Development of a µPAD for the point-of-care testing of serum glutamic oxaloacetic transaminase (SGOT).

A wax-patterned paper analytical device (µPAD) has been developed for point-of-care colourimetric testing of serum glutamic oxaloacetic transaminase (SGOT). The detection method was based on the transamination reaction of aspartate with α-ketoglutarate, leading to the formation of oxaloacetate which reacts with the reagent Fast Blue BB salt and forms a cavern pink colour. The intensity of the cavern pink colour grows as the concentration of SGOT increases. UV-visible spectroscopy was utilized to optimize reaction conditions, and the optimized reagents were dropped onto the wax-patterned paper. The coloured PADs, after the addition of SGOT, have been photographed, and a colour band has been generated to correlate the SGOT concentration visually. The images were used to calculate the intensity values using ImageJ software, which inturn was used to calculate the SGOT concentration. The PADs were also tested with serum samples, and SGOT spiked serum samples. The PAD could detect the SGOT concentration ranging from 5 to 200 U/L. The analysis yielded highly accurate results with less than 6% relative error compared to the clinical sample. This colourimetric test demonstrated exceptional selectivity in the presence of other biomolecules in the blood serum, with a detection limit of 2.77 U/L and a limit of quantification of 9.25 U/L. Additionally, a plasma separation membrane was integrated with the PAD to directly test SGOT from finger-prick blood samples.

Biosurfactant-capped CuO nanoparticles coated cotton/polypropylene fabrics toward antimicrobial textile applications.

The global COVID-19 pandemic has led to an increase in the importance of implementing effective measures to prevent the spread of microorganisms. Consequently, there is a growing demand for antimicrobial materials, specifically antimicrobial textiles and face masks, because of the surge in diseases caused by bacteria and viruses like SARS-CoV-2. Face masks that possess built-in antibacterial properties can rapidly deactivate microorganisms, enabling reuse and reducing the incidence of illnesses. Among the numerous types of inorganic nanomaterials, copper oxide nanoparticles (CuO NPs) have been identified as cost-effective and highly efficient antimicrobial agents for inactivating microbes. Furthermore, biosurfactants have recently been recognized for their potential antimicrobial effects, in addition to inorganic nanoparticles. Therefore, this research's primary focus is synthesizing biosurfactant-mediated CuO NPs, integrating them into natural and synthetic fabrics such as cotton and polypropylene and evaluating the resulting fabrics' antimicrobial activity. Using rhamnolipid (RL) as a biosurfactant and employing a hydrothermal method with a pH range of 9-11, RL-capped CuO NPs are synthesized (RL-CuO NPs). To assess their effectiveness against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) microorganisms, the RL-CuO NPs are subjected to antibacterial testing. The RL-capped CuO NPs exhibited antimicrobial activity at much lower concentrations than the individual RL, CuO. RL-CuO NPs have shown a minimum inhibitory concentration (MIC) of 1.2 mg ml-1and minimum bactericidal concentration (MBC) of 1.6 mg ml-1forE. coliand a MIC of 0.8 mg ml-1and a MBC of 1.2 mg ml-1forS. aureus, respectively. Furthermore, the developed RL-CuO NPs are incorporated into cotton and polypropylene fabrics using a screen-printing technique. Subsequently, the antimicrobial activity of the coated fabrics is evaluated, revealing that RL-CuO NPs coated fabrics exhibited remarkable antibacterial properties against both gram-positive and gram-negative bacteria.

A paper-based point-of-care testing device for the colourimetric estimation of bilirubin in blood sample.

A paper-based colourimetric assay for the Point-of-Care Testing (PoCT) of bilirubin has been developed based on the formation of a green-coloured copper-bilirubin complex from a blue-coloured tetraamminecopper(II) sulphate complex. The reaction was studied and optimized by UV-Visible absorption spectroscopy and translated onto a paper strip. Hydrophobic circular well patterns on Whatman chromatography paper were created by wax printing. The tetraamminecopper(II) sulphate complex was drop cast and dried on the reagent zones in the wax-patterned paper. The images of reagent zones captured using a scanner were analyzed using ImageJ software. Bilirubin spiked blood serum was tested in the concentration range of 1.2 to 950 µM. The PAD exhibited sensitivities of 0.4197 a.u/µM and 0.1040 a.u/µM for concentration ranges of bilirubin 1.2 to 96 µM and 105 to 950 µM respectively and a low detection limit of 0.799 µM. The method is highly selective to bilirubin, even in the presence of other biomarkers in serum. A plasma separation membrane incorporated PAD was fabricated for the final testing and quantification of bilirubin from whole blood.

Paper based micro/nanofluidics devices for biomedical applications.

This chapter details the significance, fabrication and biomedical applications of paper-based microfluidic devices. The first part of the chapter describes the importance of paper diagnostic devices, highlighting pretreatment, dipsticks, lateral flow assays, and microPADs. Various methods followed for the fabrication of the paper analytical devices are discussed in the second part. The last part is about some of the important biomedical applications of paper analytical devices. Finally, the challenges and research gaps in the paper microfluidics for biomedical applications are presented.

Development of a paper-based analytical device for the colourimetric detection of alanine transaminase and the application of deep learning for image analysis.

A paper-based colourimetric assay for the detection of alanine transaminase has been developed. In the presence of alanine transaminase, 2,4-dinitrophenyl hydrazine changes to pyruvate hydrazone leading to a colour change from pale yellow to dark yellow. Reaction conditions were optimized using absorption spectroscopic studies. Hydrophobic patterns on the Whatman chromatographic paper were created by wax printing, and the reagents were drop cast at the reagent zone. On the paper device, the intensity of the yellow colour increases with ALT concentration in the range of 20-140 U/L in human serum. For the quantification of ALT, coloured images were captured using a digital camera and were processed with Image J software. The machine learning approach was also explored for the ALT analysis by training with colour images of the paper device and testing using a cross-validation procedure. The results obtained with real clinical samples on the paper device showed good accuracy of less than 5% relative error with the clinical lab results. Furthermore, the paper device shows high selectivity to ALT in the presence of various interfering species in blood serum with a sensitivity of 0.261 a.u/(U/L), a detection limit of 4.12 U/L, and precise results with an RSD of less than 7%. For the testing of whole blood, a plasma separation membrane was integrated with the patterned paper.

Tantalum oxide honeycomb architectures for the development of a non-enzymatic glucose sensor with wide detection range.

Tantalum oxide honeycomb nanostructures (THNS) were fabricated by electrochemical anodisation of tantalum in H2SO4-HF medium. XRD analysis showed that annealing of THNS at 400 °C improves the crystallinity. HRSEM and AFM results illustrated that nanopores with an average diameter of 30 nm were uniformly distributed and the pore size reduced to 24 nm and 18 nm during subsequent electrodeposition of Pt and CuO. Electrodeposited Pt and CuO exhibited face centered cubic (fcc) and monoclinic crystal structure respectively. Cyclic voltammetric studies revealed that, on the hybrid material electrooxidation of glucose occurs at a lower potential (0.45 V). The sensor exhibited linear response to glucose up to 31 mM, fast response time (<3 s) and a low detection limit of 1 µM (S/N=3). The sensor is free of interference from ascorbic acid, uric acid, dopamine and acetaminophen. Sensor was used to analyze glucose in blood serum samples.