【商检行业标准(SN)】 进出口动物源性食品中氟甲喹残留量检测方法 液相色谱－质谱/质谱法

中华人民共和国出入境检验检疫行业标准SN/T 1921—2007
进出口动物源性食品中氟甲喹残留量检测方法
液相色谱-质谱/质谱法
Determination of rcsidue of flumcyuinc in foodstuffs of animal origin forimport and export-I.C-MS/MS methoc2007-05-23发布
中华人民共和国
国家质量监督检验检发总局
2007-12-01实施
本标准录A机断录为资料性附求
本标准由区家认证认可监督管理委员会提出并力口，本标滩山中州检验恰鞍科学研究院负支起草。不标滩主要起草人：影涛，安姆、国伟、个晓娟、严不，综历、唐章、储晓刚。本标准系首次发布的出入境检验检疫行业标准。SN/T1921—2007
1范围
进出口动物源性食品中氟甲喹残留量检测方法液相色谱-质谱/质谱法本标准规定广动物源性食品中氟中峰残量液相色谱质谱/质谱测定方汰本标准适压弱物组织、内脏.亞、鱼利虾中冠丰噬残留量的检测2试样制备与保存
2.1动物组织、内脏,鱼和虾
SN/T 1921—2007
从原始样品取出有代丧性样品约590需，用纽织捣碎机充分捣碎混勾，均分成两份，分别装人洁净容露件为试样，密村，准标明标记。将试样胃」一18冷冻避北保行2.2蛋
从原始样品圾出有代表性样品药500g，去壳厂用凯织擒控机搅拌充分混勾，沟分成两份，分别装人洁净容器作为试样.密毒，标实标记，将试样置」4冷越避光保存2.3奶
从原始样品取出右代表性样品约500惠，用纠织揭谛机充分准匀.均分成两分，分别装人洁净容器准为流样.密封，并标明求记：将试样置于1℃冷藏逃光保存在制样的换作过程守成防而样品污染或发牛残南物含量的变化3方法提要
用磷酸盐缓冲液和乙精提取试样中戏留的氯旦嗪，经正已烷与除脂肪、固州率驳杆烂化后采用液相色谱质谱/质谱检测，外标法定员。4试剂和材料
除非见有说明所有流剂为分析绝水为大离子水残相站纯度的求，4.1乙睛：高效液色谱级、
4.2正亡烷：高效浚相色谱级：
4.3币萨：高效液相色级。
4.4磷酸.
4.5磷酸氢钾
4.6氢氧化钠。
氮水;25头
币酸：高效液相色谐级。
乙酸铵
4.1CZ腈饱和的正烷:间10℃m Z腈中加人109 =nL正烷,充分振荡后,静置分层,取1.层乙哨备用：
4.11磷酸盐缓冲液(c.051mol/L，pII7.40.2)：准确移取250mLC.2mol/L磷酸氧许溶液利1:7.5 n-I.0,2mol/I.氢氧花钠水溶液丁：I.溶员瓶中，可加水 50c ml斥磷骏或氢氧花销凋节pH7.2—(.2,作水定容至T.。
SN/T 1921—2007
4.12SPE洗悦液：尚 7ml.甲醇中训人25 ml.氨水混勾备用。4.13c.「杀半废水落波（含0.tmt:in./.乙酸铵）:准确取|ml.H酸和称改.0386只乙铵：「「萃量瓶用水荠「「
4.14标准品：氧甲噬纯厚99炎
4.15标维佑备液：确称取适量氛币唾标准品（精硫至0.00℃1 ）,用2清溶解，制成浓度为100 m/1的标准诺备举渡，18冷冻避光保存3个月。4.1E中间标准溶液：通移取1ml.标准储备液于10 ml.容量中出乙晴定容至刻度,配制成度为10！11/L的中间标准溶液，4℃冷藏避光保存1个H。4.17标准工作溶液：根据需要斥C.为巾校水溶液（=.13)稀释皮适而浓度的标准工作溶被，现而现配。
C固相幸取.3ml..00mg，或相当名，4. 19
微孔滤膜：0.2热m，有机椎，
氛气:继浅密用.99%
5仪器和设备
5.1液相色谱质谱/质谱仪：配备电喷雾离子源(LS1)5.2红织碎机
5.3均质器：10 C00:/in
5.4摄荡器。
离心机:10 co9 -/nizr.
氮吹仪：
5.7涡动仪，
5.8趋,i波水浴：
容呆瓶：「「.,[C ml.
5.1C潮料离心管：50l.
分液漏斗：12511L
5.12品筒:50 rml.。
刻变流管：10ml.
6测定步骤
6.1提取
6. 1.1动物组织、内脏、鱼和虾称取药2 g试样(精铺至3. 01 g)于50 ml.塑料离心管中,加人 18 ml.磷骏盐缓冲液(1.:1)和2 ml.乙情,用均质露以[c0or/min为质2srin片，再以[の0c0min离心10 min=收集上清液于12Fml分液漏习-中，加1人20) :ml.乙情饱和后的正已烷（-.[C）,振荡IC mit:,静胃 2) rmim,收集全部下层液体于5.简中.用水定容3Cml.
6.1.2蛋和奶
称取约2 或样(精劲9. 01)于50rml 塑料离心管,加人 18 nml. 磷酸盐缓冲液(4._1)和2rl乙清，用振器振部提域20 mir后，再以13 c03 r/min离心2℃ min，余下梁作同6.1.1。6.2净化
Ci:固相萃收注（2. 18)依次用 3mL中醇、6 11:L水预游洗眉，准确转人15 ImL栏昂提取液(6.1)，月3 rmT. 水进行淋洗,弃么;再用 3 mT. SPF 洗脱液(2. 12)进行洗脱,收集洗脱液于 1C -mT.刻度试管中 ,整个固相举取净化过程挤制流述不超过2=nL/=nirn，跳脱液在10℃小用氮气吹干：残留物用1 rnl2
SN/T1921—2007
0.【%币酸水溶液(，13)落解，涡旋【mir后,过0.2 u:微孔滤膜(.19）,供仪器测定：6.3测定
6. 3. 1 液相色谱条件
6. 3. 1. 1 谱柱:C,J:0) t:it:×2. . mm(内径), p:,或粘当若6.3.1.2流动和:7.肺C.1%干酸水辨液（5~55.体积比）6. 3. 1. 3 杜温:30℃,
6. 3. 1. 4流速:C. 2 ml./ min.6.3.1.5逊样量：10 l.
6. 3.2质谱条件
参沈谢求A
6. 3. 3 液相色谱 质谱/质谱测定根据样液中氟+哗的含量，选定浓度相近的标准！作溶液，待测样液中氟中喉的恂成佳应在仪器检测的线性范用内对标准1作溶液及样液(5.2)等体积参折过样测定：在上述仪器条件下，氧中哮保菌时间约为1.1min.液相色谱-质谱/质谱负满图参见附录B图B.6.3. 4阳性样品的确证
按照「述条件测定样品利标准工作液，如呆检测的质量色谱峰保案对间与标准工作液-致，允许编差小于2.5%;定性离对的树对十度与浓变杆当标准工作液的树对十度致,杆对度编差不超过表1的观定，则可判断样品中才在相应的被测物表1定性测定时相对离子丰度的最大充许偏差相刘离下度《)
允许的相遍差（)
6. 4 空自试验
除不川试样外，按上述测定步骤迹行。结果计筑和表达
试栏中的袭留含量,按式(1)计算：20450
X-A.XmX1 00C
式中：
试样京氟中峰綫率含量，单位为寇克年千克(ri/k);样液守氟中怪的峰而积；
际雅一作溶液方氟中嗑的峰而积：[C20
标准：作溶液京氟中峰浓变，单位为纳克每毫升(ng/rl）样液最终定容体积，单位为宅升(ml.)；样液所代表最终试样的质量，单位为克（&）江：计笋结果需游审门直扣除。8测定低限和回收率
8.1测定低限
本标雅的测定低限为:0.c09 5 rmg/k8.2回收率
鸡肉：添加浓度0.0005mg/kg：可改率为%0.6%~106.0：添加浓度0,005 r/k-回收率为 76.0%~-1c5,6%50
SN/T1921—2007
添加浓度0.0mg/k:周收率为89.1%100.9%码径：添川浓度0.00 ：mg/kg，可改率为72.c%~[0%.0%，添加浓度0.605 rm/kg.向收率为82.0%~1c4.6%：添加浓度0.05mg/kg.回收率为81.6~-95.2薪叶：添加浓度0.c09 5rmg/kg，回改率为76.0%~-106.0%：添加浓度0.c05mg/k，叫收率为82.0%~98.5%；添川浓度0.05 mg/kg,[收率为81. 0%-~95. 8%牛奶：添加浓度0.6005mg/kg.可改率为76.0%~104.0次：添加浓度0.c05rmg/kg.回收率为74.0%96.5%；添加浓度0,05 mg/kg:收率为 86,8%-.98.4鱼：添加浓度0.c00Emg/kg，叫改率为78.c%100.0%：添加浓度0.c0 mg/kg，收率为82.0~[c4.c%：添加浓度0. 05 tm/kg.同收率为84. 2%~102. 4%添加浓度0,609 51mg/k，可改率为76,6%~105.0；添加浓度0.005rng/kg.回收率为72.0关~-96.0关；添加浓度0.05mg/k：周收率为81.8%95.2%化密
氣同座
商子用丁于是母
二离方式
纠管中压
源泥发
去溶剂温度
锥孔气流
大辖剂气流
硅撞气水
监测模式
附录A\i
（资料性附录）
质谱条件
质谱条件
多反应监测条件
子离子
.20℃
微气.100./h
微气.600 /h
SN/T1921—2007
氢气,2. 2010 Pa
密设应监
维孔电点,V
能流能母/
1）附录A所列参数是在Wn:cr Qur:ru Prmirr序讲仪1完戏的,比处列出试验用仪器型受为了垃供参学,并不数及商业月的敲厨标准使用样尝试采川不同厂家改型号的仪辈：3www.bzxz.net
SV/T1921—2007
附录Ｈ
（资斜性附录）
标准物质色谱图
262-244
262202
氟甲喹液相色谱-质谱/质谱多反应监测色谱图Ji.ne
Foreword
Annax A and Anncx B of this standard arc informativc annexcsSN/T1921—2007
This standard was proposed by and is under the jurisdiction of the Certification and AccreditationAdministration of the People's Republic of ChinaThis slandard is drafled by Chinese Acaderny of Inspectiar and Quaranline.Main drafters of this standard are: Peng Tao, An Juan, Guo Wei,Li Xiaojuan, Yan Mao, Xu Li, TangYingzhang and ChuXiaagang
This standard is a professional standard for Entry-Exit inspection and quarantine of the People's Re-public of China promulgated for the first time.Nate: This English Version+ a translation from the Chinese text, is solely for guicdenceSV/T1921—2007
Determination of residue of flumequinein foodstuffs of animal origin forimportandexport-Lc-MS/MSmethod1 Scope
This standarcd specifies the determinatian of flumequine residue in animal origin foodstuffs for importand export by Lc-Ms/MS.
This standard is applicable to the determination or Tlumequine residue in animal tissue: offall: egg:milk, fish, and shrimp.
2 Sample preparation and storage2.1 Anirnal tissue, offall, fish and shrirnpAll primary samplc is rcduccd to 5a0 g as the represcntative samplc, which is blcnded and homagc-nized, and then divided into two equal portions. Each portion is placed in clean containers as the testsarmple, which is sealed and labeled. The test sample shauld be stored below in -18'C avoiding sun-light.
All primary samplc is rcduccd to 500 g as thc rcprcsentativc samplc, which is shclled and homagc-nized, and then divided into two equal portions. Each portion is placed in clean containers as the testsarnple, which is sealed anid labeled. The test sarnple should be stored below in 4'℃ avoidirig sur.light.
All primary samplc is rcduccd to 50d g as thc rcprcscntativc sample. which is homogenizcd, andthen divided into two equal portions. Each portion is placed in clean containers as the test sample.which is sealed and labeled. The test sarnple should be stored below in 4'c avoiding surlight.In the course of sarnple preparation, precautions should be taken to avoid contamiriatiori or ariy fac-torswhichmaycauscthochangeofrcsiduccantcnt.S
3Principle
SN/T 1921—2007
The flumecuine residue in the test sample is extracted with phosphate buffer and acetonitrile. Aftercleaned up with n-hexane and C- SPE cartridge, the residue is then delermined by LC-Ms/MS. andquantificd bycxternalstandardmethod.4 Reagents and materials
Unless otherwise specilied, all regents used are A. R. : and pure \water\ is redistilled water.4.1
Acetonitrile:HPLC grade
N-hexane: HPLC grade.
Methanol:HPLC grade.
Phosphoric acid.
Potassium dihydrogen phosphate.Sodium hydroxide.
Ammonia: 25%,
Formic acid: HPLC grade.
Armmonium acetale.
N-hexane saturated with acetonitrile: add 100 mL n-hexane into 100 mL acetonitrile. mix ad-equately, then wait for delamination. use the substrate layer.4.11Phosphatebuffer(0.05mol/LpH7.4±0.2):add250 mL0.2 mol/Lpotassium dihydrogenphosphate solution, 197.5 mL0.2mol/L sodium hydroxide solution and500 mL water into1L Volu-rrnelric flask. then adjusl pH7. 4= 0. 2 with phasphoric acid or sodiurni hydroxide solulion, diluled lc1 L with water.
SPE eluent solution: add 25 mL 25% ammonia into 75 mL methanol, mix adequately.4. 130. 1% formic acid solution (cantaining 0. 5 mmol/L ammonium acetate): acdd 1 mL formicacid and 0. 038 6 g ammonium acetate into 1 L volumetric flask, then dilute to 1 L with water.4.14Flumcquinc standard:Purity99%9
S.V/T 1921—2007
4. 15 Standard stock solution: accurately weigh an adequate amount of flumequine standard, dis.solve in acetonitrile and prepare a salutian of 1oo mg/L as the standard stock solution, stored below-18C avoiding sunlight forthreemonths.4. 16 Middlc standard solution: accurately transfer 1 mL standard stock solution into 10 mL volu-metric flask, then make up to graduation with acetonitrile. prepare a solution of 10 mg/L as the mid-dle standard solution. stored below 4'c avoiding sunlight for a month.4.17
Standard working solution: according to the reguirement. dilute middle standard solution tcappropriate concentration with 0. 1% formic acid solution (4, 13)just before use.4.18
Cis SPE carlridge: 3 mL, 500 mg, or equivalenlMernbrane filter: 0. 2 μm, organic type.Ntrogen: purity.99. 999%.
Apparatus and equipment
Liquid chromatography-tandem mass spectrometry, ecuipped with electrospray ion source.Tissue blender.
Homogenizer: 10 0a0 r/min.
Centrifugc: 10 0ap r/min.
Nitrogen evaporator.
Vortex mixer.
Ultrasonic water bath.
Volumetric flask: 1 L, 10 mL.Plastic centrifuge [ube: 50 mL.Separating funnel: 125 mL.
Volumetric cylindcr: 50 mL.
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