Title

Analysis of Fibre Dyes by High Performance Liquid Chromatography

Objective
To determine the fiber dyes in Pink, Brown and Red textile Sample

Introduction
High performance liquid chromatography (HPLC) systems are available for the
analysis of basic, acid and disperse dyes. HPLC systems have been developed in
this laboratory for the analysis of the dyes extracted from fibres. Casework size
fibres can be analysed by HPLC and batch type differences have been detected.
The addition of an autoinjector to the HPLC system is required to cope with the
number of fibres which are to be analysed in a typical case. The extraction of
dyes from fibres is usually carried out in capillary tubes, as for thin layer
chromatography (TLC). The fibre is placed in a capillary tube, covered with
extracting solvent, and the tube sealed and heated. The extract is removed using
a gas chromatography (gc) syringe, and diluted with the HPLC eluent for injection
onto the HPLC column. The extraction process is time consuming and some of the
extract can be lost with resultant reduction in peak size.
Dyes have been used to alter the appearance of cloth and fabric by changing
their colour since ancient times.(Garfield, 2000) Dyes can be defined as any
substance which has an affinity for a substrate and, on binding to that substrate,
alters the way in which the material absorbs light, and hence changes the colour
that is subsequently observed. Colour is due to the preferential absorption of
certain wavelengths of light, the observed colour being those wavelengths of
light that are reflected. The particular bands of light absorbed, and hence the
colour observed, by a particular dyed fabric is due to the chemical structure of
the dye. For this experiments, we used Alizarin, Carminic Acid and Martious
Yellow dyes.

Reagents and solutions

1.
2.
3.
4.

Methanol (HPLC Grade)

Hydrochloric Acid (37%)
Phosphoric Acid (5%)
References dyes solutions (100 ppm) : Alizarin, Carminic Acid, Martious
Yellow.

Apparatus
1.
2.
3.
4.
5.

Small beakers
Hot plate
Dessicator
Centrifuge
Sample tubes

Sample
3 coloures textile pieces.

Instruments
High Performance Liquid Chromatography (Agilent G1314A) equipped with
photoiodide array detector, 20 L sample loop and 5m RP C18 column.

Analytical Procedure
1. Sample Extraction
a) About 4 mg of textile fibre in 1.5 mL of H2O : methanol : 37 % HCI (1:1:2
v/v/v) was treated for 10 min at 100 oC in a small beaker.
b) The extract was cooled down and filtered by centrifuginal forces.
c) The filtrate was dried in desiccator over NaOH and in vacuum.
d) The residue was redissolved in an appropriate volume of methanol/water
(1/1, v/v).
2. Instrument Set-up

Wavelength : 200 to 320 nm

Mobile phase : (A) water, (B) methanol, (C) 5 % (w/v) phosphoric acid in
water.
Elution program : 66A:24B:10C for 2 mins, linear gradient to 0A:90B:10C
for 27 minbs, and hold on 0A:90B:10C for 3 mins.
Flow rate : 1.2 mL/min

3. Dye Analysis.
100 L of the sample extract was transfered into a clean, dried sample tube and
1 mL of the mobile phase was added and well shaked. 10 L of the diluted
sample was injected.

4. Identification of dye components in fabric extracts.

10 L of each references solutions individually injected to identify the main dye
components in the fabric extract.
Results
Standard
Martious Yellow
Alizarin
Carminic Acid

Retention Time (min)

5.760
5.312
2.805

Area
4473762
5612695
2669832

Table 1 : Retention Time of Standard Injected

Sample
Pink
Brown
Red

Retention Time (min)

5.365
2.325
5.845

Area
14251580
12459044
38415

Table 2 : Retention Time for Sample Injected

Discussion
In this experiment, we used high performance liquid chromatography to analyze
the extraction of textile fibers using photoiodide array detector to monitor the
absorbance at several wavelength. Based on the results that we obtained, for
pink textile sample, it contained Alizarin dyes because of the peak detected at
retention time for the pink (5.365 min) sample was closest to the retention time
only for the alizarin dyes (5.312 min) when we comparing both species. For
Brown textile sample, it contained Carminic Acid dyes because the closest of
peak detected at retention time for the brown sample (2.325 min) to the Carminic
Acid dyes (2.805 min). For the third sample, Red textile sample shows that it
contained Martious Yellow dyes due to the the closest peak detected at retention
time for the brown sample (5.845 min) to the Martious Yellow Dyes (5.760 min).
From the chromatograms that obtained, we found that, there are band
broadening at the peaks of martious yellow and pink textile sample. The band
was broaded due to the B/v effects. B/v effects causes the diffusion is 100 x less
in liquids than in the gas and can be neglected. Another factor that contribute to
the band broadening is Cv effects. For Cv effects, we assumed that mass
transport effects are the largest contribution. The mobile phase mass transfer
coefficient: CMv = (fM(k)dp2/DM)v. Smaller dp (particle diameter of packing)
increases the surface area/volume ratio and thus decreases mass transport in the
mobile phases.

Conclusion
We can coclude that the Pink textile sample contained Alizarin dyes, Brown textile
sample contained Carminic Acid dyes and the Red textile sample contained
Martious Yellow dyes.

References

RME GRIFFIN and SJ SPEERS (1995). Use of tapered vials for in situ extraction of
fibre dyes for
analysis by HPLC. Northern Ireland Forensic Science Laboratory, 151 Belfast
Road, Carrickfergus,
Northern Ireland, BT38 8PL. Retrieved November 13, 2012, from
http://www.sciencedirect.com.ezaccess.library.uitm.edu.my/science/article/pii/S13
55030695726624

http://espace.library.curtin.edu.au/webclient/StreamGate?
folder_id=0&dvs=1352791451625~57

Experiment 9:
Analysis of Fibre Dyes By High Performance
Liquid CHromatography
Name : Nur Zuraihan Binti Abd Wahab (2011856974)
Normahani Binti Ramli (2011603842)

Group : ASB3Ag

Name of Lecturer : Miss Anis Shuhada