.500mg/阔，the recovery is 94.8% ; 一一0.050mg/kg, the recovery is 87.0%; 一一一0.025mg/炮，therecovery is 85. 6%. OON|ohhFHZm SN/T 1770-2006 Annex A ( informative) Chromatogram of the standard mAU 11.231 可胃口口-言LF20 15 10 5 。18 16 14 12 10 自6 4 2 。phu n m 侵权必究书号:155066 2-17034 定价:10.00元 版权专有liquid chromatogram of the tebufenozide standard Figure A.1 SN/T 1770-2006