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Optimisation of a high-throughput fluorescamine assay for detection of N-acyl-L-homoserine lactone acylase activity
摘要： N-acyl-L-homoserine lactone (AHL) acylases are a well-known group of enzymes that disrupt quorum sensing in Gram-negative bacteria by degrading AHL signalling molecules. This degradation of signalling molecules (termed 'quorum quenching') has potential uses in the prevention or reduction of biofilm formation and/or bacterial infections. Therefore, there is a great deal of interest in the identification and characterisation of quorum quenching enzymes. Here, we present an optimised fluorescamine-based assay for the detection of AHL acylase activity and demonstrate it can be used in a high-throughput screening format.
关键词： Fluorescamine,Quorum quenching,Fluorescence,High-throughput,AHL acylase
更新于2025-09-23 15:21:01
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Rapid detection of sulfamethoxazole in plasma and food samples with in-syringe membrane SPE coupled with solid-phase fluorescence spectrometry
摘要： In this work, in-syringe membrane solid-phase extraction (MSPE) device was fabricated for the on-site sampling of sulfamethoxazole (SMX) in food samples followed by solid-phase fluorescence spectra analysis. The samples and fluorescamine (FA) were added to a syringe for derivation. Then, the derivative of SMX was extracted by a membrane in the syringe SPE device. Subsequently, the derivative on the membrane was measured immediately without additional elution procedure. The method was successfully applied in plasma, milk, and egg samples for the trace SMX detection, with the recovery of 98 % to 102 %, RSDs from 1 % to 6 %. Compared with liquid chromatography, direct detection of the concentrated analyte significantly improved the sensitivity. Moreover, fluorescamine made it unnecessary to separate SMX from the interference. Consequently, it was a time-saving, low-cost, and easy-operation method, which demonstrated the potential of in-syringe SPE as a promising candidate for on-site analysis.
关键词： Sulfamethoxazole,Food samples,In-syringe solid-phase extraction,Solid-phase fluorescence,Fluorescamine
更新于2025-09-23 15:19:57
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The convenient use of fluorescamine for spectrofluorimetric determination of midodrine hydrochloride in pure form and its tablets formulation: Application to content uniformity testing
摘要： A novel sensitive and simple spectrofluorimetric method was developed then validated for determination of midodrine in both its authentic pure form and its tablets. This method is based on the reaction between midodrine's aliphatic primary amine moiety with fluorescamine reagent, using borate buffer at pH 7.8 and yielding a highly fluorescent product whose fluorescence intensity was measured at 462 nm after excitation at 388 nm. This method represents the first attempt for determination of midodrine spectrofluorimetrically. A calibration curve was constructed showing that the linear range was 0.2–3.0 μg/ml. The limit of detection and limit of quantitation values were 0.06 and 0.19 μg/ml respectively. The correlation coefficient (r) and the determination coefficient (r2) values were 0.9992 and 0.9984 respectively. The proposed method was validated according to ICH guidelines and successfully applied for determination of midodrine in its tablets with an overall % recovery of 99.56 ± 0.95. Finally, the presented method was adapted to study the content uniformity test according to United States Pharmacopeia guidelines.
关键词： content uniformity testing,fluorescamine,midodrine hydrochloride,spectrofluorimetric determination
更新于2025-09-19 17:15:36