A simple, fast, and highly selective and sensitive colorimetric assay to detect nanomolar levels of spermine in human urine (healthy donors, cancer patients) is reported. This assay is based on the absence of a competitive organic capping on the gold nanoparticles together with the high affinity of the amine groups of the analyte for the nanoparticle surface.

A procedure for supporting silver nanoparticles (AgNPs) on nylon is proposed. Besides, the membrane has been developed as a solid-phase colorimetric plasmonic sensor for volatile sulfide compounds (VSCs) like H 2 S, CH 3 SH, and (CH 3 ) 2 S. AgNP behavior in the membrane has been studied by UV−vis diffuse reflectance spectrometry, Raman spectrometry, High-resolution transmission electron microscopy (HR-TEM), and Scanning electron microscopy (SEM). The sensor responded by changing its color from yellow in absence of VSCs to several orange/brown colors in the function of VSC concentration as occurs in solution; an increase in the hydrodynamic diameter, estimated by both asymmetrical flow field-flow fractionation (AF4) coupled on line to Dynamic light scattering (DLS) detector and batch DLS, is achieved when sulfide is added to the citrate-capped AgNPs. Diffuse reflectance spectrometry and processed digital images obtained with a smartphone have been used as measurements and several transformations for quantitation are proposed; a linear concentration range of hydrogen sulfide from 150 to 1000 ppbv and a detection limit (LOD) of 45 ppbv were achieved, measuring after 10 min of the sensor exposition to the hydrogen sulfide atmosphere (2 L) for humidity percentages between 50 and 96% and room temperature. Satisfactory results in terms of precision (<10%) and selectivity were obtained. The new sensor reported was stable, sensitive, inexpensive, disposable, safe, and user-friendly. Furthermore, it has successfully been applied to determine VSCs expressed as hydrogen sulfide in breath samples (2 L and 250 mL) as a proof of concept. The limit of detection can be improved by increasing the exposition time, if necessary.

Sarcosine, a potential biomarker for prostate cancer, can be detected in a solid state enzyme based biosensor using sarcosine oxidase, with particle immobilised reagents. A novel fusion protein of the fluorescent protein, mCherry, sarcosine oxidase (SOx), and the polypeptide R5 (R52-mCherry-SOx-R5-6H), was explored, which allowed self-immobilization on silica microparticles and long-term (90 days +) retention of activity, even at room temperature. In contrast, commercial wildtype SOx lost activity in a few days. A silica-R52-mCherry-SOx-R5-6H microparticle sensor for determination of sarcosine in urine, linked the SOx coproduct, H2O2, to a measurement catalysed by horseradish peroxidase (HRP) immobilised on silica, in the presence of Amplex Ultrared (AR) to generate fluorescence at 582 nm. Silica microparticles carrying all the reagents (R52-mCherry-SOx-R5-6H, HRP and AR) were used to produce a silica-microparticle biosensor which responded to sarcosine at micromolar levels. Interference by amino acids and uric acid was examined and it was found that the silica-reagent carrying system could be calibrated in urine and responded across the clinically relevant concentration range. This contrasted with similar assays using commercial SOx, where interference inhibited the sarcosine signal measurement in urine. The microparticle biosensor was tested in urine from healthy volunteers and prostate cancer patients, showing higher concentrations of sarcosine in cancer patients consistent with previous reports of elevated sarcosine levels.

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