CAJ | 학술논문

在稀碱条件下,河鲀毒素(TTX)与次溴酸钠和尿素于一定温度下反应生成具有荧光性能的物质,该反应与已报道的TTX在强碱下生成C-9碱的碱解不同。本研究拟对该衍生反应所需条件进行详细研究,并尝试依据该反应建立鲀一种新型的河毒素荧光检测方法。荧光扫描光谱显示,产物最大激发波长(EX)为233 nm,最大发射波长(EM)为370 nm。该反应对pH要求比较严格,反应最适pH在11.6~11.9之间。对样品进行衍生检测获得最强荧光信号的条件为：次溴酸钠0.036~0.09 mol/L、尿素10~50 g/L、碳酸钠0.2~0.3 mol/L,温度75℃。样品检测结果显示,此衍生反应具有高度专一性,信号强度与 TTX 含量成正比,在0.01~10μg/mL 质量浓度范围内线性方程为y=109.17x+0.3965,线性相关系数为0.999。依据该衍生反应建立柱前衍生–高效液相色谱荧光检测法检测水产品中河鲀毒素含量,检测限达到20μg/kg,平行样品检测相对标准偏差为0.44%~7.25%,准确度高。对河鲀、虾虎鱼和织纹螺产品检测表明,该反应特异性强,不易出现假阳性,鲀可用于河毒素检测的确证。
재희감조건하,하돈독소(TTX)여차추산납화뇨소우일정온도하반응생성구유형광성능적물질,해반응여이보도적TTX재강감하생성C-9감적감해불동。본연구의대해연생반응소수조건진행상세연구,병상시의거해반응건립돈일충신형적하독소형광검측방법。형광소묘광보현시,산물최대격발파장(EX)위233 nm,최대발사파장(EM)위370 nm。해반응대pH요구비교엄격,반응최괄pH재11.6~11.9지간。대양품진행연생검측획득최강형광신호적조건위：차추산납0.036~0.09 mol/L、뇨소10~50 g/L、탄산납0.2~0.3 mol/L,온도75℃。양품검측결과현시,차연생반응구유고도전일성,신호강도여 TTX 함량성정비,재0.01~10μg/mL 질량농도범위내선성방정위y=109.17x+0.3965,선성상관계수위0.999。의거해연생반응건립주전연생–고효액상색보형광검측법검측수산품중하돈독소함량,검측한체도20μg/kg,평행양품검측상대표준편차위0.44%~7.25%,준학도고。대하돈、하호어화직문라산품검측표명,해반응특이성강,불역출현가양성,돈가용우하독소검측적학증。
Tetrodotoxin (TTX), one of the most toxic natural neurotoxins found in several kinds of fishery products, can cause TTX poisoning. Therefore, detection methods for routine analysis of TTX should be developed. We found that TTX could be converted into a new compound with fluorescence properties by reaction with sodium bromate (NaBrO) and urea. The reaction was sensitive to the acidity level of the solution and the pH value needed to be strictly controlled between 11.6 and 11.9. The fluorescence scanning spectrum showed the fluorescent compound had maximum excitation wavelength of 233 nm and emission wavelength of 370 nm, and was not the product of alkaline degradation (quina-zoline). Hence, a new type of fluorescence detection method was developed for identifying TTX in aquatic products. After reaction with NaBrO (0.036–0.09 mol/L) and urea (10–50 g/L) in 0.2–0.3 mol/L sodium carbonate solution (Na2CO3) at 75℃, the TTX derivative solution was neutralized with 15% phosphoric acid and then detected by re-verse-phase high-performance liquid chromatography with a fluorescence detector. The result was quantified by the standard addition method. The detection limit was 20μg/kg and the relative standard deviation was 0.44%–7.25%. The method was specific, accurate, and sensitive, and can be used for identification of TTX determination.