HPLC and SEM-EDX analysis Results of HPLC and SEM-EDX analysis - Dyed textiles - Dye bath - Organic pigments Maarten van Bommel

Report of HPLC analysis (ICN) ICN report usually exist of: - Summary - Introduction, aim of the research - General description of the work done - Results and discussion - Conclusion - References and additional information - Analytical reports - Microscopy - FTIR - XRF - GC-MS - HPLC-PDA - SEM-EDX

Report of HPLC analysis (ICN) Analytical report (HPLC): - Information about the sample and object - Short description of the procedure used - Graph at wavelength were most components of interest shows VIS absorption - Spectra of the components found - Discussion of the results

Dyestuff analysis of dye baths Object Weld (from Anjou) dyebath samples, before, after 1st and 2nd bath Sample preparation Directly from the baths Sample location Equipment HPLC with autosampler, PDA en fluorescence detection, 1 ul injection Request Dyestuff analysis Column Luna C18, 150x2 mm id. 3 m with Guard column. Column in oven 35°C. Date sample 30-03-2005 Gradient Buffer A: 10% methanol/90% water (v/v), Buffer B: 100% methanol, Buffer C: 5% phosphoric acid in water (w/v). Start gradient: 74%A/16%B/10%C. After injection in 15 min. to 0A/90B/10C, 5 min at that condition, next in 3 min to 0A/100B/0C, 4 min at that condition, next in 5 min. back to start condition. Sample taken by Maarten van Bommel Analysis done by Marjolein Groot Wassink & Maarten van Bommel Sample number Work number Object number Detection PDA, 200-700 nm, 1 scan/s, 1,2 nm resolution. Option: fluorescence detection at ex=370 nm and em=525 nm, with post-column addition of 0,1 M AlNO3 in 20%methanol/80%water at 0,2 ml/min. Doc. Map Artech-dye project Filename HPLC Artech-dye baths (weld) for wool and silk Date analysis 4-04-2005

HPLC-PDA Chromatogram 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 16.683 Absoption at 350nm (AU) 15.469 20.812 17.985 22.069 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00 Retention time (min)

UV-VIS spectra Luteolin-3’,7- diglucoside Apigenin glucoside? Luteolin

Results and Discussion Description of the components identified Explanation which dye source is identified Explanation about unknown components Explanation about un-assigned peaks Conclusion -Usually a short description of the dye source identified

HPLC-PDA Chromatogram 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 20.906 22.165 Absoption at 350nm (AU) 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00 Retention time (min)

HPLC-PDA Chromatogram

Results Textile samples -Luteolin and Apigenin found (99,5%) Other flavonoids at low concentration Ratio luteolin and apigenin calculated at 350 nm Just to compare the samples Wool, weld and potash (quadruple) Ratio luteolin:apigenin 90:10 Rsd area luteolin and apigenin 9% Rsd ratio luteolin:apigenin 2% Wool, weld only (duplo) 10% more dyestuff Ratio luteolin:apigenin 88:12

Results Textile samples Silk, weld and potash -Amount dyestuff compare to wool 30% -Ratio luteolin:apigenin 95:5 Silk, weld only -Amount dyestuff compared to wool the same -Ratio luteolin:apigenin 86:13

Results Textile samples Wool was also extracted with formic acid -Only 35% dyestuff extracted (compared to HCL) -Luteolin is main component -Luteolin 3’,7-diglucoside found -Luteolin 7 glucodide found Wool was also extracted with EDTA -Only 25% dyestuff extracted (compared to HCL)

Results pigment samples -Luteolin and Apigenin main components -Two other flavonoids were found as well (unidentified) -Amount organic colorant in pigment pigment 1>pigment3.2>pigment2>pigment 3

Results pigment samples Ratio Luteolin: apigenin: unknowns P1 87% 11% 2% P2 67% 10% 23% P3.1 76% 19% 4% P3.2 84% 12% 4%

Results SEM-EDX Elements found in wool -Sulphur (from the protein) -Traces of aluminium (mordant) -Traces of calcium (also in untreated wool) Elements found in silk -No mordant was found! Elements found in the pigments P1 – not analysed P2 – Calcium and trace aluminium P3 – Aluminium and trace of calcium