Misr International University

An accessible eco-friendly chromatographic methodology was established by integrating flow injection analysis (FIA) with on-line solid phase extraction (SPE) into a conventional high performance liquid chromatography (HPLC) equipped with fluorescence detection (FLD) (FIA-SPE-HPLC-FLD) for quantifying methotrexate (MTX) in untreated serum samples. The FIA-SPE-HPLC-FLD method incorporated a cerium (IV) trihydroxy hydroperoxide (CTHPO) packed column for the derivatization of MTX into highly fluorescent derivatives. This step was followed by on-line SPE of matrix proteins with a protein-coated C8 column (PC-C8-column). For the CTHPO-column, a carrier stream of potassium dihydrogen phosphate (0.01 M, pH 3.4) was used, which also functioned as the extraction mobile phase for PC-C8-column. Automated SPE relied on the principle of size exclusion liquid chromatography performed under non-denaturing conditions. In measuring stage, the fluorescent derivatives were transferred to the TSK gel ODS-80 TM, analytical column, by employing a mixture of acetonitrile and potassium dihydrogen phosphate (0.01 M, pH 6.0) in a 10:90 (v:v) ratio. Quantification was conducted using excitation and emission wavelengths of 367 nm and 463 nm, respectively. Both the CTHPO- andPC-C8-columns exhibited substantial effectiveness in chemically modifying the analyte and extracting proteins. Validation of the FIA-SPE-HPLC-FLD method demonstrated good linearity across a concentration range of 5-1000 ng/mL MTX, with recovery percentages between 96.86 % and 98.15 %, indicating minimal matrix effect. The FIA-SPE-HPLC-FLD method aligns with 9 out of the 10 principles of GSP, highlighting its sustainability for evaluating lower concentrations of MTX in clinical human serum samples.

The present novel trial assesses the prophylactic influence of ZnO NPs in comparison to silymarin against liver damage induced by acetaminophen (APAP). Forty albino rats were allocated into 4 groups (n = `10 rats/ group). Group I (Control), was orally administered 0.9 % NaCl for 21 days. Group II (exposed to APAP) received distilled water (1 ml/kg per day) via oral gavage for 19 successive days followed by APAP (600 mg/kg, PO) twice daily for 2 days. Group III (APAP+ Silymarin) was orally administered silymarin at the dose of 200 mg/kg daily for 19 days followed by APAP (600 mg/kg) via oral gavage twice daily for 2 successive days. Group IV (APAP+ ZnO NPs) was orally given ZnO NPs at the dose of 100 mg/kg daily for 19 days followed by (600 mg/kg/ twice daily) APAP orally for 2 successive days. APAP exhibited a substantial elevation of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, nitric oxide and malondialdehyde levels. Meanwhile, APAP markedly decreased catalase, SOD, GST and TAC levels. A significant elevation in DNA damage inside hepatocytes was noticed. APAP induced many histopathological changes. Conversely, the prophylactic use of ZnO NPs inhibited the elevation of AST, ALT, ALP, NO and MDA as well as the decline of catalase, SOD, GST and TAC levels induced by APAP toxicity compared to group III. Marked improvement of the architecture of hepatic tissue was noticed in group IV. In conclusion, ZnO-NPs were more actual in ameliorating APAP hepatotoxicity compared to silymarin via antioxidant and antiapoptotic pathways.