DEMOCOPHES Train the trainers Training module 1: Fieldwork

Transcription

1 DEMOCOPHES Train the trainers Training module 1: Fieldwork PREANALYTICAL PHASE 1

2 WP3 Sample handling, analysis and biobanking Leader: ARGELIA CASTAÑO Argelia Castaño José Antonio Jiménez Marta Esteban Jürgen Angerer Holger M. Koch Birgit Schindler

3 General aspects of the preanalytical phase DEMOCOPHES scenario Basic information on hair and urine Material: selection and washing of vessels Packing and shipment Samples reception Aliquoting and storage

4 Absorption Sedimentation Chemical changes Evaporation [analyte]

5 DEMOCOPHES SCENARIO BIOLOGICAL MATRICES Scalp hair and urine BIOMARKERS mercury cadmium cotinine phthalates creatinine BPA parabenes triclosan

6 HAIR 1 cm/month Main factors affecting hair growth: - Type - Body location - Age - Sex - Race Growth in cycles of growing and quiescence Applications: forensic and clinical toxicology, occupational medicine, doping control or HBM.

7 HAIR

8 HAIR CONSIDERATIONS: - Exposure and time period - Type of substance and metabolism - Clinical relevance CONCLUSIONS: 1. Recent exposure, > 1 year NOT USEFUL 2. Use of SOPs is essential 3. Supportive tool 4. Exceptions: As and Hg

9 HAIR FISH: 75-95% of Hg is MeHg ABSORPTION: 95 % of MeHg HAIR: Hg and MeHg levels linearly correlated; 70-80% of Hg is MeHg Global Mercury Assessment, 2002

10 URINE Widely used High number of biomarkers Metabolites Non-invasive, easy collection, no volume limitation

11 URINE 24 hr samples Spot samples Cumbersome Volume variations Improper/incomplete collection CREATININE ADJUSTMENT WHO exclusion criteria: [creatinine] < 0.3 g/l [creatinine] > 3.0 g/l

12 URINE CONTAMINATION Parent chemical Metabolite Risk Cadmium BPA Triclosan Parabenes Cotinine Phthalates metabolites Creatinine

13 URINE: Sampling vessels PTHALATES! BPA! SARSTEDT Ref ml, polypropilene SCHARLAB Ref ml, polypropilene PETE: polyethylene terephthalate HDPE: high density polyethylene PVC: polyvinyl chloride LDPE: low density polyethylene PP: polypropylene PS: polystyrene OTHER: polycarbonate, epoxy resins

14 URINE: pre-treatment of vessels

15 1. Prepare a 10% nitric acid solution from nitric acid 65% extra pure and purified water. 240 vessels 18 L HNO 3 solution 2.8 L HNO 3 65% 15.2 L Milli-Q water

16 2. Open the vessels and put vessels and lids into the tank. Overnight / 3 hours COMPLETELY IMMERSED!

17 3. Take out the vessels from the acid tank and put them in a first tank with purified water. Then, change the vessels and lids to the second tank of purified water. Acid solution Tank 2 Tank 1

18 4. Take out vessels and lids and put them face down in a clean filter paper in order to dry them.

19 5. Once the drying is finished, lids are screwed to the resulting nitric acid pre-treated vessels.

20 5% control material

21 URINE SAMPLING - Washing vessels - First-morning urine (or 5 hours) COPHES D.3.1 Instruction for the volunteers

22 PACKING AND SAMPLES SHIPMENT

23 Packing required: (examples from FedEx instructions) 1. Watertight primary receptacles 2. Watertight secondary receptacles

24 3. Absorbent material 4. Sturdy outer packing

25 Absorbent material and secondary container Outer packing 2 UN3373 bags with absorbent material + 1 bag for documents 4 cooling agents 2 box-racks Primary container

26

27 SAMPLES RECEPTION

28 ALIQUOTING AND STORAGE

29 CREATININE 2 x 100 µl PP tubes Photometry, Jaffé method

30 COTININE 2 x 5 ml PP tubes GC-MS

31 METALS 8 x 3 ml PP tubes ICP-MS

32 ALIQUOTING AND STORAGE - 20ºC until the analysis

33

34 Esteban M, Jimenez J, Schindler B, Holger K, Angerer J, Castaño A.