Journal of Cultural Heritage 13 (2012) 8997

Case study

Identication of natural dyes in historical textiles from Romanian collections by

LC-DAD and LC-MS (single stage and tandem MS)
Irina Petroviciu a,,e , Ina Vanden Berghe b , Ileana Cretu c , Florin Albu d , Andrei Medvedovici e
a

National Museum of Romanian History, 030026 Bucharest, Romania

Royal Institute for Cultural Heritage, 1000 Brussels, Belgium
c
National Museum of Art of Romania, 010063 Bucharest, Romania
d
Bioanalytical Laboratory, S.C. LaborMed Pharma S.A., 032266 Bucharest, Romania
e
Department of Analytical Chemistry, Faculty of Chemistry, University of Bucharest, 050663 Bucharest, Romania
b

a r t i c l e

i n f o

Article history:
Received 7 February 2011
Accepted 5 May 2011
Available online 14 June 2011
Keywords:
Natural dyes
Liquid chromatography
Diode Array Detection
Mass spectrometry (single stage MS and
tandem MS)
Romanian historic textiles (religious
embroideries and brocaded velvets)

a b s t r a c t
In this study, the dyes present in ve 17th- to 18th-century textiles from the National Museum of Art
of Romania, three religious embroideries and two brocaded velvets, are characterized and discussed,
together with earlier results on textiles from Romanian collections obtained by the same research group.
Dye analyses were performed using two methods: the well-established liquid chromatography-diode
array detection (LCDAD) and a recently developed liquid chromatography-mass spectrometry (LCMS)
analytical protocol. The examination of very small historical samples by both techniques allows a better
insight in the advantages and limitations of the two approaches to real analyses to be obtained. LCMS
data interpretation is based entirely on the results accumulated for dye standards. Electrospray ionization
(ESI) was used in the negative ion mode and an ion trap served as mass analyzer. Both single stage (MS)
and tandem (MS/MS) mass spectrometric approaches were considered. The dyes and natural sources
identied by both analytical techniques are discussed in the historical context of the textiles, with respect
to earlier results collected for similar Romanian objects. The study showed that the dye sources found in
the 17th- and 18th-century Romanian velvets and embroideries were produced using a wide variety of
dye sources, suggesting inuences from Europe as well as from Asia Minor. Dye sources imported from
New World have been also detected. The range of biological sources is in very good correspondence with
earlier results obtained from textiles in the Romanian Collections. LC-MS (single stage and tandem MS)
approaches have been demonstrated to be valuable tools for dye identication in small-scaled samples
from historical textile objects only if sufcient knowledge on the dyes and their biological sources is rst
accumulated within experiments performed on standard dyes and standard dyed bers.
2011 Elsevier Masson SAS. All rights reserved.

1. Introduction and research aims

Dyes obtained from naturally occurring biological sources have
been used in textile dyeing since antiquity. The identication of
their use in historic pieces may provide useful information about
where, when and how these objects were created and may also
contribute to their conservation.
Several studies have been dedicated to the identication of dyes
from biological sources in various types of textile preserved in
Western European collections in the last 50 years [16]. In contrast,
objects in Eastern European museums and monasteries including

Corresponding author.
E-mail addresses: petroviciu@yahoo.com, irinapetroviciu@mnir.ro
(I. Petroviciu), ina.vandenberghe@kikirpa.be (I. Vanden Berghe),
adileana@yahoo.com (I. Cretu), orin alburo@yahoo.com (F. Albu),
avmedved@yahoo.com (A. Medvedovici).
1296-2074/$ see front matter 2011 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.culher.2011.05.004

studies on dyes in textiles from Romanian collections have only

recently been considered as subjects for characterization of the
dyes present [711].
In the present work, dye analysis on three religious embroideries and two brocaded velvets, dating from the 17th to 18th
centuries, from the National Museum of Art of Romania is presented and discussed, together with earlier results on textiles from
Romanian collections obtained by the same research group. Data
resulting from the liquid chromatographydiode array detection
(LCDAD) method of analysis are compared with those produced
using a newly developed liquid chromatographymass spectrometry (LCMS) analytical protocol, based on the progressive use of
single stage (MS) and tandem (MS/MS) mass spectrometry. The
comparison of the resulting pattern of dye constituents obtained
through application of the two alternative techniques offers a
very good insight into the advantages and limitations of the two
approaches to real historical problems, where the reality of degradation of the textiles and the very limited sample sizes available

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I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

must be faced. This results in a mutual validation of the analytical protocols developed for the identication of dyes in cultural
heritage textiles.
The biological sources of dyes identied in the ve Romanian
textiles using both techniques are evaluated in terms of period,
provenance and technique, and compared with earlier results
obtained from similar objects [810].
2. Experimental
2.1. Historical samples
Fibers about 0.5 cm long were sampled from three religious
embroideries and two brocaded velvets dated to the 17th and 18th
centuries from the collection of the National Museum of Art, Romania. Sample withdrawal was possible due to the fact that all the
objects had recently passed through the textile restoration workshop for conservation.
2.2. References
For each analytical technique, dedicated libraries of references
were used. These databases were built up in the laboratories of
three of the research group partners and consist of UV-visible
(UVvis), single stage and tandem MS data for the dye components
discussed [12,13].
2.3. Sample preparation
Individual samples (about 0.5 mg) of dyed laboratory standard or historic bers were extracted with hydrochloric acid
(37%)/methanol/water (2:1:1, v/v/v) and prepared according to
the procedures described in detail in [12] for LCDAD and [13]
for LCMS analysis. For green-coloured samples, where the presence of indigoid dyes must be checked, an additional extraction
step was included. For this, after removing the coloured solution
resulting from hydrochloric acid extraction, 200 L dimethyl formamide (DMF) was added to the coloured ber and the mixture
was heated at 140 C for 10 minutes. The solution was then centrifuged at 12,000 rpm for 5 minutes and the supernatant liquid was
transferred to an injection vial.
2.4. Instrumentation
2.4.1. LC-DAD
Waters LCDAD equipment was used, data acquisition and
treatment being made by the Empower Pro 2002 software. Separation was achieved on a LiChrosorb RP-18 column, 125 mm L 4 mm
i.d. 5 m d.p. The mobile phase consisted of a mixture of methanol
(solvent A), methanol in water (1/9, v/v) (solvent B) and an aqueous
(5%, v/v) solution of phosphoric acid (solvent C). Gradient elution
was applied according to the prole given below, which includes
the re-equilibration step.
Time

Solvent A (%)

Solvent B (%)

Solvent C (%)

0
3
29
30
35

23
23
90
23
23

67
67
0
67
67

10
10
10
10
10

The ow rate was set at 1.2 mL/min. The volume injected was
20 L from which 5 L will pass through the column for analysis.
The other 15 l is sent to the waste. Detection was made within a
200800 nm wavelength range, with a spectral resolution of 1.2 nm.
Chromatograms were integrated systematically at 254 nm and also
at one or more other wavelengths at which the optimum response

of the dye constituent is observed. Results are presented as the

relative composition of dyes at the wavelength(s) of integration.
2.4.2. LC-MSD
LCMS and LCMS/MS experiments were achieved on a system constructed from Agilent Series 1100 modules. Detection
was made through a MS/MS ion trap detector using an electrospray ionisation (ESI) ion source, operated in negative ion mode.
The control of the chromatographic system and data acquisition
were achieved with the Agilent ChemStation software LC 3D version 10.02, incorporating the MSD trap control, version 5.2, from
Brucker Daltronics. Separation was achieved on a Zorbax C18 column, 150 mm L 4.6 mm i.d. 5 m d.p., thermostated at 40 C.
The mobile phase consisted of a mixture of aqueous 0.2% (v/v)
formic acid (solvent A) and methanol/acetonitrile (1:1 v/v, solvent B). Gradient elution was applied according to the prole given
below, which includes the re-equilibration step.
Time
0
5
10
16
18
18.01
22

Solvent B (%)
15
25
55
100
100
15
15

The ow rate was set at 0.8 mL/min. The injected volume was
5 L (from a total of about 200 L resulting from the sample
preparation). Several injections from the same solution may be performed, as described in the Results section below. The DAD detector
was placed in series between the column and the MS ion source.
UVvis spectra were acquired over the 200800 nm range with a
resolution of 2 nm. MS detection was made in the negative ion mode
with the following ESI operational parameters: drying gas temperature 350 C; drying gas ow rate 12 L/min; nebulising gas pressure
65 psi; capillary high voltage 2484 V. The ion trap used a maximum accumulation time of 300 ms and a total charge accumulation
(ICC) of 30000. The multiplier voltage was set at 2000 V and the
dynode potential at 7 kV. When working in the MS/MS mode, the
spectral width was 4 a.m.u. and the collisional induced dissociation
amplitude 1.6 V.
Automated Mass Spectral Deconvolution and Identication System (AMDIS) software was used as a complementary identication
tool. Chromatograms obtained with full scan single stage mass
spectrometric detection were exported in the Agilent MS Engine
format (.ms) and analyzed with the AMDIS software [13].
3. Results and discussions
Samples described in Table 1 were analyzed by LCDAD and
LCMS.
Fig. 1 shows the 17th-century Epimanikia (right hand sleeve,
after cleaning) described in Table 1.
Table 2 summarizes DAD and MS data obtained for the samples analyzed. For MS analysis, the detection of dyes was made
according to a dedicated analytical protocol, described in detail in
an earlier publication [13]. It includes chromatographic separation
and single MS full scan (FS) detection, followed by data processing through ion extraction chromatograms (IEC) according to m/z
values of the molecular ions of dyes in the database. Results are correlated with UVvis spectral data. For minor compounds the sample
was re-injected using single stage MS detection in the selected ion
monitoring/multiple ion monitoring modes (SIM/MID) as well as
by using tandem MS, product ion scan/single reaction monitoring
(SRM)/multiple reaction monitoring (MRM), for unambiguous conrmation of dye components. The conrmation procedure for the
dye constituents (single MS-FS, single MS-SIM or tandem MS) may

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91

Table 1
Characterization of samples discussed in the present study.
Textile object (period)

Function of the threads

Color

Sample code

Bedernita,
Religious embroidery (1746)

Lining
Sewing thread
Embroidery thread
Thread sewing the lining

Green
Pink yellow
Green
Kaki

A1
A12
A13
A14

Bedernita
(lining), religious embroidery
(1746)

Embroidery thread
Sewing thread
Sewing thread

Kaki
Green
Brown

B15
B17
B20

Epimanikia, Religious embroidery

(17th c.)

Embroidery support (edge)

Embroidery support (edge)
Embroidery support (edge)
Lining
Embroidery thread
Embroidery thread
Embroidery thread

Green
Pink
Pink yellow
Yellow
Orange
Red
Green

D23a
D23b
D23c
D24
D26
D27
D29

Sakkos,
Brocaded velvet
(17th c.)

Embroidery thread
Embroidery thread
Silk core metallic embroidery thread
Lining
Lining

Pink yellow
Red
Pink yellow
Green
Red

F37a
F37b
F38
F39
F40

Robe of Princess, brocaded velvet (1617th c.)

Weft

Green

E34

be directly correlated with the occurrence of the specic analyte in

the sample (Table 2).
When the semi-quantitative evaluation made using diode array
detection (indicated through normalisation of a peak area to the
sum of the peak areas in the chromatogram) gives a result for the
constituent of interest greater than 10%, the FS operating mode is
usually sufcient for the MS detection. If the semi-quantitative DAD
evaluation for the constituent is between 1 and 10%, single stage MS
in the SIM mode or MS/MS approaches are necessary for successful
MS detection. When DAD detection indicates the occurrence of a
compound at a level below 0.5%, it may be necessary to use AMDIS
deconvolution software for MS detection.
3.1. Red dyes
Red dyes were detected in eight samples, four from religious
embroideries and four from brocaded velvets. Anthraquinone dyes
were present in all the cases where samples still have a visibly
red color (3/8 samples). In one of these samples carminic acid,
the main dye component in Mexican cochineal (Dactylopius coccus Costa), Armenian cochineal (Porphyrophora hameli Brandt) and
Polish cochineal (Porphyrophora polonica L.), was detected by DAD.
Its presence was also conrmed by MS in both cases. Three minor

compounds, dcII (the C-glucoside of avokermesic acid [14,15]),

kermesic and avokermesic acids are also present in bers dyed
with cochineal. In the present work, only dcII was detected by the
presence of its molecular ion (m/z = 475), according to the FS-Ion
Extracted Chromatogram (IEC). The presence of kermesic and avokermesic acids was conrmed by the proles of their fragments
after spectral deconvolution with AMDIS (Fig. 2).
Based on the calculation between the ratio of the minor compounds and carminic acid in HPLC-DAD analysis [1618], the
biological source in F37b was established as Mexican cochineal
(D. coccus Costa).
For two other red samples alizarin and purpurin, the main
anthraquinone constituents in madder (Rubia tinctorum L.),
were detected by both DAD and MSD. The presence of minor
anthraquinone compounds from madder, anthragallol, munjistin,
xanthopurpurin and rubiadin was also established.
In ve cases described as pink or pink yellow a marker compound for redwood (Caesalpinia spp.) dyeings, called srwsoluble
redwood according to Wouters [4] or type C by Nowik [19] was
identied by both DAD and MSD. For the latter, identication was
made by FS-single stage MS followed by IEC of m/z = 243 a.m.u., the
major ion of srw according to the literature [13,20]. Conrmation
of this identication was achieved by detection by single stage MS
in the SIM mode and by MS/MS.

3.2. Yellow dyes

Fig. 1. Epimanikia described in Table 1. Religious embroidery (17th century) worked

in the Byzantine tradition, preserved in the Art Collection Museum, National
Museum of Art of Romania, inv. 88371.

Flavonoid yellow dyes were detected in 15 out of a total of 20

analyzed samples.
Dyers broom (Genista tinctoria L.) was identied as the main
biological source in ve samples and as a second biological source in
two other samples. In all the green samples blue indigo dyes (from
Isatis tinctoria L., Indigofera spp. or other species, discussed below)
were also present. The identication of dyers broom was based on
the detection of luteolin, genistein and apigenin, by both LCDAD
and MS detection techniques. In almost all cases, chrysoeriol a
minor compound recently identied in both weld and dyers broom
[13,15,21,22] was also detected by single stage MS in SIM mode
or by MS/MS.
Luteolin and apigenin without the presence of genistein were
identied by both DAD and MSD in ve samples, only one being
from brocaded velvet, a dyestuff constituent prole suggesting

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Table 2
Results obtained by alternative diode array and mass spectrometric (MS or MS/MS) detection modes.
Sample code Visual color

Results

Biological source
Dye constituents MSDb

Main source(s)

Traces

51 lu, 29 ge, 8 ap, 2 chry, 10 in

77 srw, 7 qu, 1 kf,
7 rht, 2 rhz, 6 ea
90 lu, 6 ap, 2 chry, 2 in
32 dat, 5 ge + lu, 4 kf, 1 irht, 1 ap, 23 in, 28 em, 5 chrys

lu(1), ge(1), ap(1), chry(2), in(2)

srw(1), qu(2), kf(2), ea(2)

Dyers broom and indigo

Redwood and buckthorn berries

Tannin plantc

lu(1), ap(1), in(2)

lu(1), ge(2), ap(2), dat(1,3), em(1), in(2)

Weld and indigod

Bastard hemp, rhubarbe and indigo

Dyers broom

lu(1), ap(2), dat(1,3), em(1), kf(2), in(2)

Bastard hemp, rhubarb, weld or eq. and indigo

lu(1), ge(1), ap(1), chry(2), in(2),

lu(2)
lu(1), ge(1), ap(2), chry(3), in(2)
srw(1), ca(1,3), ea(2), fk(*), ka(*)

Dyers broom and indigo

Weld (or another avone-containg plant)
Dyers broom and indigo
(Mexican) cochineal, redwood and tannin plant

A1
A 12

Green
Yellow pink

A 13
A 14
B 15

Green
Kaki
(yellow/green)
Kaki

B 17
B 20
D 23 a
D 23 b

Green
Brown
Green
Pink

D 23 c
D 24

Pink yellow
Yellow

D 26
D 27
D 29

Orange
Red
Green

F 37 a
F 37 b

Yellow pink
Red

F 38

Pink yellow

68 srw, 14 , 15 sul, 3 ea
71.5 ca, 1.5 dcII, 1
fk, 0.5 ka, 25.5 ea,
[288 nm: 0.8 dcII,
97.7 ca, 1.5 fk + ka]
81 srw, 10 , 9 sul, +ea

F 39
F 40
E 34

Green
Red
Green

39 lu, 46 ge, 12 ap, 1 chry, 2 in

87 al, 12 pu, +ag
57 lu, 5 ap, 1 chry, 37 in

18 dat, 25 lu,
1 ap, 19 in, 32 em,
1 kf, 4 chrys
43 lu, 43 ge, 10 ap, 1 chry, 3 in
92 lu, 8 ap
32 lu, 66 ge, 1 ap, +chry, 1 in
64.5 ca, 12 srw,
23 ea, +fk, 0.5 ka
90 srw, 1 lu, 9 ge
54 lu, 36 ge, 2 ap,
1 chry, 5 ea, 2 qu,
+, +sul
35 , 56 sul, 8 ea, + kf
69 al, +xp, 25 pu, 1 ru, +ag,
46 lu, 4 ap, 2 chry, 43 in, 4 ea, 0.5 , 0.5 sul

Redwood
srw(1), lu(2), ge(2), ap(3)
lu(1), ge(1), ap(2), chry(2), (3), sul(3), ea(2,3), qu(3), Dyers broom and redwood

Dyers broom
Young fustic and qu based dye

(1,3), sul(1,3), ea(2)

pu(1), ru(1), al(1), mu(1), xp(1), ag(2)
lu(1), ap(2),
chry(2),
(2), sul(2), in(2)
srw(1,2), (2), sul(2), ea(2)
ca(1), dcII(2), fk(*), ka(*), ea(2)

Young fustic
Madder
Weld and indigo

Young fustic

Redwood, young fustic

(Mexican) Cochineal and tannin plant

srw(1,3), (2),
sul(2),
ea(2)
lu(1),ge(1), ap(1), chry(2), in(2)
al(1), pu(1), ru(1), ag(2)
lu(1), ap(2), chry(2), in(2)

Redwood, young fustic

Dyers broom and indigo

Madder
Weld and indigo

The following abbreviations were used: al: alizarin; ag: anthragallol; ap: apigenin; ca: carminic acid; chrys: chrysophanic acid; chry: chrysoeriol; dat: datiscetin; ea: ellagic acid; em: emodin; : setin; fk: avokermesic acid
(also called laccaic acid D); dcII: avokermesic acid, C-glucoside; ge: genistein; in: indigotin; irht: isorhamnetin; kf: kaempferol; ka: kermesic acid; laA: laccaic acid A; lu: luteolin; mu: munjistin; pu: purpurin; qu: quercetin;
rht: rhamnetin; rhz: rhamnazin; ru: rubiadin; srw: soluble redwood; sul: sulfuretin; xp: xanthopurpurin.
a
Numbers before the abbreviation for a dye represent the relative composition (%) corresponding to peak areas integrated in the chromatogram monitored at 255 nm (unless specied in the column); + indicates values
lower than 0.5%.
b
Numbers between brackets have the following meaning: (1) detected through single stage MSFS/IEC; (2) detected through MSSIM/MID modes; (3) detected through MS/MS (MRM, product ion scan); (*) is used for dyes
evidenced through deconvolution by AMDIS software.
c
The term tannin plant is used as a shorter name for tannin-producing plant and is not indicative of a particular dye source.
d
Indigo refers to several plants that produce indigo.
e
The term rhubarb should be read as rhubarb or another emodin-containing plant.

I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

Dye constituents
DADa

I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

93

Fig. 2. LCMS and LCMS/MS of sample F37b (red), where (Mexican) cochineal was identied. Upper image, from top to bottom, the UVvis chromatogram; MSFS, IEC for
carminic acid (molecular ion m/z = 491 and ion produced by decarboxylation in the source m/z = 447); IEC for dcII (molecular ion m/z = 475 and ion produced by decarboxylation
in the source m/z = 431); IEC for ellagic acid. Lower images: detail of ion proles after AMDIS deconvolution for avokermesic (molecular ion m/z = 313 and ion produced by
decarboxylation in the source m/z = 269) and kermesic (molecular ion m/z = 329 and ion produced by decarboxylation in the source m/z = 285) acids. The gure illustrates the
exibility in use of mass spectrometry which allows the gradual detection of minor compounds.

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I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

Fig. 3. LCDAD chromatograms for sample B15, integrated at 255 and 350 nm, respectively, with detection of datiscetin, emodin, indigotin, luteolin, apigenin, kaempferol
and chrysophanic acid, suggesting the use of bastard hemp, weld (or another avone-containing plant; indigo/woad and rhubarb (or another emodin containing plant) the
compound marked with ? has similar UV spectrum to datiscetin, but different retention time.

the use of weld (Reseda luteola L.). However, recent studies have
demonstrated that other biological sources containing luteolin and
apigenin, of which sawwort (Serratula tinctoria L.) is the most
well known [14,15], also exist and could have been used as textile dyes. As a consequence, only those samples where chrysoeriol
was detected together with luteolin and apigenin are likely to
have been dyed using weld; in the other cases the use of another
avone-containing plant may not be excluded. When the presence of weld (or another avone-containing plant) corresponded
to green-coloured bers, indigo dyes were also detected, while
in another sample a combination of rhubarb (or another emodincontaining plant), bastard hemp and indigo dyes were found to
have been used.
Fisetin and sulphuretin, the main dye components in young fustic (Cotinus coggygria L.) were detected in three samples within
the present study. Young fustic was detected as a single biological source in one case and was identied together with soluble
redwood in pink-yellow samples. Traces of young fustic were also
detected in two additional samples where setin and sulphuretin
were detected by single stage MS in SIM mode or by MS/MS product
ion scan.
Datiscetin, the main dye component in bastard hemp
(Datisca cannabina L.) was identied by DAD in two kakicoloured samples, in both cases together mainly with emodin
and with chrysophanic acid, kaempferol and isorhamnetin as
minor constituents, as well as avonoids from either dyers
broom or weld (or another avone-containing plant) and
indigo (Fig. 3). Poor chromatographic resolution is obtained
between datiscetin and luteolin. Additionally, another compound exhibiting a very similar UVvis spectrum to datiscetin,
but having increased retention, could be observed in the
chromatogram.
Evidence for the presence of datiscetin (m/z = 285) in both samples was based on the MS information collected from a standard
sample of wool dyed with bastard hemp. As no other dye was
reported in this source, the identication of datiscetin by MS was
also conrmed by MS/MS analysis, based on the fragmentation pattern obtained through product ion scan. In LCMS separations, as

illustrated in Fig. 4, datiscetin co-elutes with kaempferol; however,

comparison of the MS/MS product ion scan spectra allowed identication of both datiscetin and kaempferol in the kaki-coloured
sample B15.
Rhamnetin was identied by DAD together with quercetin,
kaempferol and rhamnazin in an ochre-yellow sample (A12), suggesting the use of berries from a species of buckthorn (Rhamnus
spp.). Analyses of two wool references, dyed with either the bark
or the fruits (berries) from alder buckthorn (Rhamnus frangula L.)
resulted in the detection of mainly emodin and minor amounts
of rhamnetin and quercetin for the former, bark-dyedsample, and
mainly rhamnetin, isorhamnetin and quercetin for the latter. The
components detected in sample A12 by MS thus conrm the use of
berries from a Rhamnus species as the source of dye.
Emodin, the main dye component in alder buckthorn bark,
rhubarb, yellow dock and other biological sources was also detected
in two samples. It was not possible to evidence the presence of
chrysophanic acid (m/z = 253) by MS, based on the IEC, nor by the
identication of the fragment m/z = 239, which would correspond
to the fragment induced by de-methylation. A more detailed study
on a pure standard of chrysophanic acid (not available at the time
of the present experiments) is needed in order to be able to identify it in historical samples. The limited information on these two
anthraquinone dyes of vegetable origin, emodin and chrysophanic
acid, as textile dyes may be explained by their rare identication in
historical textiles. Like emodin, the presence of chrysophanic acid
may indicate the use of alder buckthorn bark (Rhamnus frangula
L.), or the roots of rhubarb (Rheum sp.) or dock (Rumex sp.) [2325].
However, no systematic study in which the biological source of the
dye has been identied unambiguously as one or other of these
plants has so far been reported.
3.3. Blue dyes
Although blue-coloured samples were not selected for analysis, indigotin was detected in eight green-coloured samples. For
mass spectrometric detection, FS-single stage MS in SIM mode
according to the molecular ion of indigotin (m/z = 261) was used.

I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

95

Fig. 4. LCMS and LCMS/MS data from sample B15, supporting the identication of datiscetin, emodin, luteolin, apigenin, and kaempferol (identication for indigotin is
not given; chrysophanic acid, although detected by LCDAD, was not detected by LCMS).

Indigotin is the main dye component in woad, Isatis tinctoria L.

and in the indigo plant itself, Indigofera spp. but no analytical
method has been reported to distinguish between these species
[26]. The term indigo is thus used to refer to indigotin-producing
plants.

3.4. Tannin
Ellagic acid was detected in seven samples with both detection techniques. The presence of ellagic acid indicates the use of
a tannin-containing plant material either for textile dyeing or for

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I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

Table 3
Biological sources attributed to samples analyzed throughout the present study, compared to other results for samples from religious embroideries and brocaded velvets
from Romanian collections previously identied through the LCDAD method by the same research group [810].
Religious embroideries
15th-16th c.

Cochineal
All
(DC)
Kermes
Lac dye
Madder
Safower
Redwood
Young fustic
Weld (or another avone-containing plant)
Dyers broom
Buckthorn berries
Rhubarb (or another emodin-containing plant)
Bastard hemp
Tannins
Logwood
Indigoid

6
(3)16th c.
3
21
29
2
24
24
26
8
0
2
1
39
0
40

Brocaded velvets

17th18th c.

19th c.

Present

Previous

1
(1)
0
0
1
0
3
1
4
4
1
2
2
1
0
5

3
(1)
0
0
3
1
4
1
3
0
3
2
2
5
1
7

weighting silk [4,24]. In the present study, a high proportion of

ellagic acid from a tannin-producing plant source was detected
twice in red samples in combination with cochineal while much
lower proportions were detected in a yellow pink sample together
with redwood and buckthorn berries. Ellagic acid was also detected
in all the ve cases when setin and sulphuretin were identied,
indicating the use of the dye from the heartwood of young fustic.
This is probably due to the presence of a trace of the tannins present
in young fustic (primarily in the leaves and twigs).
3.5. Discussion on the biological sources together with previous
results on religious embroideries and brocaded velvets from
Romanian collections
Attribution of the biological sources of dyes identied and conrmed in the analyzed samples is summarized in Table 2. All the
sources detected in the present series of analyses were also identied in one or more groups of samples analyzed before by the same
research team. According to the studies performed until now, six
sources of red dye have been identied in religious embroideries
and brocaded velvets from Romanian collections dating from the
15th to the 19th centuries (Table 3).
Half of these are of animal origin (cochineal, kermes and lac) and
half derive from plant sources, and from various parts of the plants:
roots (madder), petals (safower) and wood (redwood). Cochineal
(both Old and New World) and redwood were detected in all the
textile groups studied; lac dye, madder and safower were only
present in embroideries. The combination of lac dye and madder
was the main source of red used in the support fabric for embroideries in the 15th and 16th centuries. According to literature, lac
was hardly used in Europe for textile dyeing, but only for dyeing
leather and as an organic pigment. It was mentioned as imported
into the Ottoman Empire as early as the 15th century and, according to literature, it has been detected in Ottoman textiles [27,28]. Its
use in religious embroideries would thus suggest an Oriental origin
for these materials.
Kermes was only identied in religious embroideries and brocaded velvets dated to the 15th and 16th centuries, which is in
accordance with literature mentioning that, due to its lower cost
and ease of use Mexican cochineal eventually became the only animal source of red dye used in Europe, soon after the discovery of
the New World [25].

1
(0)
0
0
0
0
3
3
4
1
2
1
0
2
1
5

15th16th c.

2
(0)
1
0
0
0
4
3
7
0
0
0
0
6
0
3

17th c.
Present

Previous

1
(1)
0
0
1
0
2
2
1
1
0
0
0
0
0
2

0
0
0
0
0
0
0
1
0
2
0
0
0
3
0
1

Six sources of yellow avonoids were identied in the series,

weld (or another avone-containing plant), young fustic and dyers
broom being the most widely used. Except for the latter that was
not present in 15th16th century embroideries, the others were
detected in all the groups considered. Bastard hemp, buckthorn
berries and rhubarb (or another emodin-containing plant) were
only detected in embroideries, bastard hemp up to the 18th century, buckthorn berries not before the 17th century and rhubarb (or
another emodin-containing plant source) in textiles from the 15th
to the 18th century. However, both bastard hemp and rhubarb (or
another emodin-containing plant) were only detected in Ottoman
textiles [28], which also supports the suggestion of an Oriental
origin for the materials.
As far as the blue dyes are concerned, indigo was identied in
religious embroideries and brocaded velvets from the 15th to the
19th century, while logwood was only present in 17th- to 19thcentury embroideries. This is no surprise when we remember that
this source was only available after the discovery of the New World
[29].
4. Conclusions
All the dyes identied on the 17th- and 18th-century Romanian velvets and embroideries are in very good correlation with the
existing knowledge on dyes and biological sources used in Europe
and Asia Minor during this period [1,2,5,6,24,25,28]. The results
may be also correlated in terms of period and ber function with
previous data obtained for similar textiles by the same research
team [810]. From the whole group of analyses performed up till
the present on textiles from Romanian collections, it may be concluded that the combination of lac dye and madder was the main
source of red in the 15th- to 16th-century embroideries, while kermes was used when a more precious textile was intended. Mexican
cochineal was preferred in later textiles. Lac dye, bastard hemp and
rhubarb (or another emodin-containing plant) were only detected
in embroideries. Based on this fact and considering that these dye
sources were not identied in textiles from Europe, but only in
Ottoman pieces, it may be stated that at least part of the materials
used in embroideries dating from the 15th to the 18th centuries
have an Oriental origin.
As far as the techniques applied are concerned, it can be concluded that a good correlation between the results produced

I. Petroviciu et al. / Journal of Cultural Heritage 13 (2012) 8997

by LCDAD, LCMS and LCMS/MS was found, not only for the
major dye components, but also for the minor accompanying constituents. For some of the minor components co-eluting under the
chromatographic conditions of the LCMS method, spectral deconvolution with AMDIS software was established. The study showed
that LCMS and LCMS/MS approaches were conrmed as versatile tools for the identication and conrmation of dyes in historic
textiles, if consistent work is rst accumulated on a collection of
standard dyes and dyed bers for construction of suitable spectral
libraries.
Acknowledgements
The authors would like to thank the National Museum of Art of
Romania and the Putna Monastery, Romania for access to their collections. They are also grateful to LaborMed Pharma, who offered
an open access to the LCMS/MS analytical instrumentation and to
Ms Marie-Christine Maquoi from the KIK/IRPA laboratory for the
expert assistance in LCDAD dye analysis. Professor Recep Karadag
from Marmara University, Istanbul, Turkey, who offered a bastard
hemp-dyed bre, is also acknowledged. The authors are also grateful to Ms. Jo Kirby, National Gallery London Scientic Department
(retired), for carefully reading and improving the English text.
References
[1] J.H. Hofenk de Graaff, W.G. Th Roelofs, On the occurrence of red dyestuffs in
textile materials from the period 14501600, in: 4th Meeting ICOM Committee
for Conservation, Madrid, 1972.
[2] J.H. Hofenk de Graaff, T.W.G. Roelofs, The analysis of avonoids in natural yellow dyestuffs occuring in ancient textiles, in: 5th Meeting ICOM Committee for
Conservation, Zagreb, 1978, p. 115.
[3] J. Wouters, Analyse des colorants des tapisseries brugeoises, in: Bruges et la
tapisserie des xvie et xviie sicles, Mouscron, Bruges, 1987, p. 515526.
[4] J. Wouters, Dye analysis of Florentine borders of the 14th to 16th centuries,
Dyes in History and Archaeology 14 (1995) 4858.
[5] I. Karapanagiotis, L. Valianou, Y. Sister Daniila, Chryssoulakis, Organic dyes in
Byzantine and post-Byzantine icons from Chalkidiki (Greece), Journal of Cultural Heritage 8 (2007) 294298.
[6] M. Van Bommel, J.H. Hofenk de Graaff, Master dyers to the court of Sicily, Dyes
in History and Archaeology, 22/23, publication due 2012.
[7] I. Petroviciu, J. Wouters, Analysis of natural dyes from Romanian 19th and 20th
century ethnographical textiles by DAD-HPLC, Dyes in History and Archaeology
18 (2002) 5762.
[8] I. Petroviciu, J. Wouters, I. Vanden Berghe, I. Cretu, Dyes in some textiles from
the Romanian Medieval Art Gallery, Dyes in History and Archaeology 22/23,
publication due 2012.

97

[9] I. Petroviciu, J. Wouters, I. Vanden Berghe, I. Cretu, Dye analysis on some 15th
Century byzantine embroideries, Dyes in History and Archaeology. 22/23, publication due 2012.
[10] I. Petroviciu, I. Vanden Berghe, I. Cretu, J. Wouters, Analysis of Dyestuffs in
15th17th Century byzantine embroideries from Putna Monastery, Romania,
Dyes in History and Archaeology, 24/25, publication due 2012.
[11] M. Trojanowicz, J. Orska-Gawrys, I. Surowiec, B. Szostek, Chromatographic
investigation of dyes extracted from Coptic texiles from the National Museum
in Warsaw, Studies in Conservation 49 (2004) 115130.
[12] J. Wouters, N. Rosario-Chirinos, Dye analysis of pre-combian peruvian textiles
with HPLC and DAD, Journal of the American Institute for Conservation 31
(1992) 237255.
[13] I. Petroviciu, F. Albu, A. Medvedovici, LC/MS and LC/MS/MS based protocol
for identication of dyes in historic textiles, Microchemical Journal 95 (2010)
247254.
[14] D. Peggie, The development and application of analytical methods for the identication of dyes on historical textiles, PhD thesis, University of Edinburgh
(2006).
[15] D. Peggie, A. Hulme, Mc. Nab, H.A. Quye, Towards the identication of characteristic minor components from textiles dyed with weld (Reseda luteola L.) and
those dyed with Mexican cochineal (Dactylopius coccus Costa), Microchemica
Acta 162 (2008) 371380.
[16] J. Wouters, A. Verhecken, The Coccid insect dyes: HPLC and computerized analysis of dyed yarns, Studies in Conservation 34 (1989) 189200.
[17] J. Wouters, A. Verhecken, The scale insect dyes (Homoptera:Coccoidea) species
recognition by HPLC and Diode-Array analysis of the dyestuffs, Annales de la
Socit entomologique de France 25 (1989) 393410.
[18] I. Vanden Berghe, Investigation of cochineal dyeing parameters and renement
of the cochineal identication system, Dyes in History and Archaeology22/23,
publication due 2012.
[19] W. Nowik, The possibility of differentiation and identication of red and blue
soluble dyewoods: determination of species used in dyeing and chemistry of
their dyestuffs, Dyes in History and Archaeology 16/17 (2001) 129144.
[20] I. Karapanagiotis, E. Minopoulou, L. Valianou, Y. Sister Daniilia, Chryssoulakis,
Investigation of the colourants used in icons of the Cretan school of iconography, Analytica Chimica Acta 647 (2009) 231242.
[21] E. Ferreira, New approaches towards the identication of yellow dyes in 463
ancient textiles, PhD thesis, University of Edinburgh (2001).
[22] http://www.organic-colorants.org.
[23] H. Schweppe, Handbuch der Naturfarbstoffe, Vorkommen, Verwendung, Nachweis, Nikol Verlagsgesellschaft mbH&Co, KG, Hamburg, 1993, p. 224228.
[24] J.H. Hofenk de Graaff, The colourful past. Origins chemistry and identication
of natural dyestuffs, Abegg Stiftung & Archetype Publications Ltd, Riggisberg
and London, 2004.
[25] D. Cardon, Natural dyessources, tradition, technology science, Archetype Publications, London, 2007.
[26] J. Wouters, Possible future developments in the analysis of organic dyes, Dyes
in History and Archaeology 20 (2005) 2329.
[27] I. Vanden Berghe, M.C. Maquoi, J. Wouters, Dye analysis of Ottoman silk, in: M.
van Raemdonck (Ed.), The Ottoman silk textiles from the Royal Museums of Art
and History in Brussels, Turhout Brepols, Brussels, 2004, pp. 4960.
[28] N. Enez, H. Bhmer, Ottoman textiles: dye analysis, results and interpretation,
Dyes in History and Archaeology 14 (1995) 3944.
[29] F. Brunello, The art of dyeing in the History of Mankind, Vicenza, 1973,
p. 175221.