【商检行业标准(SN)】 出口乳及乳制品中氢化可的松残留量检验方法

中华人民共和国进出口商品检验行业标准SN 0700-1997
出口及乳制品中氢化可的松
残留量检验方法
Method for theleternination of hvdrocortisoneresidues in milk aad milk products for cxport1997-12-01发布
中华人民共和国国家进出口商品检验局1998-04-01实施
SN070G-1997
本标准是损据CB/T1.1—19934标准化工作导第1单元：标准的起草与表述规购英1部分；标注端写的基本规定+及SN/T0001—15S5出口商品中药、善药残量及生物毒素检验方法标范编写的基本规定\的要求进行编写的。其中淡定方法是象考国内外有关女献，经研究，改逆和验证后而制定。本标准司时制定了推样和制样方法。测定低限是根据巨际上对牛乳中氛化可的松留查的最高限些和测觉与法的灵敏度而制定的，本标雅的附录A为提示的附录。
本标准由中毕人民未和国国家进出商品检险局提出并归口。本标准动中华人民共和国浙江出口商品检验局负卖起革，本起准主要帜草人断学彬张需，洗小生，本标准系首次发布的行业模准：1范围
中华人民共和国进出口商品检验行业标准出口乳及乳制品中氨化可的松
残留量检验方法
Mcthod fur thc delermination oe hydrocortlsonereaidnts in milk and milk produels far exportSN0700
本标准规定了出口乳及就制品中氢化可的松残南量检验的抽样，齿样和激相负谱测定克法。牢标准适时乎出口归牛乳中氢化可的松弹量的检验。2抽祥和削样
2.1、检验据
以不超过50000率火一抢验
同一检验我的商品应具有相后的特链，如包装、括记，产地，梵格和等级等，2.2拍样数量
批感，瓶
10000下
10 00~20(00
20 001-~30 000
30 00:-~20 000
40 001-50 000
2.3样方法
低抽伴数瓶
按2.2规定的抽样瓶数随机描取原始品，标明标记，及对送交实验室2.4试样制务
将所政蛇伴品充分混与，分出约20ml.，再均分成两份作为试样。分别装入法净的奔器内，谢封，标明标记。
2.5试样保长
将试详于··5C以下避光保存，性，在抑样改制样的举作过理中，必筑药止样品受到污染或发生氧化与的松合量的变化，3测定方法
3.1方法提要
用二氧干烷提取式样中残留的氧化可药松，有机整氢氧化铁熔激性落净化，报缩至下，残渣溶于流动相，所得熔液用正己烷除前后，用配有紫外检测举的限相色诺议测定，外标法定息。3. 2 减剂和利料
除另有规定外，所用试齐均为分新统，水为游缩水。中华人民共和国固家进出口商品检验离1997-12-01批满1998-04-01实施
3.2.1审尊：液相色谱较。
SN0700—1997
3.2.2二氯甲烷，单燕衡，收巢3941它懒分。3.2.3正己烧，重蒸焰。
3.2.4氢氧化钠榨液:0.1mal/L水筛液。3.2.5硅土55。
3.2.6氢化可的格标准品，纯度2S9.0%。3.2.7驾化可的松标准铬激：准通称敢遗量的粒化定的相标推录，用流动相配成陈度为100格/m.的标港备液，羽根据需要用流或相裙释成适当淤度的标催工作裕液，3.3仅器和设备
3.3.1高效换扫免谱仪，配有外检测器。3.3.2旋转蒸发器。
3.3.3高心机，
3.3.4解端混匀器。
3. 3. 5 空气究浓缩装置,
3.3.6硅菌上柱：260mmX2mm（内轻），具活赛玻璃柱，挂底块少量脱指提，上装56健土Ca. 2. 53.
3. 3.7分液斗:250 mL,
3.3.6离心管具塞，5ml.
3.3.5微量注谢器：25L。
3.4测步媒
3.4.1提取和你化
称取试样约1Cg（料至0.1g）于烧杯中，加入约10g硅净十.摄挣沟可，效些10mir将混均物袭人硅满上柱中（5.3.6）入4m二氯甲统，前15mi后，月分液斗滴mL二瓶中烷十注中控制商入和流出速度为2--1ml./min。收金部流出该了寻一分滤漏平中，依次用30mL氢氧化钠籍较、30mL水洗漆，分白有机相。将有机于35C水浴中用能转蒸发解游统至约2-ml.特涨缩液划入5工耳磨离心管内，用2×1mL二氟干烷球拖壁，流疫并入海心单内，以空气流液缩率十。弹碗加入1ml.流效用(挡确至0.02ml.,左航混泥动登上滤分23nin用3×1:mt.芷已烷油提或次，每次划1tmie.以300c/min高心3min，并用尖喷减管穿去正已烷。余液供液色诺测定。3.4.2测定
3.4.2.1液书色潜条件
a）色错住，(os-柱，135mir.>:4.mn（内登），粒径5rmsb）流动明：年醇-水（4455)
）流速，2.0mL/n
）测定波长：245h
e）柱：室盟，
进样量：2℃l..
3.4.2.2游担色净制
根据样液中氢化可劳格含些情沉巡定雄面识相近的标准工作销摘，标准工作落控和样拨实化可的松响应的应在仪能检测载生范国，对标准工作培滤和伴液等体积等插逆栏测定：在上运色谱条牛下，氢传可的松的保南川约为76点：化可的松标准品色谱两见附录A中图A1.3.4.3案口试验
除不加试样多，均按上述测定驱过行。3.5结集的计第和常述
SN0700—1997
用色增教据处理机或按式(1)计算试中效化可的检费留含盘：A..V
式中：X一-试样中氢化可的松的残留含量，mg/kg，A——样疫中氢化可的松的色谱蜂面积,mmA.
标准工作绒中化可的愁的色谱峰面积+mm\，标准工作溶液中氢化可的松的浓度g/mL，V—样液册降定客体系，mT.
nt——-般终择所代表的试样量g。注，引算结费打除空白值，
4测定低限，回收率
4.1测定邮限
本方法的测定低浓为0.01mg/kg。4.2国临率
鲜牛乳中驾化可药松添加浓度及其国收率的密验数据：在0.01ag/kg时，同收率步.0%」在0.05mg/kg时.回收率为83.B%：在0.10mg/kg，回收汽次87.5%.
SV0700-1997
附录A
(示的路惑)
标准品色谱图
氛化可的松标准品的激相色谱图14
SN 07001997
Foreword
This sianlerl was irefted in aacardanoe with the requiretmente of GB/T 1. -1993\Directives fos.the work of standardization -Unit l, Drafing and presentation af atandarda-Part l, General rulefor drafting standerdg\ar.d SN/T 009l-1005\Gcncrel rulca for drufting lle lanedard mcthods for thedetcrminatior of pesticide, velerinary drug residues rd binsokina in cnmraodities fo: expurt\, Theme1had of ilalermiaatinll of chis standarc wae drafted by referring to reievant domestic and fareign lileratures through research ,modificalion and yerification, In addision, methods af samnpling and samplepreparation are also specified in this stedardThe limit af deterhinotion in this slundurd is delined on the. bhasia nf the curren. inlernaliurulma kimum limi1 lor hydrormrtisane zesiduen in milk and the sensitivity af the method.Annex A pf this standzrd is an informative aninex.Thig stendard was prapased by end ig ender the chergr of the Stile Acininislrulior of Import andExpurt Cutnmodity Inspnctian nf ihe Penple'Republic o: Chira.Thia etandard was dratted by Zhejiarg Inport and Export Cotnrnodity lespection Burezu of thePeople's Kepublic of China.
The nain draftcrs of this gtandlerd are Shung Xuebiu+ Zhuug Juu und SEsen Lisherng.This stauedard is a prclessjotal stanSN 0700—1997
This staadard apecifies the methode of sarupling+ sanple preparation und dererninatinn hy lighperforroanee liquid chrpmacug-uphy of hydrocortisone tekidnes in milk and mnilk products for export.This sratidlarl is applicahle to the deter mination ot hydrocortisone: residces lr. fresh milk for ex-porr.
2Samplingend samplegreparatlon2.1 Inspeetun lutwwW.bzxz.Net
The yuaminy ol aninspeseticn lnt should nptbemorethan 50 oo0bortlesThe characteristics of the cargo within the zame inspeetion Jor, guch &s peeking; Inerk, originspetification, grade etc. , ahouid be the sumc.2.2 Quanticy of aample tekenNumber of boteles ir?
ar: inepeetinn Ln.
L.ese thant 10 000
10 000:-20 000
20 G1—30 UO)
80 001-40 000
40 60:—50 000
2.3Sampling procedure
Minimium m:mberaE
bottles to be taken
A nurabar of bottles spocified in 2.2 ere tascll Et randon as priraary sample,The szmglec bttiesshouid be merkci nc. sent tu lnburuluty jr tine.2.dPrsuatinn nf leal anples
Mix the milk in the sampled bottles evenly and reduced tp cu 2so ul., wliclt is furelher livilaut inta two egual gortions, Each Fortinn in placcd in 2 clean coutaiaer ae che test aanp'e healerl anel in-heled.
2. 5 Srorage of lesl somple
Tiue lest samplea ahauld be storcd below 5' and kept swuy Froro ligh:Note, In thc course o: sempliny, rndl inmple penparutikan, grecauriona must he takun t avoid coutarize:ivo or nsfaztors wlich may erase the eivenge rf nylrcaurrisrme caalemt.Approved by theStateAdminigirtlonofEmpartndExportCommodity[nspectionafthe People's Kepublic of Chinu un Dec. 1.1SS76
Implemented Fruin Apr, 1: 19983 Melhod of determluatlon
3. Principle
5N0700-1997
The hydrocortisone residues ir, the test sample ure extrautel wih dichlorumethane and the organ-ie layer ig elesned up by washing with worljun hydroxite olution. The orgaic layer is then Eveporatcdta dryuess, and the resilue is dissolved in raobile phast, The reaultirg golution is extract with n-hex-ane to remove the fat, and finally analyzed by a high perlo:manee liquid ehromatograph with UV de.tectar. Exteinel stendard rethad ig ugcd for quentitatiwn3.2Reegents andmtcrials
Untess otherwise sperified, all the reagente used hall be analytically pure, \water\ig distilledwale:.
3.2.1 Methanol; HPLC grade.
3.2.2 Dichiorcmethenc:Redistill und colleet thu distillure sf 3g-41r.3.2.3n Hexane,Relistilled.
3. 2.4 Solium hydroxide solutian: 3. 1 mol/L agueous salution.3.2.5 Celive 45.
3.2.6Hydrorortisonc stendard:Purilyg,0%.3. 2. 7 Hydrurortinome glaadard saiuzian, Acaurately weigh an adequate rmount uf hydrucurtizaitieatandard. dissolve la mob:ls phase to prepare a stardard srpek sulutiun of 100 pg/mL in rancentra-tipr. Acoording to the requisemeut Freerc e stantkurd warkirig anli:tism af approgriate concentratiotby diluring the stock solutiun wil, nolilpnase.3.3Appsr:lus aarl eruipraent3.3.1 lEigli Ferlarmance liguid clromatograph, equigped with Uv deteezur3.3.2Rotary evaporator
3.3.3 Centrifugt.
3.3.4Veirirx miker.
3.3.5Air-flaw concenttating equpment3. 3. 6 Celive eolurar., 2h mrsx 25 inm (il>glan cn!urrn with scnponck, pecked with a small plzg afabaorbeal uotturl ul (lr h.ltoin, Ihen ith f. g, cf selite S45(3. 2. 5)3.3.7Separatoryfunrel:25o mL.3.3.B Cer.trifuge t:hens 5 ml.. witn stopprr.3. 3. 9 Micro-5yrirgc: 25 μL.3.4Prurdurr:
3. 4.1 Fxractinr: and -lnanrWeigb co10g of the tust snplekaceurrte lu C.22iatu a benker, neld ea 1cg ot celite 545,ardmis well, let send lur l0 min. Tle mixtare is trausferred iata the celite colunn (3. 3. 6). Ad 40 mLof dillniatuathane,end ter. stand for 1s tnin. Through a separetory fennel, drou 80 ml. frieh'nronuethano into tbe colemt: with s Jow rate (buch ir. and out frurt of 2-4 nl./min: Cnllect al)the effluents inco tnother aeperatury unnei: Waush The -fural wi l, 3) m]. w: godium hydroxice &olu.tian aad thea with S( rsJ. u: wa -i. rnd il:a.n thc organiz paane irte an evaposatsng flask. Rotury e-vapuril. u ta 2-3 ml. a: r. hnth temparature of 35. Transfer the concenzrated ao.usion into a re.Irifuye tebe. Riasc the watl ne f.ask with 2Xl mol. of diehloramcthare and corplie thr rinaings intathe ame centrifuge.tnbs. Corcenrate the soluticrilo rityness using a genil atream of eir. Add exactSN 07001997
ly l ml of mobile phase (seeurare to C, 02 mL) to the centrifuge tube: ard mix for ? rmin in a orexrixet, Extrarl thrce times witl l ml porions of r hexane. Por each time, vortex fur l rin, centrifuge at 3 Cno r/min fur 3 mir, pipet aut the w-hewarue lxyer and digcard. The aqucows layer is usedtfor Jiquid clurnmatogrophic determinalior.3. 4.2Determination
3. 4.2.7 LC opereting conditiana)ChromatogvaphicecolumarOIsCncolumn,12mmX4.0mm(id>,5pmh) Muhilr phas; Metaarml-water (45-55)+e) Flaw rste: 1.0 mL/min
d) Determinatio2 wave length: 245 nm:e) Colurmr. tenperature, Roor tempersture:I>lajcetianvo.um20pE.
3.4.2.2LCdetermimatiom
Accorriing to the aproximete conceatration oE bytirocortisole in the xrtrple wlutien, select ihestaadard workiag selution with similar peak atta to that ol the tamiple eolutioa. The rteponses of hy-drocortitane in the stardard wo:king solution and sample splurion ahould be in the linear range of theinstrumcntaj ictreioa, The stitndued werkirg solutian shaule! be rendomly inicetad in-betwecn -he in-jeutions aJ lh- sarnple solutinn af enual volume. TInder the sbove IC aprreting conditian, the reten-cina time af hydrnrtisone i6 sbout T. G min. For the liquid chrounatpgram ot the siaudard, aee fig. A)ia annex A.
3.4.3Blank tcst
The uperetiun uf the Llurk est is the sutre s that drkurioud in thr nelhod uf derrnuutioa, bulwilhoul udditian o[ sanyple.3.5Calculaticn and expression of the resultThe caleulation af che conient ol hydrpsortiscne residuas in the test sample ig caricd out by Ll.data procegaor, or areordir.g tu the fcrrnely (1):X-A..V
vrhere
X-Contrnt sf hydrxertisonc residreg ir. the lent gereple, Tg/kg:A-Przk are o hydrozoctisore in the sample salutiun, rrm;A,-Prelt aureu of bydruortisore in ttae gtandard wurking sululion,mn',c-Conccltration of hydrocortiaane in the stendard workirg sulution,ug/ml.V-Fimeil valune or il:e sruple soluliori,nn.Cn:respnindinr mags nf cte lexr sample in the Final sample soiution+ g-Nme: The ilank velun shnln hr sibtresind frnm inr nibewe roeult cf ra.ruiarion.E Limit of deterininmlion rund rerovery4. I Iimt of determinticnl
The iirrit f deterrtir:atiun al ili: teilul is o.ol ngkg+4.2Reavery
Acrrdirg to the exgerimental data tte tartifying cansentratiens of bydrosortieune in fresn milkand its corresponding recoveries ere:5
0. 01 mg/kg+the recovery 8a. 0%:0. 05 mg/kg,the recovery 83. 8%0. 10 mg/kg-the recovery 87. 0/6.SN 07001997
SN07001997
AunexA
(intormatjve)
Chramatgratn f the standard
wesgansaipky
Figure Al Liquid cmnatcgrzr of aydrccor tirot.e: ataniard
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