Summary of Analytical protocols used for Dye and organic pigment analysis Maarten van Bommel

Analytical protocols Evaluation of techniques used - Not standardization of the techniques - Determine pro’s and con’s of the different techniques - Establish a standard format of how to describe an analytical technique - So that others can reproduce it All partners submitted the analytical protocols in the format they are used to

Analytical protocols Non-destructive: - X-ray fluorescence (XRF) - Fibre optic mid-FTIR - Fibre optic micro-RamanUNI-PG - UV-VIS fluorescence - UV-VIS colorimetry - 3D fluorescenceKIK / IRPA - SEM-EDX (samples are coated)ICN Not comparable equipment, except fluorescence

Analytical protocols Destructive: - HPLC-PDA (6 protocols) OADC KIK-IRPA NGL ICN GCI NMS/UoE - HPLC-MS(2 protocols) GCI(combined with PDA) NMS/UoE - HPLC-Fluorescence ICN(combined with PDA)

Sample pre-treatment Microscopic examination KIK / IRPA NGL ICN Weight samples OADC KIK / IRPA NMS / UoE ICN(not paint samples) Colorant extraction Methanolic HCl all partners except NGL Methanolic BF 3 NGL

Sample pre-treatment Evaporation Nitrogen flow OADC (60 °C) ICN Vacuum desiccators KIK / IRPA GCI Rotary evaporator NMS / UoE (40 °C) None NGL Dissolve Methanol / water KIK / IRPA GCI NMS / UoE Di-methyl formamide OADC ICN Add methanol NGL

Sample pre-treatment Removal of particles / precipitation Filtering prior evaporation GCI KIK / IRPA Centrifugation after evaporation and dissolution ICN

HPLC analysis All partners uses RP-HPLC with silica C18 based materials However, all partners uses different column materials and columns with different dimensions (length and id). As a results, flow rate varies from 0.01 ml/min to 1.20 ml/min Injection volumes varies as well Columns are thermo stated at 25 – 40 °C (except KIK / IRPA)

HPLC analysis HPLC solvent compositionRuntime (min) H2O / ACN / TFA OADC 35 NGL*240 H2O / MeOH/ phosphoric acid KIK / IRPA 35 NMS / UoE 35 ICN 50 H2O / MeOH / formic acid GCI 45 H2O / MeOH NMS / UoE (HPLC-MS) 40 Accuracy of solvent preparation? (ICN and NMS / UoE)

Photo diode array detection (PDA) Range Start191, 200, 210 or 250 nm End700, 750 or 800 nm Resolution Varies from 1.0 to 2.5 nm Specific wavelengths UV 250 nm, 254 nm, 255 nm, 270 nm, 275 nm, 288 nm, 290 nm VIS 330 nm, 350 nm, 370 nm, 420 nm, 435 nm, 491 nm 495 nm, 500 nm, 530 nm, 540 nm and 610 nm Note: OADC, GCI and NMS focus only on UV

Mass spectrometry (MS) Only GCI and NMS / UoE Negative ESI Nitrogen sheath gas No sheath flow Interface200 °C NMS / UoE 350 °C GCI Scanning50 – 1000 m/zGCI ?NMS / UoE

Conclusions Everybody uses different systems, question remains: - How are the results affected by the different parameters? - Can we establish a standard format for description of protocols for (HPLC) analysis And do we want that?

Proposed format - Name of protocol - Aim of protocol - Author(s) - Date of latest version or revision - Chemicals - reagent, grade, concentration, supplier, product number

- Materials - Vials - Columns (and pre-columns) - Filters - Glassware - Pipettes - etc - Equipment - Pump - Degasser - Autosampler / injector - Column oven - Detector Proposed format, continue Type Specification Supplier Product number ?

- Solvent preparation - Pre-examination - Sample pre-treatment - Analysis - Blank and standards - Detection parameters - Data evaluation Proposed format, continue