Page:NIOSH Manual of Analytical Methods - 8303.pdf/2

 PENTACHLOROPHENOL in urine: METHOD 8303, Issue 2, dated 15 August 1994 - Page 2 of 4 REAGENTS:

EQUIPMENT:

1. Hexane, acetone, and benzene* (pesticide grade). 2. 20% benzene in hexane (v/v). 3. 10% benzene in hexane (v/v). 4. Hydrochloric acid (conc.). 5. Sodium bisulfite. 6. Sodium sulfate. 7. Alumina (acid-washed), Brockman Activity I, 80/200 mesh. 8. Pentachloroanisole (available from EPA Analytical Reference Lab). 9. Calibration stock solution equivalent to 100 mg/L PCP. Weigh 0.0105 g pentachloroanisole; dilute to 100 mL in a volumetric flask with hexane. Stable 2 months if stored in refrigerator. 10. Diazomethane*. Prepared from Diazald kit according to manufacturer's instructions [4]. 11. 5% methane in argon.

See Special Precautions.

1. Bottles, polyethylene, 125-mL, screw cap.** 2. Gas chromatograph with 63Ni electron capture detector. 3. Culture tubes, 16 x 150-mm and 16 x 125mm, with PTFE-lined screw caps.** 4. Mixer, rotary, variable speed. 5. Centrifuge (refrigerated, optional). 6. Chromatography column, 7-mm ID x 200 mm, with 50-mL reservoir and PTFE stopcock.** 7. Syringes, 10-µL, glass. 8. Pipets, 1000-, 500-, 250-, 100-, 50-, 25- and 10-µL.** 9. Flasks, volumetric, 10- and 100-mL.** 10. Pipets, Pasteur, disposable. 11. Tubes, centrifuge, graduated, glass-stoppered, 15-mL.** 12. Waterbath, 100 °C.

Wash with detergent; clean with chromic acid; rinse with distilled water, acetone, and hexane.

SPECIAL PRECAUTIONS: Samples of urine collected from humans pose a real health risk to laboratory workers who collect and handle these samples. These risks are primarily due to personal contact with infective biological samples and can have serious health consequences, such as infectious hepatitis, and other diseases. There is also some risk from the chemical content of these samples, but this is much less. Those who handle urine specimens should wear protective gloves, and avoid aerosolization of the samples. Mouth pipetting, of course, must be avoided. Benzene is a documented human carcinogen and must be handled in compliance with 29 CFR 1910.1005. Diazomethane and its precursor, N-methyl- N-nitroso- p-toluene sulfonamide are potent mutagens. Handle the bulk precursor in a glove box. Generate diazomethane behind an explosion shield in a hood.

SAMPLING: 1. 2. 3.

Collect a spot urine sample of 100 mL in a 125-mL polyethylene bottle. Add two or three drops conc. HCl to the sample as a preservative. Freeze the samples and pack them in an insulated shipper (e.g., styrofoam with dry ice).

SAMPLE PREPARATION: 4. 5. 6. 7. 8. 9. 10. 11.

Perform a creatinine determination on an aliquot of the urine (e.g., [5]). Transfer a 4-mL aliquot of each urine sample to a 16 x 150-mm culture tube. Add 1.0 mL conc. HCl and 100 mg sodium bisulfite to each culture tube. Cap and place the culture tubes in a boiling waterbath for 1 h. Shake gently at 15-min intervals. Cool the culture tubes to room temperature. Add 5 mL benzene to each culture tube. Extract on a rotary mixer at 60 rpm for 1 h. Centrifuge at 3000 rpm. Transfer benzene layer to a 16 x 125-mm culture tube. Repeat the extraction with an additional 5 mL benzene. Combine extracts. Concentrate the extract to approximately 0.6 mL under a gentle stream of nitrogen. NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94