鱼露中章鱼胺含量分析及衍生化产物结构推断

Determination of octopamine in fish sauce and deduction structure of corresponding derived products

利用衍生化法建立了准确的鱼露中章鱼胺分析方法,同时结合高分辨质谱对其衍生产物准确定性。鱼露试样以1,7-二氨基庚烷为内标,采用丹磺酰氯衍生法使章鱼胺及其内标在碱性条件下与丹磺酰氯生成衍生物,乙醚提取,产物经C_(18)色谱柱分离,高效液相色谱-紫外检测器检测,内标法定量。章鱼胺在线性1~20mg/L范围内线性关系良好,检出限为0.3mg/L,定量限为1.0mg/L,回收率介于70.6~104.7%之间,精密度小于9%。在此基础上运用超高效液相色谱-二极管阵列检测器-四极杆飞行时间串联质谱仪(UHPLC-PDA-Q-TOF-MS)对产物结构进行推断,进一步明确产物的色谱峰峰位及衍生化产物形成机理,并将所开发的方法应用于市售鱼露样品中章鱼胺的定量定性分析。该方法定性定量准确,可避免假阳性事件的发生。

An accurate analytical method for octopamine in fish sauce was established by derivation method,and high-resolution mass spectrometry was used to qualitative analysis its derivative products.1,7-diaminoheptane was used as the internal standard.Firstly,the octopamine and internal standard were derivatized with dansulfoyl chloride under alkaline conditions,and the corresponding derived products were extracted with diethyl ether.Then the products were separated by C_(18) chromatography column,detected by high-performance liquid chromatography with ultraviolet detector,and quantified by internal standard method.Octopamine had a good linear relationship with the correlation coefficients(R^(2)) higher than 0.99 in the linear range of 1-20mg/L.The limit of detection was 0.3mg/L and limit of quantification was 1.0mg/L.The recoveries of octopamine were in the range of 70.2%-113% with the relative standard deviations(RSDs) was less than 9%.On this basis,the structure of the derived product was inferred using ultra-high performance liquid chromatography coupled to diode array detector quadrupole time of flight mass spectrometry(UHPLC-PDA-Q-TOF-MS).The rentention time and formation mechanism of derived product were further clarified.The derived products of octopamine and 1,7-diaminoheptane were formed through reacting with dansulfoyl chloride in a ratio of 1:2,respectively.The method can be used for quantitative and qualitative analysis of octopamine in commercially available fish sauce samples,avoiding the occurrence of false positive events.