Page:NIOSH Manual of Analytical Methods - 8001.pdf/2

 PENTACHLOROPHENOL IN BLOOD: METHOD 8001, Issue 2, dated 15 August 1994 - Page 2 of 4 EQUIPMENT:

REAGENTS: 1.

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



See SPECIAL PRECAUTIONS.

1. Blood-sampling tubes, 5-ml, with 7.5 mg Na 2EDTA anticoagulant (e.g., "lavender-top" tubes). 2. Gas chromatograph with a 63Ni electron capture detector, electronic integrator, and column (page 8001-1). 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 a 50-mL reservoir and PTFE stopcock.** 7. Syringe, 10-µL, glass. 8. Pipets, 10- to 1000-µL.** 9. Flasks, volumetric, 10-, 25-, and 100-mL.** 10. Pipets, Pasteur, disposable. 11. Tubes, centrifuge, graduated, glass-stoppered, 15-mL.** 12. Waterbath, 100 °C. acid; rinse with distilled water, acetone, and hexane.
 * Wash with detergent; clean with chromic

SPECIAL PRECAUTIONS: Samples of blood 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 blood 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.

Collect 5-mL whole blood samples by venipuncture in tubes containing 7.5 mg Na 2EDTA anticoagulant, about 4 h after suspected exposure. Ship the blood samples in an insulated container with refrigerant to keep the temperature at about 10 °C. Do not freeze.

SAMPLE PREPARATION: 3. 4. 5. 6.

Add 2 mL whole blood to a 16 x 150-mm culture tube. Add cautiously and slowly 5 mL 6 M sulfuric acid. Cool to room temperature. Add 6 mL hexane to the culture tubes in a hood; cap and place in a boiling waterbath for 45 min with shaking every 15 min. Cool the culture tubes to room temperature and centrifuge at 2000 to 2500 rpm. Remove the hexane layer into a 16 x 125-mm culture tube. NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, 8/15/94