2.5. Real Sample Detection of Gentamicin
This protocol is extracted from research article:
A Paper-Based Colorimetric Aptasensor for the Detection of Gentamicin
Biosensors (Basel), Jan 21, 2021; DOI: 10.3390/bios11020029

To explore the practical applicability of the colorimetric and paper sensor, skimmed milk (2%) spiked with gentamicin was used. Milk, being a complex matrix of proteins and caseins was first pretreated to remove these interfering materials described in previous studies [26]. In a typical experiment, 1 mL of milk sample was spiked with various concentrations on gentamicin before the pretreatment process. To this, 1 M HCl (pH 4.5) was added to precipitate the caseins, followed by centrifugation at 12,000 rpm for 5 min. The supernatant was separated and transferred to a fresh centrifuge tube, to which 300 µL methanol was added and the centrifugation process was repeated. The final clear supernatant was allowed to interact with ssDNA modified gold nanoparticles in the ratio 1:10. Spectral sweep data were obtained from 200–700 nm and the peak absorbance ratio of A640/A520 was calculated.

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