《畜禽肉制品中5種核苷酸的測定》編制說明研究背景我國是一個畜禽肉制品生產(chǎn)消費大國，隨著我國國民經(jīng)濟發(fā)展，消費者從關(guān)心食品安全到注重食品品質(zhì)的轉(zhuǎn)變。其中，核苷酸作為肉制品呈味物質(zhì)已被廣泛認同，游離核苷酸是已經(jīng)成為評判畜禽肉制品品質(zhì)及滋味的重要指標(biāo)。畜禽肉制品中，對肉制品味道產(chǎn)生影響主要作用的核苷酸包括腺苷酸（Adenosine5'-monophosphate，AMP）、肌苷酸（Inosinemonphosphate，IMP）、鳥苷酸（Guanosine?monophosphate，GMP）、肌苷（Hypoxanthineriboside，HxR）、次黃嘌呤（Hypoxanthine，Hx）5種，具體結(jié)構(gòu)如圖1所示。IMP、GMP、AMP本身就具有鮮味，并且會與谷氨酸鈉之間呈現(xiàn)強烈鮮味的相乘作用，Hx具有苦味，會給肉的整體風(fēng)味帶來負面作用，5種核苷酸之間組成和相互作用，形成畜禽肉制品的不同風(fēng)味。食品在加工過程中，細胞結(jié)構(gòu)被破壞，DNA暴露，在酶及物理條件作用下發(fā)生裂解，大部分三磷酸腺苷（Adenosine5'-Triphosphate，ATP）降解為AMP，進一步降解成IMP，IMP后續(xù)再次降解，產(chǎn)生HxR與Hx。在生產(chǎn)過程中，生產(chǎn)者根據(jù)產(chǎn)品工藝需要，也會人為添加核苷酸。在GB2760-2014中就規(guī)定IMP和GMP可在各類食品中按生產(chǎn)需要適量使用。HxHxHxHxHxRGMPIMPAMP圖15種核苷酸的結(jié)構(gòu)式HxRGMPIMPAMP目前，國際標(biāo)準化組織（ISO）對于核苷酸的關(guān)注集中在嬰兒配方奶粉中，已有BSISO20638《嬰兒配方奶粉液相色譜法測定核苷酸》，但是還沒有關(guān)于肉制品的核苷酸檢驗標(biāo)準。在我國僅有添加劑標(biāo)準GB1886.97-2015《食品安全國家標(biāo)準食品添加劑5'-肌苷酸二鈉》，沒有肉制品中核苷酸測定方法的國家標(biāo)準。在GB2760-2024中規(guī)定肌苷酸二鈉和鳥苷酸二鈉可在各類食品中按生產(chǎn)需要適量使用，但并沒有對應(yīng)檢測手段，國家層面的標(biāo)準更新滯后。近幾年，一些企業(yè)和團體為了適應(yīng)行業(yè)發(fā)展，也紛紛頒布團體標(biāo)準，其中畜禽肉品質(zhì)指標(biāo)的團體標(biāo)準TNAIA003-2020《肌肉中肌苷肌苷酸的測定高效液相色譜法》于2020年發(fā)布。本標(biāo)準能夠助力企業(yè)在產(chǎn)品研發(fā)時，對產(chǎn)品滋味的控制，促進企業(yè)產(chǎn)品升級，豐富產(chǎn)品品類。本標(biāo)準針對肉制品基質(zhì)，采用液相色譜法進行檢測。工作情況任務(wù)由來、起草單位、起草人根據(jù)《工業(yè)和信息化部辦公廳關(guān)于印發(fā)2023年第二批行業(yè)標(biāo)準制修訂和外文版項目計劃的通知》（工信廳科〔2023〕42號），由全國肉禽蛋制品標(biāo)準化技術(shù)委員會（SAC/TC399）負責(zé)《畜禽肉制品中5種核苷酸的測定》行業(yè)標(biāo)準（計劃編號為：2023-0950T-QB）的制定工作。中國肉類食品綜合研究中心承擔(dān)《禽肉制品中5種核苷酸的測定》的起草工作。主要起草人員為趙燕、郭文萍、趙文濤、任南、王真、紀曉萌。簡要起草過程2023年8月，我單位接到國家標(biāo)準修訂任務(wù)后，對該標(biāo)準的具體工作進行了認真研究，確定了總體工作方案，并組建了標(biāo)準修訂工作組，制定了實施計劃。2023年8月-12月，成立標(biāo)準編制小組。全面了解國內(nèi)外有關(guān)核苷酸的檢測方法以及標(biāo)準制定等情況，查閱國內(nèi)外相關(guān)參考文獻,了解目前核苷酸檢測的現(xiàn)狀，對相關(guān)企業(yè)及個人開展調(diào)查，完成實施方案的制定。2024年1月-7月，實驗研究。開展試驗研究，確定核苷酸測定方法的具體技術(shù)指標(biāo)。2024年8月1號，于中國肉類食品綜合研究中心院內(nèi)召開行業(yè)標(biāo)準起草工作組會議，起草組介紹標(biāo)準的相關(guān)內(nèi)容，與會專家針對標(biāo)準草案逐條討論，提出意見、建議。根據(jù)專家提出的修改意見，起草組對標(biāo)準文本進行修改。方法的起草原則在編制過程中，本起草組查閱了相關(guān)文獻報道，參考了各行業(yè)部門制定的同類標(biāo)準方法，注重標(biāo)準的可操作性，完成了檢測方法的研究工作，形成了方法草案和編制說明。本方法的主要參數(shù)、公式、性能要求等主要依據(jù)GB/T1.1-2009《標(biāo)準化工作指導(dǎo)》、GB/T20001.4-2015《標(biāo)準編寫規(guī)則》的要求進行編寫。并且按照GB/T27404-2008《實驗室質(zhì)量控制規(guī)范》對方法學(xué)進行了考察。標(biāo)準主要技術(shù)內(nèi)容本標(biāo)準研制過程中，通過查閱大量國內(nèi)外有關(guān)文獻，全面掌握了IMP、GMP、AMP、HxR、Hx的化學(xué)結(jié)構(gòu)和理化特性。在此基礎(chǔ)上，反復(fù)多次進行試驗，針對前處理方法、儀器條件、線性范圍、靈敏度和精確度等進行探索，并根據(jù)本實驗條件進行了多次試驗，最終確定了該方法的可行性，完成了檢測方法標(biāo)準的研究工作。儀器條件的優(yōu)化檢測波長的確定參考有關(guān)文獻，運用光電二級陣列管檢測器，對5種核苷酸進行掃描得到光譜圖2，發(fā)現(xiàn)HxR、IMP在248nm有最大吸收，GMP在254nm有最大吸收，AMP在259nm有最大吸收，Hx在250nm有最大吸收。將色譜圖疊加得到圖3，在254nm波長處，AMP、HxR光譜形成交叉點，Hx、IMP光譜形成交叉點，并且都具有較高的響應(yīng)，同時GMP在此波長下響應(yīng)值也較高，所以最終實驗確定選用檢測波長為254nm。在該波長下，5種核苷酸的響應(yīng)均可以滿足方法檢出限（limitofdetection，LOD）和定量限（limitofquantitation，LOQ）的要求。圖25種核苷酸的光譜圖圖3不同波長下5種核苷酸的響應(yīng)值色譜條件的確定選擇合適的色譜條件，保證5種核苷酸的色譜峰可以實現(xiàn)基線分離，是進行研究的第一步。在色譜柱和流動相選擇上，比較C18柱、HilicPlus柱、SB-Aq柱3種色譜柱，同時選取水/甲醇、甲酸水/甲醇、磷酸二氫鉀/甲醇、乙酸銨/甲醇的流動相體系，在等度洗脫條件下進行分離。通過查閱文獻可知，在5種核苷酸中，GMP和IMP較為難分離。結(jié)果發(fā)現(xiàn)，無論在何種流動相條件下，HilicPlus柱、SB-Aq柱均無法將GMP和IMP分離，圖4為HilicPlus柱所得的色譜圖，圖5為SB-Aq柱所得的色譜圖。C18柱在磷酸二氫鉀/甲醇流動相體系下，可以成功分離5種核苷酸。最終確定色譜柱為C18柱（250mm×4.6mm，5μm）；流動相：0.01mol/L磷酸二氫鉀∶甲醇＝98∶2（V/V）；流速：1.0mL/min；檢測波長：254nm；進樣量：20μL。HxR、IMP、GMP、AMP、Hx（10μg/mL）標(biāo)準溶液的色譜圖見圖6。IMPGMPIMPGMP圖4IMPGMPIMPGMP圖5SB-Aq柱所得的色譜圖GMPGMPIMPHxAMPHxR圖6C18柱所得的5種核苷酸標(biāo)準溶液色譜圖（10μg/mL）柱溫的確定調(diào)節(jié)柱溫箱的溫度，觀察不同柱溫下色譜峰的出峰時間和分離度的變化。分別考察了柱溫為25、30、35℃時的5種核苷酸的出峰情況，結(jié)果如圖7所示。結(jié)果表明，5種核苷酸的出峰時間在25及30℃條件下，出峰時間變化較小；35℃條件下，5種核苷酸的出峰時間后移，并且峰面積變??；不同溫度對于5種核苷酸的分離度影響較小。考慮到實驗室通常的儀器配置及檢測的環(huán)境溫度，因此選擇柱溫為25℃。ABCA.25℃；B.30℃；C.35℃圖7不同柱溫下5種核苷酸的出峰情況前處理條件的優(yōu)化提取溶劑的確定作為化合物分析的第1步，選擇合適的提取溶劑，將目標(biāo)物從樣品基質(zhì)中高效、快速地提取出來，直接影響方法的提取效率和后續(xù)的凈化步驟。本實驗選擇5%高氯酸＋5%甲醇水溶液、5%高氯酸、5%三氯乙酸、100%乙腈作為提取溶劑，分別考察4種溶劑的提取效果。圖8不同提取溶劑對加標(biāo)回收率的影響(n=3)對比加標(biāo)量為100mg/kg時的加標(biāo)回收率。由圖8可知，用100%乙腈溶液作提取溶劑時，5種核苷酸基本沒有回收，說明核苷酸很難溶解于有機溶劑，從而影響提取效率。用5%三氯乙酸溶液作提取溶劑時，目標(biāo)化合物的加標(biāo)回收率在62%左右。用5%高氯酸溶液和5%高氯酸＋5%甲醇水溶液作提取溶劑時，目標(biāo)化合物的加標(biāo)回收率在76%左右，但甲醇的加入會使實驗平行性變差，實驗偏差過大。因此選用高氯酸溶液作為提取溶劑。進一步對提取液的濃度進行優(yōu)化。選擇不同體積分數(shù)的高氯酸（1%、5%、10%、15%），分別考察其提取效果。對比加標(biāo)量為100mg/kg時的加標(biāo)回收率，實驗結(jié)果如圖9。結(jié)果表明，當(dāng)高氯酸的體積分數(shù)為1%時，回收率為50%左右；當(dāng)高氯酸體積分數(shù)為15%，回收率為63%左右，但結(jié)果偏差較大；當(dāng)高氯酸體積分數(shù)為5%、10%時，回收率約為74%。兩者回收率沒有明顯差異，考慮到酸會對后續(xù)處理產(chǎn)生影響，因此選擇體積分數(shù)較低的5%高氯酸溶液作為提取溶劑。圖9不同濃度的高氯酸對加標(biāo)回收率的影響(n=3)提取方式的確定畜禽肉制品中脂肪和蛋白質(zhì)的含量都很高，會對化合物的提取會有一定影響。選擇合適的提取方式，有助于減小基質(zhì)對于化合物的干擾，減少凈化難度。本實驗考察常見的4種提取方式：室溫超聲30min、冰浴超聲30min、常溫振蕩器振蕩30min、添加陶瓷均質(zhì)子渦旋30min。圖10不同提取方式對加標(biāo)回收率的影響(n=3)對比加標(biāo)量為100mg/kg時的加標(biāo)回收率，實驗結(jié)果如圖10。室溫超聲30min的回收率約為68%，冰浴超聲30min的回收率約為71%，兩者沒有明顯差異。常溫振蕩器振蕩30min的回收率約為80%，添加陶瓷均質(zhì)子渦旋30min的回收率約為77%，兩者在回收率上無明顯差異。但添加陶瓷均質(zhì)子渦旋30min得到的提取溶液表面有較多油脂懸浮，會對后續(xù)操作產(chǎn)生干擾，對比照片見圖11。因此在提取時選擇常溫振蕩器振蕩30min。圖11添加陶瓷均質(zhì)子渦旋30min（左）、常溫振蕩器振蕩30min（右）得到的提取溶液的狀態(tài)進一步對常溫振蕩器振蕩過程的提取時間進行優(yōu)化。選擇不同提取時間（10、20、30、40min），分別考察其提取效果。對比加標(biāo)量為100mg/kg時的加標(biāo)回收率，實驗結(jié)果如圖12。結(jié)果表明，在提取10min后，回收約為50%，且偏差較大，說明還未提取完全。隨著提取時間的延長，回收率逐漸升高，從30min到40min時，回收率逐漸穩(wěn)定在80%左右。因此選擇提取時間為30min。ABCDEA.AMP；B.IMP；C.GMP；D.HxR；E.Hx圖12不同提取時間對加標(biāo)回收率的影響(n=3)離心轉(zhuǎn)速的選擇在樣品振蕩提取后進行離心，可以將樣品中不溶物沉降，實現(xiàn)提取液的固液分離，方便進行下一步操作。離心轉(zhuǎn)速是該過程的核心參數(shù)。本實驗考察6000、8000、12000、15000r/min的轉(zhuǎn)速，觀察其離心效果。實驗結(jié)果如圖13所示。在6000及8000r/min條件下，上清液仍舊有少量固體物質(zhì)殘留，上層溶液比較渾濁；在12000及15000r/min條件下，上清液明顯變澄清，說明不溶物已經(jīng)沉降完全，且12000及15000r/min的狀態(tài)大致相同。因此，實驗中選擇離心轉(zhuǎn)速為12000r/min。6000r/min6000r/min8000r/min12000r/min15000r/min圖13不同離心機轉(zhuǎn)速的沉降效果比較提取液最終pH的確定在提取過程中，需要用到大量的酸來保證游離核苷酸充分離子化，充分溶解。但提取液中酸會對后續(xù)操作產(chǎn)生干擾，也會影響色譜柱柱效，影響分析物保留時間和分離效果。因此，在定容之前需要對提取液的pH進行調(diào)整。本實驗比較不同pH值提取溶劑（5.5、6.0、6.5、7.0）的提取效果。ABCDEA.AMP；B.IMP；C.GMP；D.HxR；E.Hx圖14提取液最終pH對加標(biāo)回收率的影響(n=3)對比加標(biāo)量為100mg/kg時的加標(biāo)回收率，實驗結(jié)果如圖14。提取溶液pH從5.5到6.5，回收率隨pH的升高而升高，提取溶液pH從6.5到7.0，回收率隨pH的升高而降低，在pH6.5時回收率最高。因此選擇提取液pH為6.5。為了進一步鎖定合適的pH范圍，對pH進一步細化。本實驗設(shè)置了幾個不同的提取液pH（6.1、6.3、6.5、6.7、6.9），探究了其對回收率的影響。實驗結(jié)果如圖15。取溶液pH從6.1到6.3，回收率隨pH的升高而升高，pH從6.3到6.7，回收率隨pH的變化并無明顯區(qū)別，提取溶液pH從6.7到6.9，回收率隨pH的升高而降低。因此，確定提取液pH為6.5±0.2。ABCDEA.AMP；B.IMP；C.GMP；D.HxR；E.Hx圖15提取液最終pH范圍對加標(biāo)回收率的影響(n=3)提取次數(shù)的確定從之前的優(yōu)化結(jié)果看，最優(yōu)回收率約為80%，回收率仍舊偏低，為了提高回收率，考慮通過增加提取次數(shù)的方式來提高回收率。實驗結(jié)果如圖16。提取一次的回收率為83%，提取兩次的回收率為90%，提取三次的回收率為91%，提取兩次和提取三次的回收率沒有明顯差異。因此，實驗采用兩次提取方式，實驗最終回收率達到91%。圖16提取次數(shù)對加標(biāo)回收率的影響(n=3)凈化方式的確定樣品經(jīng)過提取后，上清液中還含有大量油脂和蛋白質(zhì)等雜質(zhì)，雜質(zhì)的分子質(zhì)量較大，在流動相中流動緩慢，極易堵塞色譜柱，還會對目標(biāo)物的檢測產(chǎn)生基質(zhì)干擾，降低方法的靈敏度。為了能有效對樣品進行凈化，除去油脂、蛋白質(zhì)等基質(zhì)干擾，需要選擇合適的凈化方式。實驗中選擇了固相萃取柱法、液液萃取法2種方式進行樣品凈化。經(jīng)查閱文獻，選擇了MCX柱、HLB柱、WAX柱、PSA柱等不同固相萃取柱類型。實驗發(fā)現(xiàn)，由于5種核苷酸的化學(xué)性質(zhì)差異較大，沒有一種固相萃取柱可以對5種核苷酸均有較好保留。使用通過型PRIMEHLB固相萃取柱，發(fā)現(xiàn)過柱后的提取液并未變得澄清，色譜圖中雜質(zhì)峰也未減少，而且會導(dǎo)致樣品平行性變差。后采用正己烷液液萃取法除去雜質(zhì)，經(jīng)正己烷凈化后的提取溶液變更澄清，實驗結(jié)果見圖17，同時核苷酸的回收率沒有減小。因此，凈化方式采用正己烷液液萃取法。未除脂的提取液除脂后的提取液未除脂的提取液除脂后的提取液圖17經(jīng)正己烷液液萃取法除雜質(zhì)前后提取液狀態(tài)的比較方法學(xué)考察標(biāo)準曲線、檢出限及定量限標(biāo)準儲備液（1mg/mL）：分別精密稱定HxR、IMP、GMP、AMP、Hx約25mg（精確至0.00001g），用水溶解后，定容至25mL容量瓶中。配制成濃度均為1mg/mL的標(biāo)準儲備液，于4℃保存，有效期3個月。混合標(biāo)準工作液（100μg/mL）：分別準確移取各標(biāo)準儲備液5mL，用水稀釋并定容于50mL容量瓶中，配制成濃度均為100μg/mL的標(biāo)準工作液，于4℃保存，有效期1個月。混合標(biāo)準系列工作溶液：準確吸取混合標(biāo)準工作液0.0500、0.500、2.00、5.00、8.00mL至10mL容量瓶，用水定容，配制成質(zhì)量濃度為0.500、5.00、20.0、50.0、80.0μg/mL的混合標(biāo)準系列工作溶液，臨用現(xiàn)配。用優(yōu)化好的液相色譜條件測定系列標(biāo)準工作液。以測得峰面積（Y）為縱坐標(biāo)，對應(yīng)的標(biāo)準溶液濃度（X）為橫坐標(biāo)，繪制標(biāo)準曲線。其回歸方程和決定系數(shù)如表1。5種核苷酸在0.5～80μg/mL濃度范圍內(nèi)呈現(xiàn)良好的線性關(guān)系。表15種核苷酸的回歸方程和相關(guān)系數(shù)物質(zhì)線性范圍線性方程決定系數(shù)R2AMP0.5～80μg/mLY=0.1867X+0.00850.9998GMPY=0.1585X+0.00140.9997IMPY=0.1585X-0.00030.9999HxRY=0.2298X-0.00270.9999HxY=0.3580X+0.00740.9996本方法選取了豬肉脯、醬牛肉、臘肉、火腿腸、燒雞作為代表樣品，添加適量標(biāo)準工作液進行試驗，考察方法的檢出限和定量限。經(jīng)回收率試驗，當(dāng)添加量為5mg/kg時，信噪比（RS/N）均大于3；當(dāng)添加量為15mg/kg時，信噪比（RS/N）均大于10，色譜圖見圖18。所以本方法的檢出限確定為5.0mg/kg，定量限確定為15.0mg/kg。ABA.檢出限；B.定量限圖18檢出限及定量限的色譜圖穩(wěn)定性考察按照設(shè)計好的時間，將配制的5種核苷酸的儲備液于4℃冰箱保存，根據(jù)1個月、2個月、3個月、4個月、5個月、6個月的保存時間進行取樣并分析，記錄化合物峰面積。按以下公式計算剩余分析物（%）。剩余分析物（%）=Ci/C0×100%。其中：Ci=不同時間點的響應(yīng)值；C0=新配標(biāo)準儲備溶液的第一次測得的響應(yīng)值。根據(jù)表2的數(shù)據(jù)變化，儲備液在三個月之內(nèi)，響應(yīng)值變化較小，結(jié)果表明，標(biāo)準儲備液有效期定為3個月是可行的。表2標(biāo)準儲備液剩余分析物的變化物質(zhì)1個月2個月3個月4個月5個月6個月AMP99.599.298.396.591.286.7GMP99.799.198.595.691.688.5IMP99.399.299.096.390.283.6HxR99.298.998.695.692.185.6Hx99.998.598.896.191.181.9將儲備液稀釋成濃度為100μg/mL的混合標(biāo)準工作液，并于4℃冰箱保存，按照0.5個月、1個月、1.5個月、2個月的保存時間進行取樣并分析，記錄化合物峰面積。按以下公式計算剩余分析物（%）。剩余分析物（%）=Ci/C0×100%。其中：Ci=不同時間點的響應(yīng)值；C0=新配標(biāo)準儲備溶液的第一次測得的響應(yīng)值。根據(jù)表3的數(shù)據(jù)變化，混合標(biāo)準工作液在1個月之內(nèi)，響應(yīng)值變化較小。因此，混合標(biāo)準工作液有效期定為1個月是可行的。表3標(biāo)準工作液剩余分析物的變化物質(zhì)0.5個月1個月1.5個月2個月AMP99.698.195.688.6GMP99.898.895.789.2IMP99.798.293.890.3HxR99.598.793.190.1Hx99.998.995.789.7方法的回收率按照GB/T19480-2025《肉與肉制品術(shù)語》中對肉制品的分類，選取不同加工工藝以及居民消費頻率，本實驗選取了豬肉脯、醬牛肉、臘肉、火腿腸、燒雞作為代表樣品，在樣品中添加15mg/kg、30mg/kg、150mg/kg（LOQ、2LOQ、10LOQ）三個不同濃度的核苷酸進行回收率試驗，每個濃度進行6個平行試驗，重復(fù)進行3次，實驗色譜圖見圖19-23，數(shù)據(jù)結(jié)果見表4-8。在本實驗室內(nèi)，15mg/kg～150mg/kg添加濃度范圍的回收率為80.0%-109.5%。批內(nèi)相對標(biāo)準偏差≤9.8%，批間相對標(biāo)準偏差≤7.7%。豬肉脯基質(zhì)豬肉脯基質(zhì)豬肉脯基質(zhì)加標(biāo)質(zhì)加標(biāo)圖19豬肉脯基質(zhì)及基質(zhì)加標(biāo)的色譜圖醬牛肉基質(zhì)醬牛肉基質(zhì)醬牛肉基質(zhì)加標(biāo)圖20醬牛肉基質(zhì)及基質(zhì)加標(biāo)的色譜圖臘肉基質(zhì)加標(biāo)臘肉基質(zhì)加標(biāo)臘肉基質(zhì)加標(biāo)圖21臘肉基質(zhì)及基質(zhì)加標(biāo)的色譜圖火腿腸基質(zhì)火腿腸基質(zhì)火腿腸基質(zhì)加標(biāo)圖22火腿腸基質(zhì)及基質(zhì)加標(biāo)的色譜圖燒雞基質(zhì)加標(biāo)燒雞基質(zhì)加標(biāo)燒雞基質(zhì)加標(biāo)圖23燒雞基質(zhì)及基質(zhì)加標(biāo)的色譜圖

表4實驗室內(nèi)肌苷回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ93.995.988.180.690.691.090.05.990.35.1Ⅱ85.381.392.789.481.685.185.95.2Ⅲ84.5104.0103.493.096.888.695.18.330Ⅰ93.694.9100.486.399.794.895.05.393.92.5Ⅱ88.685.193.8103.180.995.891.28.8Ⅲ104.6102.996.388.098.183.795.68.6150Ⅰ99.4100.794.992.896.796.796.93.093.13.7Ⅱ94.385.291.790.986.891.390.03.8Ⅲ95.193.994.183.099.588.692.46.2醬牛肉15Ⅰ84.196.490.099.182.890.490.57.293.63.2Ⅱ89.192.3103.896.5102.394.096.36.0Ⅲ96.684.597.599.495.291.194.15.830Ⅰ90.9101.295.4101.695.491.896.14.795.03.9Ⅱ94.7101.686.487.693.581.190.88.0Ⅲ100.997.394.791.7103.5100.398.14.4150Ⅰ89.499.587.497.497.199.195.05.593.53.9Ⅱ90.899.286.297.6101.9101.396.26.6Ⅲ97.882.888.092.890.484.789.46.1臘肉15Ⅰ98.888.584.7103.298.685.093.18.696.63.3Ⅱ104.697.9102.894.990.492.497.25.8Ⅲ97.2105.1100.7103.294.995.399.44.330Ⅰ95.1104.196.9100.0105.391.798.95.397.02.0Ⅱ96.4102.791.191.6102.199.597.25.2Ⅲ97.698.990.096.196.390.594.93.9150Ⅰ91.089.787.396.8100.294.793.35.294.53.1Ⅱ100.2104.6100.790.194.896.797.95.2Ⅲ84.592.1104.189.296.488.492.57.5火腿腸15Ⅰ93.691.290.089.8101.7103.995.06.593.21.9Ⅱ92.885.390.792.794.693.291.63.6Ⅲ95.187.493.088.696.996.392.94.330Ⅰ87.391.993.486.194.195.291.34.192.34.1Ⅱ94.886.487.184.993.487.589.04.6Ⅲ104.0104.496.092.091.091.496.56.5150Ⅰ99.282.6103.697.1106.598.898.08.596.81.5Ⅱ86.4101.791.887.9105.698.095.28.1Ⅲ90.088.0105.7104.491.3103.697.28.4燒雞15Ⅰ103.7105.498.384.192.7102.597.88.395.85.1Ⅱ83.480.293.391.696.596.590.37.6Ⅲ97.2108.195.392.6108.795.099.57.130Ⅰ105.284.287.888.091.693.691.78.095.83.8Ⅱ104.2102.8103.497.1100.285.298.87.2Ⅲ103.489.6101.291.798.795.796.75.6150Ⅰ91.489.8104.395.6105.694.596.96.893.03.7Ⅱ101.592.394.990.686.785.691.96.3Ⅲ96.993.388.795.981.385.690.36.8表5實驗室內(nèi)肌苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ97.186.785.184.892.188.489.05.495.15.6Ⅱ101.1100.396.6105.193.595.898.74.3Ⅲ101.597.895.294.8104.792.097.74.830Ⅰ101.391.589.298.583.7102.894.58.092.62.6Ⅱ98.096.195.698.080.291.893.37.3Ⅲ80.884.696.691.490.995.289.96.8150Ⅰ102.2100.294.681.6101.598.096.48.095.43.6Ⅱ90.583.888.097.392.997.491.75.8Ⅲ96.093.298.5100.299.5102.298.33.3醬牛肉15Ⅰ93.882.692.097.496.590.792.25.894.13.4Ⅱ84.695.9105.2103.696.4101.297.87.6Ⅲ98.694.389.291.085.496.092.45.230Ⅰ103.697.397.495.095.995.497.43.396.16.3Ⅱ83.980.896.686.999.089.689.58.0Ⅲ101.999.4102.1101.5105.397.7101.32.6150Ⅰ96.2101.196.796.395.798.597.42.195.53.1Ⅱ97.395.8103.795.398.790.997.04.4Ⅲ89.190.398.790.197.487.392.25.1臘肉15Ⅰ99.284.095.881.985.792.889.97.890.73.3Ⅱ88.796.991.195.798.193.394.03.8Ⅲ87.792.190.493.085.181.088.25.230Ⅰ102.292.597.2103.6102.584.797.17.694.04.1Ⅱ90.299.697.490.5102.890.395.15.8Ⅲ96.299.582.686.887.884.789.67.5150Ⅰ103.0103.197.495.298.593.098.44.294.64.2Ⅱ96.180.893.394.290.488.190.56.1Ⅲ86.083.0102.999.599.898.394.98.7火腿腸15Ⅰ99.386.296.590.8104.097.795.86.695.12.2Ⅱ91.994.8109.595.094.395.396.86.6Ⅲ94.390.991.094.8102.184.092.96.430Ⅰ89.396.490.487.391.293.991.43.691.50.9Ⅱ93.099.590.287.089.794.692.34.8Ⅲ98.089.291.088.487.690.590.84.1150Ⅰ98.990.390.087.390.1104.993.67.394.03.8Ⅱ105.898.2100.390.986.9104.997.87.7Ⅲ91.887.697.386.189.891.390.74.3燒雞15Ⅰ97.9100.1100.696.888.386.895.16.392.02.9Ⅱ99.283.6100.783.382.196.991.09.7Ⅲ92.383.787.097.094.585.790.05.930Ⅰ103.592.799.4103.288.683.095.18.892.85.6Ⅱ104.890.993.298.386.7105.096.57.8Ⅲ84.583.192.980.494.885.686.96.6150Ⅰ97.588.689.6102.683.488.291.77.792.52.3Ⅱ94.097.6100.790.891.794.494.93.9Ⅲ87.487.985.1103.191.390.290.87.0表6實驗室內(nèi)鳥苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ97.485.081.186.190.888.088.16.491.43.6Ⅱ98.686.098.886.691.588.291.66.3Ⅲ91.290.193.296.3102.794.194.64.830Ⅰ87.792.599.490.391.597.693.24.894.31.4Ⅱ80.996.998.293.096.199.594.17.3Ⅲ98.797.795.5102.882.397.495.77.3150Ⅰ97.992.793.299.397.792.995.63.190.74.7Ⅱ93.490.881.482.481.697.387.87.9Ⅲ88.997.483.384.785.792.788.86.1醬牛肉15Ⅰ83.687.690.395.891.490.489.94.591.34.1Ⅱ80.283.399.588.486.193.888.68.0Ⅲ90.296.085.5103.2103.195.195.57.330Ⅰ85.990.795.890.387.698.691.55.393.22.8Ⅱ101.7101.493.196.289.395.696.25.0Ⅲ97.393.097.381.796.985.692.07.3150Ⅰ90.294.196.793.694.886.592.74.093.83.3Ⅱ92.483.489.3103.284.196.091.48.2Ⅲ103.485.8100.2103.598.193.197.47.0臘肉15Ⅰ82.991.790.294.091.189.990.04.294.34.3Ⅱ92.9108.099.099.792.994.997.95.9Ⅲ103.590.394.791.690.2100.095.15.830Ⅰ91.295.595.896.997.595.295.42.391.24.3Ⅱ80.484.881.593.297.388.987.77.6Ⅲ94.388.083.4101.986.089.090.47.4150Ⅰ81.096.785.897.786.188.289.37.492.74.0Ⅱ88.195.996.093.9104.3101.096.55.8Ⅲ92.491.790.7102.987.288.592.26.1火腿腸15Ⅰ103.495.099.590.298.097.397.24.694.22.8Ⅱ88.696.6103.188.387.592.092.76.6Ⅲ87.386.499.097.594.191.392.65.630Ⅰ97.195.591.093.2104.894.396.05.093.72.2Ⅱ103.396.294.380.386.598.293.19.0Ⅲ80.584.798.689.998.599.792.08.9150Ⅰ102.492.492.7102.386.883.393.38.491.02.6Ⅱ84.1102.480.090.098.292.091.19.2Ⅲ100.496.584.583.685.580.888.68.9燒雞15Ⅰ84.488.584.590.294.298.490.06.192.12.1Ⅱ103.790.090.887.795.787.992.66.6Ⅲ95.993.487.297.198.790.093.74.730Ⅰ85.185.7101.188.096.091.591.26.993.62.5Ⅱ85.592.197.295.996.394.493.64.6Ⅲ86.798.597.490.998.2104.396.06.5150Ⅰ86.882.697.186.793.985.788.86.289.93.6Ⅱ84.392.597.087.782.280.687.47.3Ⅲ95.290.291.793.993.397.193.62.6表7實驗室內(nèi)腺苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ97.186.785.184.882.188.487.46.089.33.4Ⅱ81.1100.396.685.197.595.892.78.3Ⅲ81.587.895.284.884.792.087.75.830Ⅰ81.391.589.288.583.782.886.24.891.25.0Ⅱ98.096.195.698.090.291.895.03.4Ⅲ100.889.696.681.490.995.292.47.3150Ⅰ82.290.294.681.691.598.089.77.491.05.6Ⅱ80.583.888.087.392.987.486.74.8Ⅲ96.093.298.590.299.5102.296.64.5醬牛肉15Ⅰ83.892.692.087.486.580.787.25.392.55.8Ⅱ94.695.995.2103.696.4101.297.83.8Ⅲ98.694.389.291.085.496.092.45.230Ⅰ103.687.387.495.095.995.494.16.594.16.7Ⅱ83.980.896.686.989.089.687.86.2Ⅲ101.999.4102.1101.5100.397.7100.51.7150Ⅰ96.2101.196.796.395.786.595.45.094.32.0Ⅱ97.395.8103.795.388.790.995.35.5Ⅲ89.190.398.790.197.487.392.25.1臘肉15Ⅰ99.2104.095.891.995.792.896.64.691.94.9Ⅱ88.786.981.195.780.193.387.67.2Ⅲ87.792.190.493.085.1101.091.66.030Ⅰ102.292.597.298.6102.594.798.04.193.75.5Ⅱ90.299.697.490.5102.890.395.15.8Ⅲ96.289.582.686.887.884.787.95.4150Ⅰ83.0103.197.485.290.593.092.08.294.13.8Ⅱ96.180.893.394.290.498.192.26.7Ⅲ86.0103.0102.999.599.898.398.36.4火腿腸15Ⅰ89.386.296.590.884.087.789.14.992.43.1Ⅱ91.994.889.595.094.395.393.52.5Ⅲ94.390.991.094.8102.194.094.54.330Ⅰ89.386.490.487.391.283.988.13.191.53.4Ⅱ83.099.5100.287.089.794.692.37.5Ⅲ98.089.291.088.497.6100.594.15.5150Ⅰ88.990.390.087.390.1104.991.97.092.42.1Ⅱ95.898.290.390.986.9104.994.56.9Ⅲ81.897.697.386.189.891.390.76.8燒雞15Ⅰ97.9100.180.686.888.386.890.18.290.92.4Ⅱ99.283.690.783.382.196.989.38.4Ⅲ92.383.787.097.094.5105.793.48.330Ⅰ83.592.799.4103.288.683.091.79.191.73.4Ⅱ94.890.993.298.386.7105.094.86.7Ⅲ94.583.192.980.494.885.688.67.1150Ⅰ97.588.689.6102.683.488.291.77.792.45.0Ⅱ84.097.6100.780.881.784.488.29.8Ⅲ97.497.995.1103.191.399.297.34.1表8實驗室內(nèi)次黃嘌呤回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ97.196.785.184.8102.198.494.07.891.67.7Ⅱ81.180.386.685.183.585.883.73.1Ⅲ91.597.895.294.8104.799.097.24.730Ⅰ91.381.589.288.583.792.887.85.091.53.9Ⅱ98.096.195.698.090.291.895.03.4Ⅲ100.884.696.681.490.995.291.68.1150Ⅰ92.290.294.681.691.598.091.46.093.63.9Ⅱ90.583.888.097.392.997.491.75.8Ⅲ96.093.298.597.299.5102.297.83.2醬牛肉15Ⅰ83.882.692.087.486.590.787.24.291.94.9Ⅱ94.695.995.293.696.4101.296.22.8Ⅲ98.694.389.291.085.496.092.45.230Ⅰ93.697.397.495.095.995.495.81.591.65.4Ⅱ83.980.886.686.989.089.686.13.8Ⅲ101.999.492.181.585.397.793.08.8150Ⅰ96.2101.196.796.395.798.597.42.193.34.6Ⅱ97.395.883.795.398.790.993.65.9Ⅲ89.190.388.790.187.487.388.81.4臘肉15Ⅰ99.294.095.881.985.792.891.67.190.85.0Ⅱ88.786.991.185.780.183.386.04.5Ⅲ87.792.1100.493.095.1101.094.95.430Ⅰ102.292.597.293.692.594.795.53.990.65.1Ⅱ80.299.697.490.592.880.390.19.2Ⅲ86.289.582.686.887.884.786.32.8150Ⅰ103.093.197.495.290.593.095.44.694.74.1Ⅱ86.180.893.394.290.498.190.56.9Ⅲ96.093.0102.999.599.898.398.33.5火腿腸15Ⅰ99.386.296.590.884.087.790.86.691.22.2Ⅱ91.994.889.595.094.395.393.52.5Ⅲ94.390.981.094.892.184.089.56.430Ⅰ89.396.490.487.391.293.991.43.689.82.3Ⅱ83.099.590.287.089.794.690.76.4Ⅲ88.089.291.088.487.680.587.54.1150Ⅰ98.990.390.087.390.184.990.35.291.21.5Ⅱ85.898.2100.390.986.994.992.86.4Ⅲ91.887.697.386.189.891.390.74.3燒雞15Ⅰ97.9100.1100.696.888.386.895.16.392.02.9Ⅱ99.283.6100.783.382.196.991.09.7Ⅲ92.383.787.097.094.585.790.05.930Ⅰ103.592.799.493.288.683.093.47.990.03.2Ⅱ84.880.993.298.386.785.088.27.3Ⅲ94.583.192.980.494.885.688.67.1150Ⅰ97.588.689.692.683.488.290.05.390.84.1Ⅱ94.097.6100.790.891.794.494.93.9Ⅲ87.487.985.183.191.390.287.53.5實驗室內(nèi)的結(jié)果驗證試驗實驗室內(nèi)部組織雙人雙儀器進行方法學(xué)驗證，以豬肉脯、醬牛肉、臘肉、火腿腸、燒雞作為代表樣品，在樣品中添加15mg/kg、30mg/kg、150mg/kg（LOQ、2LOQ、10LOQ）三個不同濃度的核苷酸進行回收率試驗，每個濃度進行6個平行試驗，數(shù)據(jù)結(jié)果見表9。實驗室內(nèi)部組織雙人雙儀器，15mg/kg～150mg/kg添加濃度范圍的平均回收率為85.4%-101.8%。批內(nèi)相對標(biāo)準偏差≤9.6%，批間相對標(biāo)準偏差≤8.1%。表9實驗室雙人雙儀器驗證實驗結(jié)果待測組織添加濃度(mg/kg)人員平均回收率（%）RSD(%)平均回收率(%)雙人雙儀器RSD(%)豬肉脯15人員194.07.891.67.7人員283.73.130人員187.85.091.53.9人員295.03.493.63.9150人員191.68.1人員291.46.0醬牛肉15人員191.75.891.94.9人員297.83.230人員187.24.2人員296.22.891.65.4150人員192.45.2人員295.81.5臘肉15人員186.13.893.34.6人員293.08.830人員197.42.1人員293.65.990.85.0150人員188.81.4人員291.67.1火腿腸15人員186.04.590.65.1人員294.95.430人員195.53.9人員290.19.294.74.1150人員186.32.8人員295.44.6燒雞15人員190.56.991.22.2人員298.33.530人員190.86.6人員293.52.589.82.3150人員189.56.4人員291.43.6

實驗室間的結(jié)果驗證試驗方法標(biāo)準制定完成后，標(biāo)準編制小組委托山東省食品藥品檢驗研究院、浙江工商大學(xué)海洋食品研究院、中國海關(guān)科學(xué)技術(shù)研究中心三家單位對方法標(biāo)準的靈敏度、準確度、精密度和不同基質(zhì)的適用性進行了驗證，驗證結(jié)論為該方法可操作性強，準確度高、重復(fù)性好，滿足藥殘檢測的要求。各單位的驗證結(jié)果及實驗室間的重復(fù)性結(jié)果見表10～表24，驗證報告掃描件見附錄A。

表10山東院肌苷回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ101.3100.096.094.787.381.393.48.394.32.0Ⅱ90.7100.792.784.792.097.393.06.0Ⅲ103.3101.3102.788.786.796.096.47.630Ⅰ94.081.786.089.793.098.090.46.592.21.7Ⅱ93.387.795.089.791.3102.793.35.7Ⅲ103.089.386.098.395.385.792.97.6150Ⅰ93.389.185.383.581.889.687.14.992.68.7Ⅱ102.1106.6104.392.2105.1100.6101.85.1Ⅲ82.990.092.794.890.082.288.85.8醬牛肉15Ⅰ92.798.096.084.086.094.091.86.190.12.5Ⅱ84.084.784.081.398.093.387.67.5Ⅲ85.393.391.3100.786.788.791.06.130Ⅰ102.795.390.094.392.098.095.44.792.13.2Ⅱ86.793.390.784.0100.785.390.16.9Ⅲ93.096.395.084.791.084.090.75.8150Ⅰ91.5101.5105.889.8102.197.598.06.494.63.6Ⅱ104.991.790.792.592.395.194.55.6Ⅲ89.281.492.5102.390.292.091.37.4臘肉15Ⅰ94.792.081.384.790.786.788.35.689.22.8Ⅱ90.090.083.392.785.382.087.24.9Ⅲ90.089.3102.094.788.088.092.06.030Ⅰ98.384.393.091.094.084.790.96.190.12.1Ⅱ94.783.798.0100.388.783.391.48.0Ⅲ94.091.380.787.788.385.387.95.3150Ⅰ96.289.495.282.699.592.392.56.490.32.2Ⅱ87.988.287.585.9100.782.088.77.1Ⅲ93.389.398.586.385.585.489.75.9火腿腸15Ⅰ94.086.787.394.095.386.790.74.692.62.1Ⅱ95.385.389.399.399.398.794.66.3Ⅲ93.3103.384.092.791.390.792.66.730Ⅰ101.397.390.087.391.395.093.75.590.45.2Ⅱ90.095.398.788.397.785.392.65.9Ⅲ80.384.786.086.086.786.385.02.8150Ⅰ95.196.384.6102.197.0105.196.77.395.12.6Ⅱ93.986.788.185.1102.397.392.27.3Ⅲ93.291.395.6106.193.998.496.45.5燒雞15Ⅰ91.382.796.083.384.098.789.37.890.14.1Ⅱ84.083.396.782.088.087.386.96.1Ⅲ96.086.7103.386.799.393.394.27.230Ⅰ96.791.7103.089.796.386.794.06.292.91.1Ⅱ97.382.0100.789.398.088.392.67.8Ⅲ91.795.388.096.780.7100.392.17.6150Ⅰ106.2102.7103.593.698.3103.2101.24.596.24.6Ⅱ91.793.294.988.999.789.793.04.2Ⅲ90.096.792.498.295.193.194.33.2表11山東院肌苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ93.386.788.092.094.791.391.03.493.63.1Ⅱ88.797.3102.7100.0101.390.796.86.0Ⅲ92.794.788.794.793.394.093.02.430Ⅰ95.391.389.080.380.384.086.77.191.44.8Ⅱ85.097.795.094.797.3102.795.46.1Ⅲ89.0102.395.792.782.390.792.17.3150Ⅰ95.499.9102.1105.3102.7105.5101.83.7100.81.9Ⅱ98.398.7102.996.4100.394.998.62.9Ⅲ103.197.2106.097.0106.7101.5101.94.1醬牛肉15Ⅰ90.098.0103.398.7102.094.797.85.096.51.2Ⅱ93.395.399.392.096.796.095.42.7Ⅲ101.3102.797.388.794.792.796.25.530Ⅰ102.780.791.096.7102.084.392.99.992.90.1Ⅱ83.387.085.7103.397.0101.392.99.3Ⅲ102.392.783.794.395.089.092.86.7150Ⅰ96.195.9105.098.7101.594.498.64.198.40.4Ⅱ101.895.3102.499.198.894.998.73.2Ⅲ99.395.998.1102.795.096.798.02.9臘肉15Ⅰ86.790.088.098.086.7100.791.76.795.33.8Ⅱ103.399.3101.3103.386.0100.098.96.6Ⅲ101.390.797.393.398.092.095.44.330Ⅰ100.795.792.093.098.083.093.76.692.92.6Ⅱ87.780.783.088.3102.399.390.29.7Ⅲ99.089.091.097.789.7102.394.85.9150Ⅰ98.7100.094.598.395.193.996.82.698.83.3Ⅱ94.795.598.194.4103.396.597.13.4Ⅲ105.0103.795.6106.4102.1102.9102.63.7火腿腸15Ⅰ100.096.799.394.094.097.396.92.694.22.5Ⅱ101.398.790.392.087.387.392.86.4Ⅲ96.090.788.096.097.389.392.94.330Ⅰ99.394.093.099.390.381.392.97.292.02.8Ⅱ94.085.398.780.386.390.089.17.4Ⅲ89.795.087.390.7100.3101.394.16.2150Ⅰ95.996.3105.1103.0106.3106.6102.24.8101.30.8Ⅱ99.996.1105.9105.995.1101.6100.74.6Ⅲ100.795.3104.1106.5103.195.4100.84.6燒雞15Ⅰ87.386.786.090.790.092.088.82.791.53.0Ⅱ88.786.789.389.3100.794.091.45.6Ⅲ94.798.794.093.398.087.394.34.330Ⅰ100.087.087.084.380.092.088.47.891.32.8Ⅱ99.387.380.086.3101.0100.792.49.8Ⅲ84.0103.097.795.780.798.093.29.4150Ⅰ95.996.994.797.794.994.695.81.499.63.4Ⅱ94.2102.495.4104.8106.3100.5100.64.9Ⅲ105.598.1106.1105.7100.598.7102.43.7表12山東院鳥苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ103.396.0102.794.097.396.098.23.994.63.6Ⅱ88.799.3100.794.787.393.394.05.8Ⅲ91.393.394.094.788.087.391.43.430Ⅰ103.397.0101.7102.3100.798.7100.62.499.01.7Ⅱ102.798.7101.094.3101.396.799.13.2Ⅲ99.3103.395.395.796.093.797.23.6150Ⅰ104.9101.5105.397.198.395.9100.54.099.71.4Ⅱ94.3101.0100.799.495.398.198.12.9Ⅲ104.998.299.896.4104.998.7100.53.6醬牛肉15Ⅰ88.090.0102.0100.795.388.794.16.696.23.2Ⅱ103.3100.7101.3100.7102.090.799.84.6Ⅲ98.793.387.3100.786.7101.394.76.930Ⅰ100.093.7101.398.096.394.097.23.298.21.0Ⅱ101.798.397.794.395.0102.098.23.3Ⅲ97.396.398.096.7103.3103.099.13.2150Ⅰ103.595.793.7100.2102.897.798.93.999.20.7Ⅱ98.693.7103.395.6102.998.498.73.9Ⅲ97.9104.6101.5100.393.4102.1100.03.9臘肉15Ⅰ99.386.7101.399.391.393.395.26.096.20.9Ⅱ89.391.3100.796.7102.798.796.65.4Ⅲ101.396.096.0102.786.798.796.95.930Ⅰ98.396.0102.0100.3102.7101.7100.22.599.60.5Ⅱ96.799.0100.0102.099.398.799.31.8Ⅲ98.3100.7100.794.399.0102.799.32.9150Ⅰ97.9100.795.3102.494.499.998.43.299.71.9Ⅱ98.5105.599.7105.798.3104.0101.93.4Ⅲ95.794.9105.198.3102.097.098.84.0火腿腸15Ⅰ100.092.796.7102.093.387.395.35.693.81.4Ⅱ100.788.090.386.796.797.393.36.1Ⅲ96.093.395.390.789.392.792.92.830Ⅰ100.795.3103.394.0100.7100.799.13.797.81.7Ⅱ97.099.095.3102.395.0101.798.43.2Ⅲ101.796.793.396.393.394.395.93.3150Ⅰ94.398.1105.3103.5100.0102.7100.74.0101.91.6Ⅱ106.1105.1104.995.9106.3104.1103.73.8Ⅲ97.7102.798.7104.599.3105.7101.43.3燒雞15Ⅰ102.789.390.7103.3103.3100.798.36.796.02.6Ⅱ102.0101.398.788.0101.386.796.37.3Ⅲ99.392.094.788.790.794.793.34.030Ⅰ93.394.396.0100.795.0103.397.14.197.70.6Ⅱ96.097.3100.399.097.097.797.91.6Ⅲ96.793.799.099.3100.0100.798.22.7150Ⅰ95.095.697.1104.1102.298.098.73.799.61.1Ⅱ104.599.194.5103.197.7105.9100.84.4Ⅲ95.2105.1101.7104.194.795.099.34.9表13山東院腺苷酸回收率測定結(jié)果待測組織添加濃度(mg/kg)批次回收率（%）批內(nèi)平均回收率（%）批內(nèi)RSD（%）批間平均回收率（%）批間RSD（%）123456豬肉脯15Ⅰ101.396.098.7101.396.795.398.22.794.63.3Ⅱ94.093.390.097.389.390.092.33.4Ⅲ96.7100.086.794.787.394.793.35.730Ⅰ101.395.397.3100.095.394.097.23.097.82.0Ⅱ95.096.793.398.093.3100.796.23.0Ⅲ97.3103.0102.0102.399.096.3100.02.8150Ⅰ100.095.1105.995.3102.6105.0100.64.799.80.9Ⅱ98.395.095.3104.9101.398.598.93.8Ⅲ96.596.3101.298.1105.3102.199.93.6醬牛肉15Ⅰ100.797.3103.393.397.398.798.43.495.32.9Ⅱ92.794.096.794.794.091.393.91.9Ⅲ91.387.392.796.7100.792.093.45.030Ⅰ95.3103.396.3101.398.096.398.43.297.21.3Ⅱ95.393.7101.393.398.793.395.93.5Ⅲ98.393.798.0100.794.099.397.32.9150Ⅰ94.0101.297.4104.494.196.397.94.297.71.4Ⅱ103.395.394.395.2101.1104.899.04.7Ⅲ94.193.8100.096.596.296.796.22.3臘肉15Ⅰ90.797.3100.791.387.398.794.35.693.11.3Ⅱ88.094.793.394.093.394.092.92.6Ⅲ95.389.386.794.792.793.392.03.630Ⅰ101.3101.3103.398.094.098.099.33.498.80.5Ⅱ98.797.397.796.799.3100.098.31.3Ⅲ102.399.794.397.796.3102.098.73.2150Ⅰ94.9103.2105.496.197.7102.199.94.399.50.8Ⅱ98.3103.997.094.7100.797.298.63.3Ⅲ97.099.198.797.4105.3102.9100.13.3火腿腸15Ⅰ