Determination of ADME and Bioavailability Following Intravenous, Oral, and Dermal Routes of Exposure

Shakil A. Saghir1

1 Department of Biological and Biomedical Sciences, The Aga Khan University, Karachi, Pakistan
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 5.8
DOI:  10.1002/0471140856.tx0508s41
Online Posting Date:  August, 2009
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Abstract

Humans are exposed to chemicals either voluntarily or involuntarily through several routes. Therapeutic drugs are introduced into the human system via a number of routes including, but not limited to, oral, inhalation, intravenous (i.v.), topical, and subcutaneous. For occupational and environmental chemicals, the major routes of human exposure are inhalation, dermal, and oral. To determine the extent of exposure to chemicals, the concentration of the active molecules is measured in a biological medium. Determination of absolute and/or relative bioavailability of occupational and environmental chemical exposure through different routes is critical in understanding the risk to the general population of a low‐level exposure to these chemicals. This unit describes typical protocol designs to generate data for the calculation of absorption, distribution, metabolism, and elimination (ADME) and absolute and relative bioavailability of chemicals when exposed through i.v., oral, and dermal routes. Curr. Protoc. Toxicol. 41:5.8.1‐5.8.19. © 2009 by John Wiley & Sons, Inc.

Keywords: oral ADME; intravenous (IV) ADME; dermal ADME; bioavailability

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Conducting an Intravenous ADME Study
  • Support Protocol 1: Collection and Analysis of Plasma and Red Blood Cells
  • Support Protocol 2: Collection and Analysis of Urine
  • Support Protocol 3: Collection and Analysis of Feces
  • Support Protocol 4: Collection and Analysis of Expired Volatiles
  • Support Protocol 5: Collection and Analysis of Expired CO2
  • Alternate Protocol 1: Conducting an Oral ADME Study
  • Support Protocol 6: Determining Oral Absorption
  • Alternate Protocol 2: Conducting a Dermal ADME Study
  • Support Protocol 7: Sample Collection by Dermal Wash
  • Support Protocol 8: Sample Collection from Dermal Application Devices
  • Support Protocol 9: Sample Collection by Tape Stripping of the Dermal Application Site
  • Support Protocol 10: Sample Collection of the Dermal Application Site
  • Support Protocol 11: Determining Dermal Absorption
  • Support Protocol 12: 14C Analysis of Samples
  • Support Protocol 13: Collection and Analysis of Tissues
  • Support Protocol 14: Collection and Analysis of Final Cage Wash
  • Support Protocol 15: Metabolic Profile and Identification
  • Support Protocol 16: Collection of Control Samples
  • Basic Protocol 2: Data Analysis
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Conducting an Intravenous ADME Study

  Materials
  • Saline or a balanced salt solution (BSS)
  • 10% Intralipid (lipid emulsion; Fresenius Kabi or prepared as described by Saghir et al., ) or other suitable aqueous miscible solvent/emulsifier
  • 1 to 5 N HCl or NaOH
  • 14C‐ or 3H‐radiolabeled test compound (ideally ≥97% pure)
  • Non‐radiolabeled test compound (≥97% pure), if necessary
  • Scintillation cocktail
  • Jugular vein–cannulated (JVC) rats (Fig. ; at least 8 weeks old; Taconic)
  • Rodent chow in pellet form
  • Cannula flush solution (see recipe)
  • Cannula lock solution (see recipe)
  • Isopropyl alcohol
  • Silicone grease
  • Activated charcoal
  • 3:7 (v/v) monoethanolamine/1‐methoxy‐2‐propanol solution
  • Scintillation counter
  • Glass rat metabolism cages that separate urine and feces and collect exhaled air (see Fig. for detailed setup)
  • 1‐ to 3‐ml Luer‐lok glass syringe
  • Luer‐stub adapter (23‐G) with blunt end
  • Forceps and hemostats
  • Warming lamp with 40‐ to 100‐W bulb or ∼45°C incubator equipped with mechanical convection
  • Hard Plexiglas rat holder or preferably cone‐shaped flexible plastic bag (e.g., Decapicone, Braintree Scientific)
  • Board and tape
  • 0.5‐in. long, precision glide 23‐G needle with short beveled tip
  • 0.5‐in. long blunt‐end Luer‐stub adapter (23‐G)
  • 1‐ or 2‐ml Luer‐lok glass syringe
  • Cotton pledget or gauze
  • Additional reagents and equipment for sample collection (see Support Protocols protocol 21 through protocol 65)

Support Protocol 1: Collection and Analysis of Plasma and Red Blood Cells

  • Ball‐tipped fixed or flexible feeding needle (16‐ to 18‐G curved or straight)

Support Protocol 2: Collection and Analysis of Urine

  • Isoflurane
  • Electric hair clippers
  • Teflon frames (∼1.5‐mm thick; 4 × 5–cm with a 3 × 4–cm cut‐out opening)
  • Adhesive Velcro fastener
  • Rodent jackets (Braintree Scientific)
  • Ball‐tipped feeding needle (∼20‐G) and all‐glass Luer‐lok syringe
  • Teflon spectra/mesh macroporous filter (Spectrum Laboratories)
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Figures

Videos

Literature Cited

Literature Cited
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   U.S. EPA (United States Environmental Protection Agency) 1996. Health effects test guidelines OPPTS 870.7600: Dermal penetration. Prevention, Pesticides and Toxic Substances (7101). Office of Pesticide Programs, Washington, D.C., report 712–C–96–350.
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