固相萃取-柱前衍生-高效液相色谱法测定水体中9种生物胺的含量  

Determination of 9 Biogenic Amines in Water Body by High Performance Liquid Chromatography Combined with Solid Phase Extraction and Pre-Column Derivatization

作　　者：姚燕红 吴平谷[1] 王军淋[1] 冯靓[1] 赵永信[1] 张念华[1] YAO Yanhong;WU Pinggu;WANG Junlin;FENG Liang;ZHAO Yongxin;ZHANG Nianhua(Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou 310051,China)

机构地区：[1]浙江省疾病预防控制中心,杭州310051

出　　处：《理化检验(化学分册)》2025年第4期400-406,共7页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

摘　　要：提出了固相萃取-柱前衍生-高效液相色谱法测定水体中9种生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、章鱼胺、酪胺、亚精胺和精胺)含量的方法。取水样10 mL置于15 mL离心管中,加入100 mg·L^(-1)混合内标使用液250μL和50 mmol·L^(-1)盐酸溶液3 mL,混匀后以流量3 mL·min^(-1)通过固相萃取柱(弱阳离子交换柱),依次加入2 mL水、2 mL甲醇、4 mL含2 g·L^(-1)氢氧化钠的甲醇溶液,收集洗脱液置于15 mL离心管(预先加入1 mol·L^(-1)盐酸溶液250μL)中,氮吹至近干,加入300μL饱和碳酸氢钠溶液,加水至1 mL,再加入1 mL缓冲溶液(p H 10.5),混匀。加入10 g·L^(-1)丹磺酰氯衍生溶液1 mL,涡旋混匀,于60℃加热15 min,冷却至25℃,加入200μL氨水,于60℃加热10 min,冷却至25℃,加入0.5 g氯化钠和0.5 mL乙腈,混匀,离心1 min,取上清液过0.22μm滤膜,测定滤液中9种生物胺的含量。以Waters XBridge C18色谱柱为固定相,以不同体积比的含0.1%(体积分数)乙酸的0.01 mol·L^(-1)乙酸铵溶液-乙腈混合溶液为流动相进行梯度洗脱,内标法定量。结果表明,9种生物胺的质量浓度在1.0~100.0 mg·L^(-1)内与对应的衍生物峰面积与内标峰面积比值呈线性关系,检出限(3.143s)为0.03~0.08 mg·L^(-1)。按照标准加入法进行回收试验,回收率为76.4%~107%,测定值的相对标准偏差(n=6)均不大于5.0%。方法用于18份实际水样的分析,普遍检出苯乙胺,检出量为0.03~0.13 mg·L^(-1)。A method for determination of 9 biogenic amines including tryptamine,phenylethylamine,putrescine,cadaverine,histamine,octopamine,tyramine,spermidine and spermine in water body by high performance liquid chromatography combined with solid phase extraction and pre-column derivatization was proposed.10 mL of water sample was taken and placed in a 15 mL-centrifuge tube.250μL of 100 mg·L^(-1) mixed internal standard solution and 3 mL of 50 mmol·L^(-1) hydrochloric acid solution were added.The mixture was mixed and passed through a solid-phase extraction column(weak cation exchange column)at a flow rate of 3 mL·min^(-1).2 mL of water,2 mL of methanol and 4 mL of methanol solution containing 2 g·L^(-1) sodium hydroxide were added in sequence.The eluent was collected and placed in a 15 mL-centrifuge tube(pre-added with 250μL of 1 mol·L^(-1) hydrochloric acid solution),and blown to near drynesss by nitrogen.300μL of saturated sodium bicarbonate solution was added,and the solution was made its volume up to 1 mL with water.1 mL of buffer solution(pH 10.5)was added and the mixture was mixed well.1 mL of 10 g·L^(-1) dansyl chloride derivative solution was added,and the solution was vortexed and heated at 60℃for 15 min.The solution was cooled to 25℃,and 200μL of ammonia was added.The solution was heated at 60℃for 10 min and cooled to 25℃.0.5 g of sodium chloride and 0.5 mL of acetonitrile were added.The mixture was mixed well and centrifuged for 1 min.The supernatant was passed through a 0.22μm filter membrane,and 9 biogenic amines in the filtrate was determined using Waters XBridge C18 chromatographic column as the stationary phase and the mixed solution composed of 0.01 mol·L^(-1) ammonium acetate solution containing 0.1%(volume fraction)acetic acid and acetonitrile at different volume ratios as mobile phase for gradient elution,with the internal standard method for quantification.As shown by the results,linear relationships between the ratios of the corresponding derivative peak area to the internal stan

关 键 词：固相萃取 柱前衍生 高效液相色谱法 水体 生物胺 

分 类 号：O657.7[理学—分析化学]