Results ofHPLC and SEM-EDX analysis

Maarten van Bommel

- Dyed textiles- Dye bath- Organic pigments

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 ChromatogramA

bsop

tion

at 3

50nm

(A

U)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

Retention time (min)5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00

15.4

6916

.683

17.9

85 20.8

1222

.069

UV-VIS spectra200.00

400.00 600.00

205.9

268.4

342.3

15.47

200.00

400.00 600.00

204.7

254.2349.4

16.68

200.00

400.00 600.00

267.2338.7

501.4524.6563.6629.6648.0694.6

17.98

200.00

400.00 600.00

207.0

254.2

350.6

551.4

20.82

200.00

400.00 600.00

208.2

267.2341.1

435.9479.5508.7538.0553.8602.7623.5651.7666.4694.6

22.07

Luteolin-3’,7-diglucoside

Luteolin-7-glucoside

Apigeninglucoside?

Luteolin Apigenin

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 ChromatogramA

bsop

tion

at 3

50nm

(A

U)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

Retention time (min)5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00

20

.90

6

22

.16

5

HPLC-PDA Chromatogramab

sorp

tion

at 3

50 n

m (A

U)

0.000

0.001

0.002

0.003

0.004

0.005

0.006

Retention time (min)14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00

12.9

44

18.2

17

19.7

68

20.9

06

21.6

72 22.1

6522

.745 24

.445

26.1

12

32.0

53

33.2

31

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 90:10Rsd 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)-Luteolin is main component-Luteolin 3’,7-diglucoside found-Luteolin 7 glucodide found

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: unknownsP1 87% 11% 2%P2 67% 10% 23%P3.1 76% 19% 4%P3.2 84% 12% 4%

Results SEM-EDXElements 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 pigmentsP1 – not analysedP2 – Calcium and trace aluminiumP3 – Aluminium and trace of calcium