Detection of Ketamine and Norketamine Using an Aptamer-Functionalized Gra-phene Oxide Fluorescent Sensor
10.12116/j.issn.1004-5619.2025.350409
- VernacularTitle:基于核酸适配体功能化氧化石墨烯荧光传感器检测氯胺酮及去甲氯胺酮
- Author:
Li-Xia WEI
1
;
Bo LIU
;
Xiao-Yuan YANG
;
Xi ZHANG
;
Yi-Feng LAN
;
Chao ZHANG
;
Juan JIA
;
Dan ZHANG
;
Zhi-Wen WEI
;
Ke-Ming YUN
;
Zhe CHEN
Author Information
1. 山西医科大学法医学院,山西 晋中 030600;法庭毒物分析公安部重点实验室,山西 晋中 030600;法医学山西省重点实验室,山西 晋中 030600
- Keywords:
forensic medicine;
toxicological analysis;
ketamine;
norketamine;
graphene oxide-systematic evolution of ligand by exponential enrichment(GO-SELEX);
aptamer;
chemical conjugation;
fluores-cent sensor
- From:
Journal of Forensic Medicine
2025;41(4):326-339
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an aptamer-functionalized carboxylated graphene oxide(CGO)fluo-rescent sensor to achieve highly sensitive and specific detection of ketamine(KET)and its metabolite norketamine(NK)using an aptamer capable of simultaneously recognizing KET and NK.Methods A specific aptamer for simultaneous recognition of KET and NK was screened using graphene oxide-sys-tematic evolution of ligand by exponential enrichment(GO-SELEX)and molecular docking tech-niques.The aptamer,labeled with Cy5 fluorescence,was chemically conjugated to CGO to construct an aptamer-functionalized CGO fluorescent sensor.By optimizing detection conditions,including the mass concentration of CGO,aptamer concentration,reaction temperature,and incubation time,quantita-tive analysis of the target analytes was achieved using the ratio of fluorescence intensity changes be-fore and after target addition.The stability of the sensor in biological matrices was evaluated by moni-toring fluorescence intensity changes over incubation time in blank blood and urine,in comparison with the traditional physical adsorption-based CGO fluorescent sensor.Spiked recovery experiments in blank blood and urine were conducted to compare performance with that of HPLC-MS/MS.Results A specific aptamer A5 was selected and chemically conjugated with CGO to construct the aptamer-functionalized CGO fluorescent sensor.Under optimized conditions,the proposed fluorescent sensor ex-hibited a linear detection range of 1.0-5.0 ng/mL for KET,with a limit of detection(LOD)of 0.86 ng/mL;while for NK,the linear detection range was 1.0-5.0 ng/mL,with an LOD of 0.70 ng/mL.Com-pared with the CGO fluorescent sensor constructed via physical adsorption,this sensor demonstrated greater stability in blood and urine.The spiked recovery rates of KET and NK in blank blood and urine ranged from 81.50%to 110.03%,exhibiting detection performance comparable to that of HPLC-MS/MS.Conclusion The aptamer screening method offers a novel approach for selecting aptamers tar-geting drugs and their metabolites.The constructed aptamer-functionalized CGO fluorescent sensor pro-vides an efficient and reliable strategy for the high-performance detection of KET and NK.
