1、sm 中华人民共和国出入境检验检疫行业标准SN/T 1989-2007 进出口食品中丁M井残留量检测方法气相色谱-质谱法Determination of daminozide residues in food for import and export Gas chromatography-mass spectrometry method 2007-08-06发布2008-03自01实施能mFUPL叫吨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辽宁出入境检验检疫局负责起草。本标准主要起草人:董振
2、霖、田苗、杨春光、徐凤敏、赵景红、董伟峰、于灵、王宏伟。本标准系首次发布的出人珑检验检疫行业标准。SN/T 1989-2007 1 范围进出口食品中丁酷酣残留量检测方法气相色i昔-质谱法本标准规定了食品中丁航脐残留量的气相鱼谱-质谐检测方法。SN/T 1989一2007本标准适用于花生、大米、大豆、欧芹、苹果、鱼、鸡肉、茶叶、蜂蜜中丁酷脐残留最的测定。2 方法提要试样巾的丁酬残留物用水址主主旦旦，水杨应衍生化J硅胶固相萃取柱净化.用气相包谱质谱仪进行测定，外标法定量。./飞J / 3 试剂与材料除另有规定外，所用试剂均为分阔，水兑?宽阔Ji3. 1 正己烧2色i苦纯。3.2 氢氧化的。3.3
3、氯化的。3.4 冰乙段。3.5 水杨脏。3.6 4 仪器与设备4. 1 气相色谱仪，配质量选择性检测器。4.2 水蒸气蒸饱装置。4.3 离心机:5 000 r/min. 4.4 旋转蒸发仪。4.5 振荡器。4.6 超声波仪。4. 7 因相萃取柱2硅胶.6mL. 1000 mg.或相当者。4.8 具塞三角烧瓶:250mL。4.9 具寒试管:25mL。一一SN/T 1989-2007 5 试样制备与保存5. 1 水果和蔬菜类取苹果、欧芹等有代表性样品约500g，将其可食用部分切碎后，用捣碎机将样品加工成浆状。混匀，装入洁净的容器内，密闭并标明标记.一18C以下保存。5.2 粮谷和坚果类取大米、大豆
4、、花生等有代表性样品约500g，用粉碎机粉碎并通过孔径20日圆孔筛。1昆匀.装入洁净的容器内，密闭并标明标记。4C以下保存。5.3 动物源食品取鸡肉、鱼等有代表性样品约500g用绞肉机绞碎，混匀.装人洁净的容器内，密闭并标明标记。一18C以下保存。5.4 蜂蜜和茶叶取蜂蜜和茶叶有代表性样品约500g，装人洁净的容器内.密闭并标明标记。4C以下保存。6 测定步骤6. 1 提取粮谷、坚呆:称取试样25g(精确至0.1g)于250mL具塞三角烧瓶中，加入100mL 水，盖;莓，在振荡器上振荡30mino在4000 r/min离心10min，将上iff液倒入蒸馆瓶中，加入氢氧化纳溶液(3.9) 50
5、mL.用少量水冲洗瓶壁，连接水蒸气茶馆装置，缓慢加热至沸腾。用事先加入乙酸溶液(3.8)3mL、水杨l150L的25mL具塞试管接收馆出液，收集馆出液约15mL。蔬菜、水果、水产品、肌肉组织、茶叶、蜂蜜=称取试样25g(精确到O.1 g)于250mL具寒三角烧瓶中，加入100mL水，盖霉，在振荡上振荡30min。将提取i在直接倒入蒸馆1瓶中，加入氢轼化饷溶液(3.9)50 mL，用少量水冲洗瓶壁，连接水蒸气蒸馆装置，缓慢加热至沸腾。用事先加入乙酸溶液(3.8) 3 mL，7j(杨百50L的25mL具塞试管接收t贸出液，收集惚出:在约15mL。6.2 衍生接收试管50C超声30min冷却至室温后
6、，加入约3g氧化纳.5mL正己烧，振摇1min。静宜分层，上层正己烧相供净化或GC-MS部定(蔬菜、水果、茶叶、蜂蜜样品按6.3步骤净化，其他样品直接进样分析)。取适当体积的1，1-二甲基联氨标准溶液按上述步骤衍生化，供GC-MS测定。6.3 净化对于蔬菜、水果、茶叶、蜂蜜梓1日，用硅胶团相萃取柱(SPE)净化。固相萃取柱用10mL正己烧预淋洗，准确移取正己烧溶液(6.2)4mL至固相萃取柱上，用10mL正己炕淋洗.再用5mL乙酸乙隙正己烧溶液(3.7)淋洗，10mL乙酸乙li5正己统溶液(3.7)洗脱。收集洗脱液，50C水浴减压浓缩至下，用正己炕定容至4mL.供GC-MS测定。6.4 测定6
7、.4. 1 气徊色i晋-质i昔条件a) 色i肯柱，DB-5MS石英毛细管柱，30mXO. 25 mm(内径)，眼厚O.25m，或相当者;15C I b) 鱼i苦柱温度，70C一一二二.270C(5 min) , c) 进样口温度，240C，d) 色-质i昔按口温度，280C，e) 载气=氮气，纯度二99.995%,1. 0 mL/min , f) 进样量，1L，2 SN/T 1989-2007 g) 进样方式:无分流进样，0.75min后开阀gh) 电离方式:EI，i) 电离能量:70eV , j) 测定方式z选择离子监测方式(SIM), k) 监测离子(m/z):定量164.定性149、16
8、3、165，l) 济剂延迟:6min 6.4.2 气相色谱-质i曹测定6.4.2. 1 定量测定根据样液中丁眈脐含量情况.选定浓度相近的1.1-二甲基联氨标准工作溶液衍生化。1，1二甲基联氨标准工作溶液衍生物和样液中1 1二甲基联氮衍生物响应值均应在仪器检测线性范围内。对1.1二甲基联氨标准工作溶液衍生物和样液等体积参插进样进行定。在上述色消条件下.)，卜二甲基联氨衍生物保留时间约为8.17min，1，1二甲基联氨标准品衍生 物SIM色谱图参见附录A中图A.l，6.4.2.2 气相色i昔质i曹确证标准溶液衍生物及样液均按6.4.1规定的条件进行测定，如果样液巾与标准溶液衍生物相同的保留时间有峰
9、出现，则对其进行质谱确证。经确证分析被视J物SIM色i古峰保留时间与标准样品相一致，并且在扣除背景后的样品谱蜀中，所选择的离子均出现g选择离子(m/z)164、149、163、165(其丰度比100 : 8 : 7 14)与标准样品衍生物相关离子的相对丰度致，相似度在允许偏差之内(见表)，1.1 工甲基联氨标准品衍生物SIM质i苦图参见附录A中图A.2。表1GC-MS确证相对离子丰度的最大允许偏差相对丰度(基峙)/%EI.GC-MS(相对)/%50 土1020-50 土1510-20 ic20 10 士506.5 空白试验除不加试样外，均按上述步骤进行。7 结果计算和表述用包谱数据处理机或按式
10、。)计算试样中丁股脐的含量，计算结果需将空白值扣除。X=x c X V X 2. 67 As Xm ) l ( . . . . . . . . . . . . . . . . . . . . . . . . . . . . 式中zx一一试样巾T酌胁的含量._I(t位为毫克每千克(mg/kg), C一一标准工作液中)，1-二甲基联氨衍生物的浓度，单位为微克/毫升(lg/mLl, A 样品溶液中1，1二甲基联氨衍生物的峰而积;As一一标准工作溶液中1，1二甲基联氨衍生物的峰面积;V 样液.lil终定容体积.单位为毫升(mLl, m 占1终样?夜所代表的试样置，单位为克(g)，2.67-1，1-二甲
11、基联氨与丁it脐换算系数。B 测定低限和回收率8. 1 测定低限本方法的测定低限为0.01mg/kg, 3 SN/T 1989-2007 8.2 回收率样品的添加浓度及回收率数据见表2.表2样品的添加浓度及回收率数据添加浓度/(mg/kg)样品名称0.01 0.02 0.05 花生90.5-100.3 89.2-95.6 88.6-92.4 大来91. 2-100. 7 86.5-95.6 86.2-92.2 大旦90.7-97.自91. 9-99. 8 87.8-94.4 欧芹85.7-96.2 81. 3-92. 6 82.7 -90.2 苹果. . ;-6-37 85.0-93.2 鱼8
12、. 7-94. 9 S7.S;:纯.90.2-凹.0鸡肉FFO916l SZ. Z.，7.p 93.1-98.2 茶叶96.0-104.7 /吁97;:1,r 03. 5 98.9-106.8 蜂蜜.3-103. .)!;-自.4-89.285. 492. 7 / / J/djfr b/f/庐/ 4 SNjT 1989-2007 附录(资料性附录)标准品色谱图A abundance 14 000 12 000 ，/m;n jli/ /旧/ 矿/ i j y 飞J/10 000 ahundance 100 60 40 80 8 000 4 000 6 000 2 000 图A.l20 m/z 1
13、74 163 149 。16B 162 156 150 144 5 1, 1-二甲基联氨标准晶衍生物选择离子监测模式(SIM)质谱图图A.2SN/T 1989-2007 Foreword Annex A of this standard is an informative annex. This standard was proposed by and is under the charge of Certificati口nand Accreditation Administra tion of the People s Republic of China. This standard was
14、 drafted by Liaoning Entry-Exit Inspection and Quarantine Bureau of the People s Republic of China. The main drafters of this standard are Dong Zhenlin , Tianmiao. Yang Chunguang. Xu Fengmin. Zhao jinghong. Dong Weife吨.Yu Ling. and Wang Hongwei. This standard is an Entry-Exit inspection and quaranti
15、ne professional standard promulgated for the first time. Note: This English version ,a translation from the Chinese text. s solely for guidance. 6 SN/T 1989一2007Determination of daminozide residues in food for import and export-Gas chromatography-mass spectrometry method 1 Scope This standard specif
16、ies the determination of daminozide residue by gas chromatography-mass spec trometry in foods for import and export. This standard is applicable to the determination of daminolzide residue in foods for import and ex port. 2 Principle The test sample is extracted with water. The extract is distilled
17、with steam , and then derivatized with salicylaldehyde. After cleaned up by silica gel SPE , the analyte is determined by gas chromatog raphy equipped with mass spectrometry detector, using external standard method. 3 Reagents and materials Unless otherwise s口ecified，all the reagents used should be
18、analytical grade, water is distilled wa-ter. 3. 1 Hexane: HPLC grade. 3.2 Sodium hydroxide. 3.3 Sodium chloride. 3.4 Acetic acid. 3. 5 Sal icylaldehyde. 3.6 Ethyl acetate: HPLC grade 3.7 Elute solution:ethyl acetate + hexane (3+97 , V/V) . 7 SN/T 1989-2007 3.8 Acetic acid solution(10%. V/V):Transfer
19、 10 mL acetic acid into a 100 mL beaker and add ap proximately 90 mL water, Mix well. 3.9 Sodium hydroxide solution (50%. m/ V): Weigh 500 9 and dissolve in about 1 L water. cool and mix well 3.10 1.1-dimethyl hydrazine standard (C，比陀.CAS: 57-1 4-7) :purity98%. 3.11 Stock standard solution of 1. 1-d
20、imethyl hydrazine: Accurately weigh 0.1 9 (accurate to 0, 1 mg) of 1 .1-dimethyl hydrazine and dissolve in 2 mL a呗eticacid solution (3.8). Transfer to a 1 L volumetric flask with water to pllpar!Whe，_:;tock.，sta口出昭，:;01以i阻碍。f100g/mL in concentration. Dilute the standard stock solutio with water to t
21、he required concntration as the standard working solution. The stock solution shold be stored in dark below 4t an自canbe used within six months 4 Apparatus and equipment ,1/# / /P / JY 4. 1 Gas chromatography: equippE!6 with massslective detector (MSDl. 4.2 Steam-distillation apparatu.-4.3 Centrifuge
22、: capable of 5 000 r/min. 4. 4 Rotary evaporator. 4.5 Shaker. 4.6 Ultrasonic. 4. 7 i闯d句峭吵叫g归阮户川e酌盯叫:si剖i4.8 Conical flask: 250 mL. with stp间旦-旷4. 9 Glass test tube: 25 mL. with stopper. 5 Sample preparation and storage 5. 1 Fruit and vegetable 1户-.-._-，-吨J / J Reduce the fruit or vegetable sample of
23、 apple. parsley etc to ca 500 9 by quartering. blend and mix thoroughly in a high speed blender. place in ciean containers. seal and label. Store them in - 18t refrigerator. 8 SN/T 1989-2007 5.2 Cereal and nut Reduce the cereal or peanut sample of rice. soybean. peanut etc to ca 500 9 by quartering.
24、 grind thoroughly and let pass through a 20 mesh sieve. mix thoroughly and place in clean containers. seal and label. Store them in 4 c below 5.3 Animal original food Reduce the animal original food sample of chiken. aquatic product to ca 500 9 by quartering. blended and homogenized thoroughly i甲占1电
25、Ihs阳创hl剧由阳制啊啊时闸门containers.seal and label. Store them in -18t refrigerator. Procedure 6. 1 Extraction shaker. The extract is poured into round bottom flask. add 50 mL of sodium hydroxide solution (3.如.connectthe steam-distillation apparatus. Heat the water in the round bottom flask to boil. collect
26、the distillate about 25 mL in a Glass test tube with 3 mL acetic acid solution (3.8) and 50L salicylaldehyde. 6.2 Derivatization After the distillation. place the collection tube in the ultrasonic water bath. ultrasonic the distillate for 30 min at 50C. Cool to ambient temperature. add ap口rox.3日NaCI
27、. 5 mL hexane. shake for 1 min 9 SN/T 1989-2007 The hexane layer is prepared for clean up or GC-MS determination. 6. 3 Clean-up Vegetab怡，fruit , tea , honey:Rinse the silica gel column with 10 mL hexane before use , pipet 4 mL of the hexane layer solution (6.2) into a silica gel column. Wash with 10
28、 mL of hexane , follow with 5 mL of hexane solution (3.7). Then elute with 10 mL of hexane solution (3.7) , collect the elute Concentrate the elute to dryness at 50t , dissolve the residue and adjust the volume to 4 mL with hexane for GC.MS determination 6. 4 Determination 6.4. 1 GC-MS operating con
29、dition a) Chromatographic column: 30 m x O. 25 mm (i. d. ), O. 25m film thickness , DB.5MS silica capil lary column , or equivalent; 15t Imin b) Column temperature: 70t一一一-270t(5 minl; c) Injection temperature: 240t; dl Interface temperature: 280t; el Carrier gas: Helium , purity;三99.995%，1.0mL/min;
30、 f) Injection volume: 1L; gl Injection mode: Splitless , purge on after 0.75 min; hl Electron ionization mode: EI; j) lonization energy: 70 eV; jl Determination mode: SIM mode; k) Selected monitoring ion ( m/z ) : Determined by 164, confirmed by 149, 163 , 165 , i) Solvent delay: 6 min. 10 SN/T 1989
31、-2007 6.4.2 GC-MS determination 6.4.2. 1 Quantitative determination According to the approximate concentrate of daminozide in the sample solution , select the standard working solution of 1 , l-dimethyl hydrazine with similar peak area to that of sample solution. The re sponses of 1, l-dimethyl hydr
32、azine derivative in the standard working solution and sample solution should be within the linear range of the instrumental detection. The standard working solution of 1 , 1-dimethyl hydrazine derivative should be randomly injected in-between the injections of the sam ple solution of equal volume Fo
33、r GC-MS SIM chromatogram (TIC) of the 1, l-dimethyl hydrazine derivative rention time is 8.17 min ,standard see figure A. 1 in annex A. 6.4.2.2 GC-MS qualitative determination According to the operating condition assigned in 6.4. 1 , analyze the standard solution and sample so lution. If there is a
34、peak appeared at the same retention time for both of the sample solution and standard working solution , the GC-MS confirmation test should be conducted. If the retent旧ntimes of sample chromatogram peaks are consistent with the standard. and after subtracted background noise, the relative intensity
35、ratios of each qualitative ions are also consistent with similar concentra tion standard , selected monitoring ions (m/z) 164, 149, 163.165 (relative abundance ratio, 100 8 : 7: 14)and the similarity degree of their relative abundance ratio in permitted tolerance(See table 1) We can confirm that the
36、re are corresponding analyte in the sample. For GC-MS spectrum of the 1 ,1-dimethyl hydrazine derivative standard , see figure A. 2 in annex A. Table l-Maximum permitted tolerance for relative ion intensities using a range of mass spectrometric techniques Relative intensity(base peak)/% EI-GC-MS( re
37、lative) /% 50 :t 10 20-50 士1510-20 土2010 士506. 5 81ank test The operat旧nof the blank test is the same as that described in the method of determination but with omission of the sample addition. 7 Calculation and expression of result Calculation the content of daminozide in the test sample by data pro
38、cessor or according to formula 11 SN/T 1989-2007 (1) the blank value should be subtracted from the above result of calculation A x c x V x 2. 67 X=AV一.( 1 ) where X-content of daminozide in the test sample. mg/kg, c-the concentration of 1 l-dimethyl hydrazine in the standard working solution.g/mL; A
39、 -the peak area of 1. l-dimethyl hydrazine derivative in the sample s而lution，As-the peak area of 1.1-dimethyl hydrazine derivative in the standard working solution, V-the final volume of the sample solution. mL , m-the corresponding mass olte lest sample -tnefriiil samp(e solution. g , 2. 67-the fac
40、tor for daminOzideloW1-二di旧1刊me4牛-一二一一一甲町啤且i / -,/ Jf rvfp 8. 1 Limit of determination /扩/The Iimit of determination of this 11 m内j8. 2 Recovery Table 2-Fortifying levels CJiLisa油iJKLh 内l咱忖!(时/Sample name 0.01 的20.05 peanut .:;:-斗f89 -95.6 I 88.6-92.4 rlce 91 86 !5-95. p 86.2-92.2 50bean 90.7-97.8 9
41、1.9 ;:;99.8 87.8-94.4 parsley 85.7-96.2 81.3-92.6 82.7-90.2 apple 84.6-95.1 85.6-95.7 85.0-93.2 fi5h 86.7-94.9 87.8-96.7 90.2-98.0 chicken 88.0-94.6 92.3-97.8 93.1-98.2 tea 96.0-104.7 97.1-103.5 98.9-106.8 honey 94.3-103.7 80.4-89.2 85.4-92.7 12 SN/T 1989-2007 Annex A ( informative) GC-MS chromatogr
42、am CTIC) of the 1 , 1-dimethyl hydrazine derivative standard abundnnce 14 000 12000 10 000 8000 /一/fry-hydmzine dcrivatve l ,t.dimeth 6 000 4 000 2000 怕m 川1 d 阳n附川e刊d164 / IJJJi -83可爱442jt6 Figure A. 1-GC-MS chro abundance 100 60 40 20 80 m.z 114 16 149 。168 162 156 150 144 13 Figure A. 2-SIM mass spectrum of 1, 1-dimethyl hydrazine derivative standard 中华人民共和国出入境检验检疫行业标准迸出口食品中丁酷脐残留量检测方法气相色i晋-质谱法S-I /T 1989-2007 9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，1045阿址电话6852391668517518 中国标准出版社秦皇岛印刷厂印刷, 开本880X1230 1/1 6 印张1.25 字数26千字2007年II月第一版2007年II月第一次印刷印数1-2000唔书号155066.2-18274定价12.元
