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9 protocols using bilirubin

1

Acetaminophen Toxicity Assessment Protocol

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Acetaminophen was a product of Santa Cruz Biotechnology (Santa Cruz, CA). Diphenylamine, 5,5′-dithio-bis(2-nitrobenzoic acid), guanidine hydrochloride and N-ethyl-maleimide (NEM) were procured from Research Organics (Cleveland, OH). Alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, bilirubin, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Red) and glucose 6-phosphate dehydrogenase (Glc-6-PD) were products of Randox Laboratories Ltd., Co. (Antrim, UK). All other reagents used were supplied by Sigma-Aldrich Inc. (St. Louis, MO).
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2

Albino Rat Biochemical Assays

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Twenty-four albino male rats (Ratus novergicus) with an average weight of 140 g used for this study were purchased from the Animal Holding Unit of the Department of Biochemistry, University of Ibadan, Nigeria. ATR-MET is a product of First Chemical Co Ltd, Changxing, Zhejiang China. GSH, 1-chloro-2, 4-dinitrobenzene (CDNB), 5′, 5′-dithio-bis-2-nitrobenzoic acid (DTNB), thiobarbituric acid (TBA), epinephrine and hydrogen peroxide were purchased from Sigma Chemical Company (London, UK). ALT, AST, ALP, GGT, urea, creatinine, bilirubin, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides kits were obtained from Randox Laboratories Ltd, Antrim UK. All other chemicals and reagent were of analytical grade and were obtained from British Drug House, Poole, UK.
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3

Biochemical Assays for Liver and Kidney Evaluation

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All chemicals and reagents were of analytical grade and were used as supplied. Assay kits for alkaline phosphate (ALP), aspartate transaminase (AST), alanine transaminase (ALT), creatinine (CREA), urea, albumin (ALB), and bilirubin (BIL) were products of Randox Laboratories Limited (Crumlin, UK).
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4

Nanoparticle-based Biochemical Assay Protocol

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Magnesium hydroxide (Mg(OH) 2 nanoparticles, 99% purity, 10nm size) were obtained from US Research Nanomaterials, Inc. 3302 Twig Leaf Lane Houston, TX 77084, USA. The assay kits for creatinine, calcium, sodium, potassium, chloride, magnesium, albumin, bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT) alkaline phosphatase (ALP), acid phosphatase (ACP), were purchased from Randox Laboratories United Kingdom. All other reagents used were of analytical grade.
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5

Moxifloxacin-Induced Oxidative Stress

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Moxifloxacin hydrochloride (Avelox) was a product of Bayer Healthcare Pharmaceutics, Leverkusen, Germany. Glutathione (GSH), 1-chloro-2,4-dinitrobenzene (CDNB), 5′,5′-dithiobis-2-nitrobenzoic acid (DTNB), thiobarbituric acid (TBA), epinephrine, and hydrogen peroxide were purchased from Sigma Chemical Company (London, UK). Assay kits for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyl transferase (γ-GT), urea, creatinine, bilirubin, total cholesterol, HDL-cholesterol, LDL-cholesterol, and triglycerides were products of Randox Laboratories Ltd., Antrim, UK. All other chemicals and reagents used in this study were of analytical grade.
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6

Phytochemical and Biochemical Analysis

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Methanol, acetic acid, quercetin, and gallic acid were purchased from Merck (Darmstadt, Germany). Sodium carbonate (Na2CO3), aluminum chloride, potassium acetate, and Folin-Ciocalteu’s reagent were procured from Sigma Aldrich, Inc (St Louis, MO). Isobutyl alcohol, magnesium carbonate (MgCO3), iron (III) chloride (FeCl3), acetone, chloroform, sodium nitroprusside, Methanol, and ferric chloride were source from BDH Chemical Ltd (Poole, England). Water used was glass distilled. Creatinine (Lot No. 315822), aspartate aminotransferase (AST; Lot No. 295800), alanine aminotransferase (ALT; Lot No. 295830), bilirubin (Lot No. 311158), uric acid (UA; Lot No. 309349), total protein (Lot No. 261368), and albumin (Lot No. 302605) kits were sourced from RANDOX Laboratories Ltd, Crumlin, Co, Antrim, UK. Kenxin refrigerated centrifuge model KX3400C was used, while a Jenway UV-visible spectrophotometer (Model 6305; Jenway, Barlo World Scientific, Dunmow, UK) was used to measure the absorbance.
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7

Haloxyfop-p-methyl ester Biochemical Assay

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Haloxyfop-p-methyl ester (Gallant Super®) was purchased from Zhejiang Jinfanda Biochemical Co., Ltd., Zhejiang, China. Glutathione, 1-chloro-2,4-dinitrobenzene(CDNB), 5,5′-dithio-bis-2-nitrobenzoic acid (DTNB), epinephrine and hydrogen peroxide (H2O2) were obtained from Sigma® Chemical Company, London, U.K. Assay kits for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), acid phosphatase (ACP), gamma glutamyl transferase (γ-GT), urea, creatinine and bilirubin were products of RANDOX® Laboratories Ltd., Antrim, U.K. The assay kit for lactate dehydrogenase (LDH) was obtained from Cypress Diagnostics, Langdorp, Belgium. All other reagents employed were of analytical grade and highest purity.
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8

Capecitabine Hepatotoxicity Evaluation

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The substances used include the following: Capecitabine tablets (Xeloda®), a product of Rochie Product Ltd., Castle Hill, Australia; Glutathione (GSH), 1-chloro-2, 4-dinitrobenzene (CDNB), 5’,5’-dithio-bis-2-nitrobenzoic acid (DTNB), thiobarbituric acid (TBA), epinephrine and hydrogen peroxide were purchased from Sigma® Chemical Company (London, UK); alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), Urea, creatinine and bilirubin kits were obtained from RANDOX® Laboratories Ltd., Antrim, UK. All other chemicals and reagent were of analytical grade and were obtained from British Drug House, Poole, UK.
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9

Hepatotoxicity Evaluation Protocol

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Acetaminophen, chloroform, and methanol used were obtained from Fam-lab Nigeria Limited, Akure, and Lixok-k chemicals, Akure, respectively. Alanine transaminase (ALT), aspartate transaminase (AST), gamma glutamyltransferase (GGT) and total protein, and bilirubin were obtained from Randox Laboratories Limited, UK.
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