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    超高效液相色谱-串联质谱法快速测定蔬菜中4种异噻唑啉酮类化合物的含量

    Rapid Determination of 4 Isothiazolinones in Vegetables by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

    • 摘要: 提出了超高效液相色谱-串联质谱法(UHPLC-MS/MS)快速测定蔬菜中5-氯-2-甲基-4-异噻唑啉-3-酮(CMIT)、2-甲基-4-异噻唑啉-3-酮(MIT)、1,2-苯并异噻唑啉-3-酮(BIT)、2-正辛基-4-异噻唑啉-3-酮(OIT)等4种异噻唑啉酮类化合物含量的方法。取10.0 g已匀浆的蔬菜样品,用10 mL乙腈超声提取30 min,离心,残渣重复提取一次,合并提取液,加入5 g氯化钠,振荡、静置后,于40 ℃氮吹至近干,用2 mL 10%(体积分数,下同)甲醇溶液溶解残渣。所得溶液过已活化的HLB固相萃取小柱,用5 mL 10%甲醇溶液淋洗,6 mL甲醇洗脱。洗脱液于40 ℃氮吹至近干后,用甲醇定容至1 mL,过0.22 μm有机滤膜,滤液在ACQIUTY UPLC BEH SHIELD RP18色谱柱上分离,以不同体积比的甲醇-水混合液为流动相进行梯度洗脱,质谱分析采用多反应监测采集模式,电喷雾正离子扫描模式,基质匹配法绘制工作曲线。结果表明,4种异噻唑啉酮类化合物的质量浓度在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)分别为0.5,0.5,0.5,0.025 μg·kg-1。在不同蔬菜基质中进行3个浓度水平的加标回收试验,回收率为68.8%~81.3%,测定值的相对标准偏差(n=6)均小于9.0%。方法用于20种蔬菜样品分析,仅一组韭菜样品中检出BIT,检出量为2.3 μg·kg-1

       

      Abstract: A method for rapid determination of 4 isothiazolinones in vegetables, including 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT), 2-methyl-4-isothiazolin-3-one (MIT), 1,2-benzoisothiazolin-3-one (BIT), and 2-n-octyl-4-isothiazolin-3-one (OIT), by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was proposed. The homogenized vegetable sample (10.0 g) was taken, extracted by ultrasound in 10 mL of acetonitrile for 30 min, and then centrifuged. The residue was extracted again, which was combined with the extraction solution. 5 g of sodium chloride was added. After oscillation and settlement, the extraction solution was blown to near dryness by nitrogen at 40 ℃. The residue was dissolved in 2 mL of 10% (volume fraction, the same below) methanol solution. The obtained solution were purified by HLB solid phase extraction column, which was eluted with 5 mL of 10% methanol solution and 6 mL of methanol. The eluate was blown to near dryness by nitrogen at 40 ℃, and made its volume up to 1 mL with methanol, which was passed through 0.22 μm organic filter membrane. The filtrate was separated on ACQIUTY UPLC BEH SHIELD RP18 column, with a mixture of methanol and water at different volume ratios for gradient elution. Multiple reaction monitoring mode and electrospray ionization positive ion mode were adopted in MS analysis, and matrix matching method was used for drawing the working curves. As shown by the results, linear relationships between the corresponding peak areas and mass concentrations of 4 isothiazolinones were found in definite ranges, with detection limits (3S/N) of 0.5,0.5,0.5,0.025 μg·kg-1, respectively. Test for recovery was made at 3 concentration levels in different vegetable matrices, giving results in the range of 68.8%-81.3%, with RSDs (n=6) of the determined values less than 9.0%. This method was used for the analysis of 20 vegetable samples, and BIT was detected in only one group of leek samples, with the detected amount of 2.3 μg·kg-1.

       

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