Cao Jie, Li Mingjie, Chen Xiao-Ying
Scientific Research and Experiment Center, Fujian Police College Fuzhou 350007 China
Fuzhou University Postdoctoral Research Station of Chemistry, Fuzhou University Fuzhou 350108 China.
RSC Adv. 2022 Oct 26;12(47):30529-30538. doi: 10.1039/d2ra05202d. eCollection 2022 Oct 24.
In this paper, a sensitive and specific competitive fluorescence immunoassay (CFIA) method was developed for the qualitative and quantitative analysis of ketamine (KET). A novel competitive model in which ketamine hapten (KET-BSA), coated on microporous plates, competed with ketamine antigen (KET-Ag) in actual samples to bind fluorescein isothiocyanate-labeled antibody (KET-Ab) could be used for rapid and indirect quantitative analysis of KET in human urine, blood, or sewage. In the CFIA method, KET concentration in the sample negatively correlated with the detected fluorescence intensity. The linear correlation coefficient of the competitive quantitative equation was 0.992, the linear range was 0.01-0.5 μg mL, and the limit of detection (LOD) was 0.1 pg mL. The specificity results showed that the cross-reaction rate of norketamine was less than 10%. Recoveries of spiked samples at low, medium, and high concentrations ranged from 96% to 117%. The CFIA method and classical gas chromatography-tandem mass spectrometry (GC-MS/MS) were used to detect the actual samples simultaneously. The relative deviation of the quantitative results was less than 10%. The LOD value of KET by CFIA was four orders of magnitude lower than that by GC-MS/MS. Additionally, CFIA had great advantages over GC-MS/MS in terms of sample pretreatment and economic investment. In conclusion, this study provided a targeting detection platform for KET, which achieved a rapid, portable, and sensitive analysis of trace KET in various materials.
本文建立了一种灵敏、特异的氯胺酮（KET）竞争性荧光免疫分析（CFIA）方法，用于KET的定性和定量分析。一种新型的竞争模型，即包被在微孔板上的氯胺酮半抗原（KET-BSA）与实际样品中的氯胺酮抗原（KET-Ag）竞争结合异硫氰酸荧光素标记的抗体（KET-Ab），可用于快速、间接定量分析人尿液、血液或污水中的KET。在CFIA方法中，样品中KET浓度与检测到的荧光强度呈负相关。竞争定量方程的线性相关系数为0.992，线性范围为0.01 - 0.5 μg/mL，检测限（LOD）为0.1 pg/mL。特异性结果表明，去甲氯胺酮的交叉反应率小于10%。低、中、高浓度加标样品的回收率在96%至117%之间。同时采用CFIA方法和经典的气相色谱 - 串联质谱（GC-MS/MS）检测实际样品。定量结果的相对偏差小于10%。CFIA法检测KET的LOD值比GC-MS/MS法低四个数量级。此外，CFIA在样品预处理和经济投入方面比GC-MS/MS具有很大优势。总之，本研究为KET提供了一个靶向检测平台，实现了对各种材料中痕量KET的快速、便携和灵敏分析。
