The aim of this study was to investigate the surface variability of 13 powder-free, unlined, and unsupported nitrile rubber gloves using attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectrophotometry at key wavelengths for analysis of captan contamination. The within-glove, within-lot, and between-lot variability was measured at 740, 1124, 1252, and 1735 cm<sup>−1</sup>, the characteristic captan reflectance minima wavelengths. Three glove brands were assessed after conditioning overnight at relative humidity (RH) values ranging from 2 ± 1 to 87 ± 4% and temperatures ranging from −8.6 ± 0.7 to 59.2 ± 0.9 °C. For all gloves, 1735 cm<sup>−1</sup> provided the lowest background absorbance and greatest potential sensitivity for captan analysis on the outer glove surface: absorbances ranged from 0.0074 ± 0.0005 (Microflex) to 0.0195 ± 0.0024 (SafeSkin); average within-glove coefficients of variation (CV) ranged from 2.7% (Best, range 0.9–5.3%) to 10% (SafeSkin, 1.2–17%); within-glove CVs greater than 10% were for one brand (SafeSkin); within-lot CVs ranged from 2.8% (Best N-Dex) to 28% (SafeSkin Blue); and between-lot variation was statistically significant (<i>p</i> ≤ 0.05) for all but two SafeSkin lots. The RH had variable effects dependent on wavelength, being minimal at 1735, 1252, and 1124 cm<sup>−1</sup> and highest at 3430 cm<sup>−1</sup> (O–H stretch region). There was no significant effect of temperature conditioning. Substantial within-glove, within-lot, and between-lot variability was observed. Thus, surface analysis using ATR-FT-IR must treat glove brands and lots as different. ATR-FT-IR proved to be a useful real-time analytical tool for measuring glove variability, detecting surface humidity effects, and choosing selective and sensitive wavelengths for analysis of nonvolatile surface contaminants.

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