中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2013年
3期
274-277
,共4页
色谱法,液相%质谱法%固相萃取%河豚毒素%海味
色譜法,液相%質譜法%固相萃取%河豚毒素%海味
색보법,액상%질보법%고상췌취%하돈독소%해미
Liquid chromatography%Mass spectrometry%Solid phase extraction%Tetrodotoxin%Seafood
目的 建立石墨化炭黑固相萃取-亲水液相色谱-三重四极杆质谱法测定海产品中河豚毒素的检测方法.方法 将河豚鱼肉、肝和织纹螺肉样品经0.2％乙酸水溶液提取后(肝脏提取液需经HLB固相萃取柱脱脂),再经石墨化炭黑固相萃取柱净化,以含有2.0 mmol/L甲酸铵-0.1％(体积分数)甲酸的95％(体积分数)乙腈水液及含2.0 mmol/L甲酸铵的0.1％甲酸水溶液作为流动相进行梯度洗脱,在色谱柱上(Acquity UPLC BEH Amide,100 mm ×2.1 mm× l.7 μm)实现分离,正离子电喷雾多反应监测方式检测,基质标准外标法定量.结果 河豚鱼肉、肝和织纹螺肉中TTX的线性范围分别为30 ～ 10 000、50 ～ 10 000和30 ～ 10 000 μg/kg,相关系数为0.9963 ～0.9990;检出限分别为10、20和10 μg/kg,定量限分别为30、50和30 μg/kg；平均加标回收率分别为81.5％～93.1％、82.3％～106.0％和83.5％ ～95.2％,RSD分别为2.3％ ～ 11.0％、4.3％ ～14.0％和3.5％～13.0％(n=6).结论 本法简单、准确、灵敏,可应用于河豚鱼和织纹螺中TTX的测定.
目的 建立石墨化炭黑固相萃取-親水液相色譜-三重四極桿質譜法測定海產品中河豚毒素的檢測方法.方法 將河豚魚肉、肝和織紋螺肉樣品經0.2％乙痠水溶液提取後(肝髒提取液需經HLB固相萃取柱脫脂),再經石墨化炭黑固相萃取柱淨化,以含有2.0 mmol/L甲痠銨-0.1％(體積分數)甲痠的95％(體積分數)乙腈水液及含2.0 mmol/L甲痠銨的0.1％甲痠水溶液作為流動相進行梯度洗脫,在色譜柱上(Acquity UPLC BEH Amide,100 mm ×2.1 mm× l.7 μm)實現分離,正離子電噴霧多反應鑑測方式檢測,基質標準外標法定量.結果 河豚魚肉、肝和織紋螺肉中TTX的線性範圍分彆為30 ～ 10 000、50 ～ 10 000和30 ～ 10 000 μg/kg,相關繫數為0.9963 ～0.9990;檢齣限分彆為10、20和10 μg/kg,定量限分彆為30、50和30 μg/kg；平均加標迴收率分彆為81.5％～93.1％、82.3％～106.0％和83.5％ ～95.2％,RSD分彆為2.3％ ～ 11.0％、4.3％ ～14.0％和3.5％～13.0％(n=6).結論 本法簡單、準確、靈敏,可應用于河豚魚和織紋螺中TTX的測定.
목적 건립석묵화탄흑고상췌취-친수액상색보-삼중사겁간질보법측정해산품중하돈독소적검측방법.방법 장하돈어육、간화직문라육양품경0.2％을산수용액제취후(간장제취액수경HLB고상췌취주탈지),재경석묵화탄흑고상췌취주정화,이함유2.0 mmol/L갑산안-0.1％(체적분수)갑산적95％(체적분수)을정수액급함2.0 mmol/L갑산안적0.1％갑산수용액작위류동상진행제도세탈,재색보주상(Acquity UPLC BEH Amide,100 mm ×2.1 mm× l.7 μm)실현분리,정리자전분무다반응감측방식검측,기질표준외표법정량.결과 하돈어육、간화직문라육중TTX적선성범위분별위30 ～ 10 000、50 ～ 10 000화30 ～ 10 000 μg/kg,상관계수위0.9963 ～0.9990;검출한분별위10、20화10 μg/kg,정량한분별위30、50화30 μg/kg；평균가표회수솔분별위81.5％～93.1％、82.3％～106.0％화83.5％ ～95.2％,RSD분별위2.3％ ～ 11.0％、4.3％ ～14.0％화3.5％～13.0％(n=6).결론 본법간단、준학、령민,가응용우하돈어화직문라중TTX적측정.
Objective To develop a rapid hilic ultra performance liquid chromatography (UPLC)-mass spectrum(MS)/MS method for determination of tetrodotoxin in seafood.Methods The sample of muscle and liver of puffer fish and nassarius were extracted with aqueous solution containing 0.2％ (V/V) acetic acid (the extract of liver must be purified through HLB cartridge),and then cleanup of extract was accomplished by solid-phase extraction with a graphitized carbon black cartridge.The analysis of tetrodotoxin was carried out on a chromatographic column (Acquity UPLC BEH Amide,100 mm ×2.1 mm× 1.7 μm) with gradient elution of 95％ (V/V) acetonitrile-H20 both containing 0.1％ (V/V) formic acid and 2.0 mmoL/L ammonium formate,and detected by positive electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode,and quantified by matrix-match standard solution.Results The calibration curves were linear in the range of 30-10 000,50-10 000 and 30-10 000 μg/kg of tetradotoxin in muscle and liver of puffer fish and in muscle of nassarius,respectively.The correlation coefficients were within 0.9963-0.9990.The limits of detection were 10,20 and 10 μg/kg,and that of quantitation were 30,50 and 30 μg/kg for muscle and liver of puffer fish and muscle of nssarius,respectively.The average recoveries were 81.5％-93.1％,82.3％ -106.0％ and 83.5％-95.2％ for tetrodotoxin spiked in muscle and liver of puffer fish and in muscle of nssarius,respectively,with relative standard deviation(RSD) of 2.3％-11％,4.3％-14.0％ and 3.5％-13.0％ (n =6).Conclusion The method was simple,accurate and sensitive,and could be successfully applied to the measurement of tetrodotoxin in puffer fish and nassarius.
