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Rapid CommunicationAccelerated Communication

Comparison of In Vivo and In VitroDeposition of Rhodamine and Fluorescein in Hair

Peter R. Stout and James A. Ruth
Drug Metabolism and Disposition October 1998, 26 (10) 943-948;
Peter R. Stout
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James A. Ruth
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Abstract

A direct differentiation of the internal and external drug-deposition pattern into hair was made using two fluorescent dyes and fluorescence microscopy after systemic administration to mice or external exposure of untreated hair. Mice (23 days old, C57 and Balb/C) were administered either rhodamine or fluorescein intraperitoneally at varied doses on 3 consecutive days of 3 weeks, and hair was sampled 1 week later. Another group was given 10 mg/kg rhodamine or 100 mg/kg fluorescein and sampled at time points from 5 min to 168 hr. The time courses of external deposition of rhodamine and fluorescein into untreated hair were examined after hair was soaked in 0.1 mg/ml solutions at pH 3, pH 6, and pH 9 aqueous buffer or methanol. The hair was then extracted in pH 6 phosphate buffer or methanol for 24 hr.In vivo accumulation was distinguishable as fluorescent bands along the length of the hair for rhodamine and fluorescein. The pattern of in vivo deposition appears to arise from the rapid accumulation within the cortex and medulla, with little deposition evident in the cuticle. Neither phosphate buffer nor methanol washes affected the intensity of fluorescence in the hair. External loading of rhodamine into the hair resulted in staining of the junctions of cuticle scales. This pattern persisted even after 12 hr of solution exposure. Extraction with pH 6 phosphate buffer or methanol did not remove rhodamine. Fluoroscein followed a similar pattern, with maximum fluorescence when hair was loaded in pH 6 100mM phosphate buffer and nominal staining when loaded in pH 9 100 mM Tris buffer or methanol. Soaking the hair in pH 6 buffer, but not methanol, removed some fluorescein. These results demonstrate that compounds in the circulation can rapidly diffuse into the forming cortex and medulla, where rapid associations occur with elongating intermediate filaments specific to the medulla and cortex. These compounds can become significantly occluded within the mature matrix and are resistant to removal in aqueous or methanolic solutions.

Footnotes

  • Send reprint requests to: James A. Ruth, Ph.D., University of Colorado Health Sciences Center, Department of Molecular Toxicology and Environmental Health Sciences, 4200 East. Ninth Avenue, Box C238, Denver, CO 80262.

  • This work supported by NIH grant DA09545.

  • Abbreviations used are::
    IFs
    intermediate filaments
    IRS
    internal root sheath
    CMC
    cell membrane complex
    ERS
    external root sheath
    • Received July 20, 1998.
    • Accepted August 17, 1998.
  • The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition
Vol. 26, Issue 10
1 Oct 1998
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Rapid CommunicationAccelerated Communication

Comparison of In Vivo and In VitroDeposition of Rhodamine and Fluorescein in Hair

Peter R. Stout and James A. Ruth
Drug Metabolism and Disposition October 1, 1998, 26 (10) 943-948;

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Rapid CommunicationAccelerated Communication

Comparison of In Vivo and In VitroDeposition of Rhodamine and Fluorescein in Hair

Peter R. Stout and James A. Ruth
Drug Metabolism and Disposition October 1, 1998, 26 (10) 943-948;
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