【摘要】 利用柠檬酸-金纳米粒子(citrate-AuNPs)与吲哚环之间的特异性吸附作用,建立了citrate-AuNPs固相萃取-高效液相色谱同时测定人血浆中色氨酸、5-羟色胺、5-羟吲哚乙酸和褪黑素的方法。在优化的萃取和分离条件下,采用C18柱为色谱分离柱,由甲醇(A)和40 mmol·L-1磷酸二氢钠-磷酸(pH 4.0)组成流动相(B),梯度洗脱程序(v/v):0~4 min 20%A;4~15 min 55%A;荧光检测λex=254 nm,λem=338 nm。四种吲哚胺的浓度分别在1~500 nmol·L-1、0.2~400 nmol·L-1、0.4~800 nmol·L-1和0.1~200 nmol·L-1内具有良好线性关系,相关系数均高于0.99。方法检出限(S/N=3)在0.1 nmol·mL-1~0.2 nmol·mL-1之间,回收率在86%~105%之间。该方法能有效降低生物样品的基体干扰,并通过富集提高传统检测方法的灵敏度。将该方法应用于老年痴呆病(AD)高发组和对照组组人血浆测定,结果显示:AD高发组四组分水平均显著低于对照组(P<0.05),与人脑脊液的规律一致。更多还原

【Abstract】 Based on the specific interactions between citrate-gold nanoparticles(citrate-AuNPs) and indole ring,a citrate-AuNPs solid phase extraction with high performance liquid chromatography(HPLC) method for simultaneous determination of 5-hydroxytryptamine(5-HT),5-hydroxy-indoleacetic acid(5-HIAA),tryptophan(Trp) and melatonin(Mel) in human plasma was established.Under the optimal conditions,the separation was obtained on the C18column using a mixture of methanl(A) and 0.04 mol·L-1NaH2PO4-H3PO4(pH 4.0) as mobile phase(B) with gradient elution:0 ~ 4 min 20% A(v/v),4 ~ 15 min 55% A(v/v).The wavelength of fluorescence detector was set at λex = 254 nm and λem = 338 nm.The linear ranges were found between peak area and concentration in the range of 1 ~ 500 nmol·L-1for 5-HT,0.2 ~ 400 nmol·L-1for 5-HIAA,0.4 ~ 800 nmol·L-1for Trp and 0.1 ~200 nmol·L-1for Mel,with linear regression coefficients all above 0.99.The limits of detection(S/N = 3) for the indoleamines were between 0.1 nmol·mL-1and 0.2 nmol·mL-1and the recoveries were between 86% and 105%.Due to significant interference reduction of endogenous substances in plasma and subsequently improved the selectivity and sensitivity of traditional HPLC method,the presented method was applied for the simultaneous detection of indoleamines in plasma samples of high risk of Alzheimer’s desease group and the controls.The results showed that four indoleamines levels of the high risk group were all significant lower than the controls(P<0.05),and agreed with the rule of cerebrospinal fluid.更多还原