上转发光技术快速检测尿液中氯胺酮

Rapid Detection of Ketamine in Urine Based on Up-converting Phosphor Technology

目的利用上转发光免疫层析技术（UPT-LF）建立一种可在现场快速定量检测尿液中氯胺酮（Ketamine,KET）的方法并对其进行系统评价。方法利用竞争法免疫层析原理,以上转发光材料（UCP）作为生物示踪物,建立快速检测尿液中KET含量的上转发光免疫层析定量检测方法（KET-UPT-LF）,并通过专用的便携UPT生物传感器分析进而计算出氯胺酮浓度。通过系列浓度标准品检测评价其敏感性、精密性及线性响应。收集经LC-MS鉴定的现场尿液样本,同时进行KET-UPT-LF及胶体金的检测,一方面通过与胶体金定性检测结果的比较来评价KET-UPT-LF对于实际尿液样本中KET的定性检测能力,另一方面通过与LC-MS的定量检测结果比较来评价KET-UPT-LF定量检测性能。结果通过对系列浓度标准品检测结果分析,KET-UPT-LF敏感性为93.75 ng/m L,线性范围为93.75~6 000 ng/m L（r=-0.99448,P〈0.0005）。在现场样品检测评价中,同一样本的KET-UPT-LF和胶体金结果在定性分析,其结果是一致的。KET-UPT-LF与定量确证方法 LC-MS的定量检测结果经t检验无显著差异。结论本研究建立的氯胺酮上转发光免疫层析定量检测方法（KET-UPT-LF）,在满足胶体金等快筛试剂快速简便的基础上,进一步实现了现场快速定量检测,且用户友好性优于LC-MS等定量确证方法,为现场快速定量检测尿液中的毒品提供了技术保障。

Objective To develop and evaluate an up-converting-phosphor-technology-based lateral flow（UPT-LF） assay for fast quantitative detection of Ketamine（KET） in urine. Methods KET-UPT-LF was established with the up-converting phosphor（UCP） nano-particles as the bio-label of competitive-mode-based LF assay. The results were analyzed by a specific portable UPT-based biosensor with which the concentration of ketamine was measured. The detectivity, precision and linearity were evaluated by testing the samples with serial standard concentrations. The urine samples were used to assess the KETUPT-LF＇s both qualitative and quantitative detection performance with immune colloidal gold technique and LC-MS as the respective reference. Results According to the analysis of urine samples detection, the sensitivity of KET-UPT-LF was 93.75 ng/m L and linearity range was 93.75-6000 ng/m L（r =-0.99448,P〈 0.0005）. The performance of qualitative and quantitative detection could meet the requirements of the on-site rapid detection of KET in urine. There was no significant difference between KET-UPT-LF and immune colloidal gold technique for qualitative detection and neither for quantitative detection between KET-UPT-LF and LC-MS. Conclusion The method of KET-UPT-LF for urine test is not only as quick and easy as colloidal gold technique but also provides quantitative detection with a more user-friendly feature compared to LC-MS, offering an alternative to realize on-site rapid detection of KET in urine quantitatively.