Brilliant green

Guidelines (The recommended experimental protocol below is for guidance only and should be adjusted according to your specific requirements)

1. Stock Solution Preparation
1.1 Solvent: Water or 60% ethanol. Aqueous solutions are unstable and require fresh preparation.
1.2 Concentration recommendation: It is generally recommended to prepare a high-concentration stock solution of 1-10 mM.

2. Working Solution Preparation
2.1 Diluent: Double-distilled water.
2.2 Working concentrations: 1% (based on 60% ethanol), 0.5% ethanol solution, 1:2000 (aqueous solution), 1:1000 (aqueous solution).
2.3 Notes: Adjust the working solution concentration as needed; prepare fresh before use.

3. Staining Procedures
3.1 Sample type description
3.1.1 Bacterial samples: Suitable for Gram-positive bacteria, with weak efficacy against *Escherichia coli* and Mycobacterium tuberculosis.
3.1.2 Skin/mucosa: 1% ethanol solution is used for skin disinfection; 0.5% solution is used for mucosa.
3.1.3 Adsorption experiment samples: Add 25 mg of PANI/Ag nanocomposite to 20 mL of BG solution (initial concentration 40-61.2 mg/L), shake at 30 °C and 120 rpm, and reach adsorption equilibrium within 120 min.
3.2 Incubation conditions
3.2.1 Antibacterial experiment: Apply to skin, mucosa or wounds at room temperature or body temperature; use warm aqueous solution for wet compresses.
3.2.2 Adsorption experiment: Shake at 30 °C and 120 rpm in the dark, with a contact time of 120 min.
3.3 Washing steps
3.3.1 Skin staining can be removed by vigorous wiping with alcohol or hydrogen peroxide.

4. Control Setup
4.2 Adsorption experiment controls: Comparison of adsorption efficiency under conditions of different initial concentrations (40-61.2 mg/L), different nanocomposite dosages (0.01-0.03 g), different temperatures (25-40 °C) and different pH values (2-12).

5. Detection and Analysis
5.1 Instrument type: Spectrophotometer.
5.2 Excitation/emission wavelength: 625 nm (detection wavelength).
5.3 Result analysis
5.3.1 Absorbance change: The absorbance at 625 nm decreases with the extension of adsorption time; the removal rate reaches 89.8% at an initial concentration of 40 mg/L and 75% at 61.2 mg/L; approximately 90% is removed within 120 min.
5.3.2 Antibacterial activity: The minimum inhibitory dilution against streptococci and pneumococci is 1:200,000, with weak efficacy against *Escherichia coli* and Mycobacterium tuberculosis.
5.3.3 Color change: Green dye. Adsorption is the lowest at pH 2; the removal rate increases from 61% to 85% at pH 4-12, reaching the maximum at pH 12; the pKa values are 4.93 and 2.26.