TOLUENE in Blood: METHOD 8007, Issue 1, dated 5 March 2013 - Page 2 of 5
REAGENTS:
EQUIPMENT:
1. Whole blood*, citrate treated 2. Toluene* (reagent grade, >99.5%) 3. Isobutanol internal standard (IS) as a 20 mg/L solution in water* 4. Water, ASTM Type II 5. Helium, UHP or higher 6. Hydrogen, UHP or higher 7. Air, UHP or higher 8. Toluene calibration standards* 9. Ethylene glycol*
- See SPECIAL PRECAUTIONS
1. Gas chromatograph, with FID, column, and data collector (page 8007-1) 2. Headspace sampler, holds up to 40 vials and can thermostat up to 12 vials simultaneously for automated headspace analysis (Any instrument that can achieve the headspace settings specified on p. 8007-1 is acceptable) 3. Bagged refrigerant, and refrigerator 4. Adjustable pipette, 10-mL, tips 10-mL, plastic or serological 5. Headspace vials, 20-mL, with caps and septa 6. Micro-liter syringes, 10-µL , 25-µL, 100-µL, 1-mL 7. Volumetric flasks, 10-mL, 25-mL, 1-L 8. Syringes, twist-on fitting (luer lock style or equivalent), 5-mL, and 20 gauge needles 9. Tubes, vacuum blood collection, 10-mL, citrate coated (light blue top)
SPECIAL PRECAUTIONS: Wear gloves, lab coat, and safety glasses while handling all chemicals and blood products. Disposable plastic, glass, and paper (pipet tips, gloves, etc.) that contact blood should be placed in a biohazard container. Contact with biological samples can have serious health consequences through exposure to hepatitis, HIV, and other diseases. All personnel collecting, handling, or analyzing samples should follow standard precautions [6] and comply with the OSHA bloodborne pathogens standard [7] which includes immunization for hepatitis B. Toluene is extremely flammable. All work should be performed in a fume hood. SAMPLING: 1. Collect the blood by filling a 10-mL vacuum blood collection tube (light blue top, citrate) as completely as possible in order to keep the air space to a minimum. Be certain to invert the blood tube several (5-10) times to mix the anticoagulant. NOTE: If desired, duplicate blood tubes could be drawn to allow for estimation of measurement precision, especially if collecting from numerous participants. 2. Immediately transfer the sample to a refrigerator or cooler (≤ 4 °C.) 3. Ship the sample vial in a well-insulated container equipped with blue ice or other cooling material. SAMPLE PREPARATION: 4. Allow the blood sample to reach room temperature, then mix gently so as to not produce a froth by inverting the tubes 4 to 5 times. 5. Add 1.5 mL of the IS solution to a 20-mL headspace vial. 6. Extract 1.5 mL of sample from the vacuum vial using a 5-mL syringe with a twist-on fitting (LuerLock style or equivalent) or micropipette and transfer the blood to a headspace vial. 7. Cap both sample and headspace vials immediately. 8. Mix the contents of each headspace vial thoroughly.
NIOSH Manual of Analytical Methods (NMAM), Fifth Edition
