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Disposable Surgical Gloves and Pasteur (Transfer) Pipettes as Potential Sources of Contamination in Nitrite and Nitrate Assays

¹ Div. of Clin. Biochem., Depts. of Pediatric Lab. Med. and
² Cardiol., Critical Care Med. and Pediatrics, Univ. of Toronto, The Hosp. for Sick Children, 555 University Ave., Toronto, ON, M5G 1X8, and
³ Dept. of Clin. Biochem., Univ. of Toronto, 100 College St., Toronto ON, M5G 1L5, Canada;
^a address for correspondence: 17 Runnymede Rd., Toronto, ON, M6S 2Y1, Canada; E-mail graham.ellis@utoronto.ca

Much attention has been focused recently on the role of nitric oxide (NO) in physiology. NO has important effects as a bactericidal agent released by macrophages, as a relaxing factor causing vasodilatation, as a neurotransmitter, and as a paracrine substance (local "hormone") in many other physiological processes (1)(2). In humans, NO is rapidly converted to nitrite and nitrate, and these metabolites in blood or urine are often used to investigate its metabolism.

Disposable surgical gloves are needed in areas where specimens are collected, received, separated, and analyzed. Several types of gloves are available, in latex or latex-free formulation, with powder or powder-free. During the development of assays for nitrate and nitrite, we identified disposable gloves and glass Pasteur pipettes as potential sources of contamination.

We measured nitrite and nitrate with an HPLC method essentially following the method of Wennmalm et al. (3)(4). Water was 18 M (MilliQ water system, Millipore Canada). The solution to be tested was placed in a 1.0-mL disposable Wisp^TM vial (Waters Ltd., cat. no. WAT025054). Then 5 µL, diluted with water if necessary, was injected into a Waters HPLC system run isocratically at 1.0 mL/min with mobile phase of 10 mmol/L phosphate buffer, pH 8.0, a Waters IC-Pak Anion HR anion-exchange column [4.6 x 75 mm (cat. no. WAT026765)] with a Guard-Pak precolumn module containing IC-Pak Anion Guard-Pak Insert (cat. no. WAT010551). Spectrophotometric detection was at 214 nm. Samples were also tested for nitrite + nitrate (NOx) with the colorimetric Griess reaction as modified by Verdon et al. (5), after treatment of the sample with nitrate reductase from Aspergillus species (Boehringer Mannheim Canada, cat. no. 981249).

Several samples were also purified through Sep-Pak® Vacc 3cc (500 mg) tC₁₈ cartridges (Waters cat. no. WAT036815) to further confirm . . . [Full Text of this Article]

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References

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				B. Childress, J. K. Stechmiller, and G. S. Schultz Arginine Metabolites in Wound Fluids From Pressure Ulcers: A Pilot Study Biol Res Nurs, October 1, 2008; 10(2): 87 - 92. [Abstract] [PDF]