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

2. 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

3. 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

4. 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
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, 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

5. 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)

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

7. 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

8. 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)

9. HPLC-PDA Chromatogram

10. 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

11. 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

12. 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)

13. 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

14. 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%

15. 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