Lc 321 322 350

3-indoxyl sulfate (a metabolite of tryptophan amino acid) is well-known uremic toxin that enhances renal injury via glomerular sclerosis and interstitial fibrosis. 3-indoxyl sulfate is also involved in reactive oxygen species (ROS) production in kidney cells, thus increasing oxidative stress. The amount of 3-IS in urine, serum, and kidney tissue was analyzed by high-performance liquid chromatography (HPLC). Organic solvent was used for the extraction of samples using a multistep process. After extraction, the samples were mixed with an internal standard (2-napthalenesulfonic acid) thoroughly and analyzed by HPLC (Gilson, LC-321 322 350) at 280 nm. The extraction recovery of samples was found to be 87%.

The 3-IS content was estimated in the serum, urine, and kidney tissues using high-performance liquid chromatography (HPLC). The extraction of the samples was performed using 70% acetonitrile, an organic solvent, in a multi-step process. The extracted samples were mixed thoroughly with 2-naphthalene sulfonic acid used as an internal standard, and measured using HPLC (Gilson, LC-321322350) at 280 nm. The retention time of 3-IS was 6.8 min, and the extraction recovery was 84%. The HPLC apparatus used in the present study comprised a Gilson pump (LC-321322350 pump), an autosampler (Gilson-234), and a UV (UV/VIS-151) detector (Gilson, France). A C18 column from Agilent (Torrance, CA, USA) with a pre-column (250 mm 4.6 mm, and 5 m) was used for detection and quantification with a flow rate of 0.9 mL/min at room temperature. A 70:30 v/v combination of acetonitrile and 0.1% trifluoroacetic acid in Milli Q water was employed as the isocratic mobile phase.