April 1993
Volume 34, Issue 5
Free
Articles  |   April 1993
Quantitation of total protein deposits on contact lenses by means of amino acid analysis.
Author Affiliations
  • G Yan
    Department of Bioengineering, University of Utah, Salt Lake City 84112.
  • G Nyquist
    Department of Bioengineering, University of Utah, Salt Lake City 84112.
  • K D Caldwell
    Department of Bioengineering, University of Utah, Salt Lake City 84112.
  • R Payor
    Department of Bioengineering, University of Utah, Salt Lake City 84112.
  • E C McCraw
    Department of Bioengineering, University of Utah, Salt Lake City 84112.
Investigative Ophthalmology & Visual Science April 1993, Vol.34, 1804-1813. doi:
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    • Get Citation

      G Yan, G Nyquist, K D Caldwell, R Payor, E C McCraw; Quantitation of total protein deposits on contact lenses by means of amino acid analysis.. Invest. Ophthalmol. Vis. Sci. 1993;34(5):1804-1813.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: This study was done to characterize and quantify the protein deposits on worn contact lenses and to measure the residual deposits after extraction in 2% sodium dodecyl sulfate and the total protein deposits on worn vifilcon, atlafilcon, and tefilcon lenses (Food and Drug Administration Types IV, II, and I, respectively). METHODS: Contact lens extracts were separated with gel electrophoresis, and the amount of protein was estimated after silver staining and densitometry. To determine the residual deposits, the contact lenses were hydrolyzed, and amino acid analysis was carried out by reverse-phase high-performance liquid chromatography after precolumn derivatization with phenylisothiocyanate. Refinement of the hydrolysis conditions was undertaken to minimize interference by the lens polymers. RESULTS: The extraction removed only approximately 25% of the protein deposits. Mild hydrolytic conditions, 20 hr in 6 N HCl at 105 degrees C, were found to cause minimal polymer interference. Of the 350, 10, and 20 micrograms of protein typically determined on whole vifilcon, atlafilcon, and tefilcon lenses, the polymers were estimated to account for 4, 0.5, and less than 0.4 micrograms, respectively. CONCLUSIONS: Hydrolysis of worn contact lenses with subsequent amino acid separation can be applied to determine the total protein deposits without the uncertainty inherent in extraction of the deposits.

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