用铁(Ⅱ)-1,10-邻菲啰啉-大黄素体系的共振瑞利散射光谱法测定生物样品中大黄素  

Determination of emodin by resonance Rayleigh scattering spectral method* of Fe(Ⅱ)-1,10-phen-emodin system in biological sample

作　　者：王二女 陈科平 杨季冬[1,3] 

机构地区：[1]西南大学化学化工学院,重庆400715 [2]重庆市涪陵环境监测中心,重庆408000 [3]三峡学院化学与环境科学学院,重庆404000

出　　处：《药物分析杂志》2015年第6期1010-1015,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(No.21175015;No.21475014)资助

摘　　要：目的：建立共振瑞利散射法测定大黄素的含量。方法：在p H 7.3的B-R缓冲溶液中,铁（Ⅱ）与1,10-邻菲啰啉形成稳定的1∶3的配合物,再与大黄素（EMO）结合形成1∶2的离子缔合物。铁（Ⅱ）-1,10-邻菲啰啉溶液：称取0.278 0 g Fe SO4·7H2O溶于70 m L水中,加入邻菲啰啉（1,10-phen）0.595 0 g,加入适量的抗坏血酸,稀释至100 m L,得到1×10^-2mol·L^-1的储备液,用时稀释成2.0×10^-3mol·L^-1的工作液。室温下,于10 m L比色管中依次加入p H 7.3的B-R缓冲溶液2 m L,适量的EMO标准溶液,以及2.0×10^-3mol·L^-1Fe（phen）2＋3溶液1.2 m L。每加一种试剂后混合均匀,用二次蒸馏水定容至刻度,摇匀,静置10 min后,在荧光分光光度计上以λex=λem方式进行同步扫描,记录共振瑞利散射（RRS）光谱,以λem=2λex和λem=1/2λex进行扫描,分别测量不同入射波长（λex）下的散射强度ISOS和IFDS,然后分别以ISOS和IFDS对应的波长作图,得到二级散射（SOS）光谱和倍频散射（FDS）光谱。分别在各自的最大散射波长处测量样品和试剂空白的散射强度IRRS,ISOS,IFDS及I0RRS,I0SOS,I0FDS,ΔI=I-I0。结果：体系的RRS、SOS和FDS显著增强并出现新的散射峰,相应的最大散射峰分别位于349、684和351 nm。EMO的质量浓度在0.8-10.4μg·m L-1时,与RRS、SOS和FDS的散射强度呈良好的线性关系,其检出限（3σ）依次分别为10.1、32.8、28.6 ng·m L^-1。血样和尿样（各3批）中测定EMO的回收率分别在94.9%-102.4%和99.4%-102.7%之间,RSD分别在1.5%-3.1%和1.1%-3.0%之间。将本法与紫外可见分光光度法比较,结果满意。结论：经方法学验证,体系的共振瑞利散射方法可用于尿样和血清中大黄素的测定。Objective：To determine the content of emodin by resonance Rayleigh scattering method.Methods：In p H 7.3 Britton-Robinson（BR） buffer medium,the interaction between Fe2 ＋and 1,10-phenanthroline（1∶ 3） occurred rapidly,which was further merged with emodin（EMO） into a 1∶ 2 ion-association complex.A 1 × 10-2mol·L-1stock solution of Fe（phen）2 ＋3was prepared by dissolving 0.278 0 g of Fe SO4·7H2O in 70 m L of doubly distilled water,to which 0.595 0 g of phenanthroline and appropriate amounts of ascorbic acid were added,and then diluted to the mark in a 100 m L brown volumetric flask.The working solution of 2.0 × 10-3mol·L-1Fe（phen）2 ＋3was obtained by diluting the stock solution with water.At room temperature,a 10 m L calibrated flask was added with2.0 m L of p H 7.3 BR buffer solution,suitable amounts of EMO and 1.2m L of 2.0 × 10-3mol·L-1Fe（phen）2 ＋3solution.The mixture was diluted to the volume mark with water and mixed thoroughly.After waiting for 10 min,the resonance Rayleigh scattering（RRS） spectra of the system was recorded with synchronous scanning at λem=λex,and the second order scattering（SOS） and frequency doubling scattering（FDS） spectra were recorded by scanning at λem= 2 λexand λem= 1/2 λex,respectively.Then,the scattering intensity IRRS,ISOSand IFDSfor the reaction product and I0 RRS,I0SOSand I0 FDSfor the reagent blank at their maximum wavelengths were measured,ΔIRRS=IRRS-I0 RRS,ΔISOS= ISOS-I0 SOSand ΔIFDS =IFDS-I0 FDS.Results：The intensities of RRS,SOS and FDS were enhanced greatly and the new RRS,SOS and FDS spectra appeared.Their maximum wavelengths were located at 349 nm,684 nm and 351 nm,respectively.The linear ranges were all 0.8-10.4 μg·m L-1for RRS,SOS and FDS,and the detection limits were 10.1 ng·m L-1,32.8 ng·m L-1and 28.6 ng·m L-1,separately.The recovery of serum and urine samples（3 batches for each） ranged from 94.9 % to 102.4 % and 99.4 % to 102.7 %,RSD from1.5 % to 3.1 % and 1.1 % to 3.0 %,respectively.Compared wit

关 键 词：大黄素 铁(Ⅱ)-1 10-邻菲啰啉 离子缔合物 共振瑞利散射(RRS) 二级散射(SOS) 倍频散射(FDS) 生物样品测定 

分 类 号：R917[医药卫生—药物分析学]