Ultraviolet spectrophotometer

The action of biochemical parameters such as serum alanine transaminase (ALT), alkaline phosphate (ALP), bilirubin, total cholesterol (CHL), albumin (ALB), and serum lactate dehydrogenase (LDH) was determined using standard kit methods. All estimation was done using ultraviolet spectrophotometer (Shimadzu) as per standard kit methods.

Total phenolic content was measured using the Folin–Ciocalteu’s reagent as described [11 ]. Absorbance was measured at 760 nm using an ultraviolet spectrophotometer (Shimadzu, Japan). Gallic acid was used as a standard. A dose response linear regression was generated using the gallic acid standard absorbance, and the total phenolic content was expressed as mg gallic acid equivalents (GAE)/g dry weight of extract. Values were determined in triplicate.

It was used as a thiobarbituric acid reacting substances (TBARS) assay. It was performed as described by Jentzsch et al.^{18 (link)} In the TBARS assay, one molecule of malondialdehyde (MDA) reacts with 2 molecules of thiobarbituric acid (TBA) to produce a pink pigment with absorption at 535 nm. Amplification of peroxidation during the assay is prevented by the addition of the chain-breaking antioxidant Butylated hydoxytoluene (BHT). Plasma (400 μL) or MDA standard (0.2-4 μmol/L) was prepared by hydrolysis of 1,1,3,3 tetramethoxypropane (Sigma Chemical Co, St Louis, Missouri) and was mixed with 400 μL orthophosphoric acid (0.2 mol/L) (Sigma Chemical Co) and 50 μL BHT (2 mmol/L) (Sigma Chemical Co) in 12 × 75 mm tubes. We added 50 μL TBA reagent (0.11 mol/L in 0.1 mol/L NaOH) (Fluka Chem., Buchs, Switzerland) and mixed the contents; the contents were then incubated in a water bath at 90°C for 45 min. The tubes were put on ice to stop the reaction. Thiobarbituric acid reacting substances were extracted once with 1000 mL n-butanol (Sigma Chemical Co, St. Louis, MO, USA). The upper butanol phase was read at 535 nm and 572 nm to correct for baseline absorption using an ultra violet spectrophotometer (Shimadzu Corporation, Japan). MDA equivalents (TBARS) were calculated using the difference in absorption at the 2 wavelengths, and quantification was done using the calibration curve.¹¹

Phytovesicles containing AFTSI were separated from untrapped drug by centrifugation at 9000 rpm for 45 min. The supernatant was recovered and assayed spectrophotometrically using Shimadzu ultraviolet spectrophotometer.
Where, Ct - concentration of total AFTSI; Cr - concentration of free AFTSI.