Organic pigments analyses

Sanyova Jana
jana.sanyova@kikirpa.be

“Back to the Roots”

Workshop on the Preparation of Historical Lake Pigments
March 23th - 25th, 2011
Doerner Institut , München
 Plan

• Definition of organic pigment

• History of the analysis
• Diagnostic
• Analyses
– Substrate
– Dyestuffs
• Interpretation of dyes analysis
• Case studies
 Terminology
• Pigment
 The solid particles finely dispersed in the paint layer
organic pigment
 Pigment containing an organic dyestuff obtained
from a natural source (animal or vegetale) or by
synthesis. The dyes can be fixed or not on an inert
substrate

lake pigment
 Organic obtained by
complexation, precipitation or
pigment
chemisorption of the dyestuffs on
inert substrate, with low refractive
index, in general inorganic.
Preparation of a madder lake

1/ Extraction
extraction of water-soluble glycosides
and their deglycosylation into the aglycones

2/ Complexation
complexation of aglycones with
a metal cation (Al3+ S)

3/ Fixation onto the substrate

precipitation and/or addition of inorganic
substrate (alumina, gypsum, chalk, ) on which
the Al-dyestuff complexes adsorb

4/ Finalisation
filtration of the lake, rinsing of water-soluble
salts, drying and grinding
 GLYCOSIDES

O OH
O
O
O O CH2
HO OH
HO OH
OH
OH

-D-glucose
primverose
-D-xylose
O
Ruberythric acid = alizarin-2-primverosid
 Influence of parameters
bath temperature
précurseur

d'alizarine précurseur
lucidine

90 °C 20°C

20 °C 60°C
90°C
60 °C Inc à 27'
pseudopurpurine
précurseurs de alizarine
pseudopurpurine

purpurine
anthragallol
 Influence of parameters
color: countercation
, 040827
13:48

K Ca Na NH4
RTPARAM2.opj, GrBases,
Fri
alizarin
e
Na2CO3
(RT10)
2.5
pseudopurpurine
K2CO3
2.0
(RT2)
AUs*10/mg

Dyes recovery:
1.5
NH4OH (RT15)inc. à 27'
Na > K > NH4 > OH > Ca
1.0
précurseur CaCO3
d'alizarine précurseur
(RT9)
0.5 alizarin
de anthragallol
lucidine e

0.0

10 15 20 25

temps
[min]
 Chemical structure
Alizarin lake on amorphous alumina :
It is a chelate-type coordination compound, in which an Al(III) ion, at the
surface of amorphous alumina, forms a complex with one or two alizarin
molecules.
m = 576
+H z=
+3H2 0
O
n
In special circumstances,
i.e. at Al/aliz ratio so low
that no alumina can be
formed from the excess
aluminium, Al-aliz
complexes can polymerize
into linear chains, and then
slowly form gels by cross-
linking of these chains
through - interactions
between the aromatic rings
Sanyova. J. (2001), « Contribution à l’étude de la structure et des propriétés des laques de garance », PhD Thesis, l’Université Libre de Bruxelles
Analyses of lakes in the past
 Analyses in the past
Synthesis of alum
1806 Applied chemistry in
Arts

1807 Art de la peinture du

coton en rouge.

1809several communications in the Annales de Chimie

concerning the analyses of lake pigments found during the
excavation in Pompei

pinkish lake:
examined by smelling the products of calcination
=> lake of vegetal origin
Jean-Antoine
Jean-Antoine Chaptal (1809): Sur quelques couleurs trouvées à Pompeia, Annales de
Chaptal
(1756-1832) chimie et de physique, 70, pp. 22-31

Jo Kirby and Raymond White (1996) : The Identification of Red Lake Pigment Dyestuffs and the Discussion of their Use.
Technical Bulletin of National Gallery of London, Volume 17, pp. 56-80.
 Analyses in the past
1812 - Naples – rose pigment found in a broken vessel
during excavations
the Queen of Naples sent a sample to Davy

1815 – (23 février) - Davy presented his analytical results:

« it is a lake
pigment »
because:

- the sample was discoulored at the

surface of particles, but pink inside
Sir Humphry - its color is similar to the carmin
- the lost of weight by heating
Davy represented around 3 %
- it changed colour with pH
- the inorganic part of the sample is composed of
(1778-1829) silicates, aluminates and calcium carbonate

Jo Kirby and Raymond White (1996) : The Identification of Red Lake Pigment Dyestuffs and the Discussion of their Use.
Technical Bulletin of National Gallery of London, Volume 17, pp. 56-80.
 Analyses in the past
The pink pigment from Corinthe was identified
W.T. Russell by comparison of the reflexion curves of the
sample with that of a rose madder lake. These
en 1892 curves were obtained using a Beckman
spectrophotometer and are historically the first
instrumental analysis of the colour of lakes.

MARIE FARNSWORTH (1951): Second Century B. C. Rose Madder from Corinth and
Athens. American Journal of Archaeology, Vol. 55, No. 3 (Jul., 1951), pp. 236-239
Diagnostic of organic pigments
Colour + refractive index + UV fluorescence
Examination :
- macroscopic
- microscopic
Macroscopic examination
 Microscopic examination
Salt grain
needle

sample

Vis UV
ColourOpticka mikroskopia
+ refractive indexv+UV a Vis osvetleni
luminescence

Sanyova J., Saverwyns S. (2006) « Quelle technique dans

l’atelier de Lambert Lombard ? » Scientia Artis 3, p.259-295
Analyses of substrates
1/ atomic characterisation : SEM-EDX, XRF

2/ molecular characterisation: FTIR, MRS

 Staining with iodin

Confirmation by MRS

flour

Lead white
Wheat starch
 MRS in situ

 Renishaw plc

resolution 70µm
 MRS on cross-section

Advantages
 Analysis in reflexion mode =>x-section + in situ
 Excellent spectral & spatial resolution :
> 1 µm => 1 grain of pigment –
« finger-print » spectrum Drawback
 Identification of inorganic compounds  Fluorescence of
(substrates) and organics (mainly modern + OP
indigo)  Measurements
 Signal weak
long for OP
 SEM EDX on cross-section

C 54.117

Fragment « Couronnement
d’épine » Opitter (+/- 1540)
C 49.106

atomic and spatial information on substrate

 Master of Trebon,
NG Prague,
µ-FTIR in TR mode
1380

1233.03
1318.13

779.53
2853.57

798.38
3398.59

3076.37

2928.64

1650.88

1546.76
1453.56
1408.68

1112.58

879.46

721.15
670.49
612.17
1650
amideI
0.35

OH 1456
0.30

amideII
Al-O
0.10 0.15 0.20 0.25

S-O
Absorbance Units

oxalates
1318, 779
0.05

S-O
0.00

3500 300 2500 2000 150 1000

0 Wavenumber cm-1 0

Proteins-containing substrate => lake from wool shearings waste

 ToF – SIMS

Institut de Chimie des

Substances Naturelles

Saccharide
fragment

SANYOVA J., CERSOY S., RICHARDIN P., LAPREVOTE O., WALTER P., BRUNELLE A. Unexpected Materials in a Rembrandt
Painting Characterized by High Spatial Resolution Cluster-TOF-SIMS Imaging, Analytical Chemistry 2011, 83(3), pp. 753-760
 Dyestuffs analysis
1/ non-invasive spectroscopic methods (UV-Vis imaging, MRS, SIMS)

2/ chromatographic methods (HPLC,LC-MS, GCMS, TLC)

 MRS - indigo
Reliquaire de Sainte Ursule
(Bruges, 1400 – 1415)

SEM-EDX
X X

Indigo
Intensiteit / AU

Intensiteit / AU

Prussian blue

Lead white Lead white

500 1000 1500 2000

500 1000 1500 2000 2500

Ramans hift / cm-1

Ram ans hift / cm -1
 « Vis-Imaging »
Reflection µ-spectroscopy -– localisation and diagnostic

« VIS-imaging data » - non invasive (in situ) & non destructive (cross-section)
- digital treatement of ponctual spectras

spatiale resolution : ca 1µm => grains of pigment ( good for the mixtures)

spectrale resolution : 4 nm => in certain cases - possible identification 10 05_050126_03 47,25 min.spc - 1407.25 Min /
Bk

Indigo
tin
Jan Lievens : Vijf Muzen (1649-1652)

mAU
mAU
5 5

0 0

200 400 600

nm

indigo

b
OM imaging (at 600nm)

J. van der Weerd, and all : Identification of Pigments in Paint Cross Sections by
Reflection Visible Light Imaging Microspectroscopy, Anal. Chem. 2003, 75, 716-722
 « Vis-Imaging »
Vis µ-spectroscopy imaging : localisation & diagnostic

Patrick Caulfield:
Interior with a Picture
(1985-6)

c. 1: PR170, (a diazotized p-aminobenzamide coupled with 3-hydroxy-2-naphtho-O-phenetide)

c. 4: azo pigment – unidentified
c. 3,5,6,7: inorganic pigments : PbCl2, CdS, Fe2O3, TiO2
LC-PDA; LC-PDA-MS; MS
interface

Guard-

analytical column Column e

Vanne Ion trap
Vann
P
D
A
Detecteur MS
D
e
t
MS detector (LCQ Deca with ion trap MS detector)e:
c
Detector connected to the HPLC in addition tto the PDA UV detector
(but can also be used as independent mass spectrometry
e detector, for direct
injection of liquid samples) u
r
As HPLC detector, Mass Spectrometry outperforms classical detectors
(UV-VIS, conductivity, fluorescence, etc) in terms of selectivity
U (identification
power) and often of sensitivity V

6
0
0
0
 HPLC-PDA: Chromatograms & spectra
1: pseudopurpurin
purpurin
3 060919_09 30,26min.spc - 30.27 Min / Bkgd

750 750

1000 500 500

pseudopurpurin
1 250 250

0 0

2
alizarin 200 400
nm
600

2: alizarin
500
Abs [mAU]

060919_09 37,35min.spc - 37.35 Min / Bkgd

600 600

400 400

255 nm 200 200

495 nm (x 3) 0 0

200 600

0 400 nm

3: purpurin
060919_09 43,08min.spc - 43.08 Min / Bkgd
1000 1000

500 500

20 30 40 50 0 0

Time [min] 200 400

nm
600
 GC-MS
Neo-classical frame (1767–1800)
shellac (Sh),
sandarac
(S) mastic
(M). Dragonsblood
(Dae. Micracantha)

16th century reverse-glass

larch turpentine (L)

gum benzoin (B)
Dragon's blood
(Dracaena draco)

Ursula Baumer & Patrick Dietemann Identification and differentiation of dragon's blood in works of art using gas
chromatography/mass spectrometry, Anal Bioanal Chem (2010) 397:1363–1376
 From TLC to UTLC
TLC: thin-layer chromatography

Masschelein-Kleiner L; Microanalysis of hydroxyquinones in red lakes, Microchimica Acta, 1967, Vol.55, N° 6, Pages 1080-1085

HPTLC : high-performance thin-layer chromatography

smaller adsorbent material particle sizes and a slight reduction in layer thickness
UTLC: ultra-thin-layer chromatography
- no based on granular adsorbents, but have a monolithic structure based
on a silica gel matrix,
- no binder is needed to fix the layer on the glass
- ultra-thin layers has meso- and macro pores, with fine capillaries
penetrating
- increasing sensitivity => much smaller amounts of sample
- shortening analysis,
- reducing the quantity of consumables per analysis
M. Krauze-Baranowska, I. Malinowska, D. Głód, M. Majdan, A. Wilczańska; UTLC of Flavonols in Sambucus nigra
Flowers, JPC - Journal of Planar Chromatography - Modern TLC, Volume 22, Number 5, October 2009, p. 385-387
 Interpretation
What will influence the interpretation of dyestuffs analysis?
- sampling
- analytical protocol
- complementary analysis
- historical source study
 Sampling

Virgin’s Tunique – 11 red layers

Pieta 1400, Bratislava (Sk), Franciscain church
 Reconstitutions
• To understand the ancient recipes (influence of the
parameters)

• To search the link between the past and the

present

• To study the properties of organic pigments

• To understand the degradation mechanisms

• To look for « bio-markers »

• To develop new analytical methods

 Reconstitutions
précurseur Parameters
d'alizarine précurseur
lucidine
8

90 °C 20°C
6
20 °C 60°C
90°C
OD / mg

4
60 °C RT27
pseudopurpurine
précurseurs de alizarine
pseudopurpurine
2

purpurine
anthragallol

5 10 15 20 25
temps
Sanyova J. (2000), Étude des pigments organiques préparés à partir des racines des rubiacées européennes,
Art et chimie. La couleur, Actes du congrès, Paris, pp. 14 - 17.
 Study o the prepared samples

pH >0 0.2 1.5 1.6 1.8 1.9 2.1 2.5

HF
+Li
+Li/DTPA
+DFOM
HCl
Li
+DFOM
TFA Extraction of
Li alizarine at
+DFOM various pH
 Analytical protocol
HF
2N HCl
DMF/ MeCN 6M
2. 20-40°C a liz a rin e
0 p u rp u rin e
0,5 - 2 h pseudopurpurine MeOH/H2O
pH = 1,5 1
10 °C,
1
1. décom p tra ite m e n t 0 min.
5 p
le xa tio n a c id e H<0
OD / mg

1. RT
0 27

p ré c u rs e u r p ré c u rs e u rs
d'alizarine de lucidine
0.
5

anthragallol

0 .0

5 10 15 20 25
Sanyova J., Reisse J.(2006): Development of a mild method for the extraction of anthraquinones from their aluminium
complexes in madder lakes prior to HPLC analysis. Journal oet f Cmulptusral Heritage 7 (2006) 229–235
 Study of the historical sources
Workshop of Dirk Bouts, Christ Crowned with Thorns (NG
712)
Kermes lake was used in the red glaze and a madder lake in the pink
underpaint. In addition to the expected kermes dyestuff components,
ellagic acid was detected in the glaze suggesting extraction of the kermes
dydestuff from silk - ellagic acid is a component of oak galls (and a
breakdown product of many tannins).

Cross-section of
red paint,
showing red
140 lake in both red psp + Me
glaze and pink
120 underpaint
© The National Gallery, London
100

ea = ellagic acid
Absorbance / mAU

80

ka ka + Me ka = kermesic acid
60
al = alizarin
ea al pu
pu = purpurin
40
(psp) psp =
20
pseudopurpurin

0
50 60 70 80 90 100 110 120 130 1 40 150 16 0 1 70
 Reconstitutions
Kermes Al / Ca
Kermes Al / Ca / S

Triptyque de la Patience de Job, school Hieronymus Bosch, Groeningenmuseum Brugge

Sanyova J. (2008), Mild extraction of dyes by HF in routine analysis of historical paint micro-samples. Microchimca Acta ((361-370)
 Reconstructions
Kermes

Madder

purpurin

pseudopurpurin + munjistin (?)

2000 255 nm
320 nm (x 2)
Abs [mAU]

flavokermesic + kermesic acids

alizarin 495 nm (x 3)
HPLC
255nm

A070123_03, P168.060, Mlynica, volet ouvert, St Nicholas

benzoic acid

1000
inc "RT 27" Kermes

madder

0
St. Nicolas altarpiece, 20 30 40 50 60
Mlynica (Sk), 1500 Time [min]
Fading - xenotest
Lightfastness of lake powder
increase with the
concentration of dyestuffs
complexed (M/L)

The saturation of
the color increase
with concentration
of Al

=>
compromis
between
lightfastnes and
saturation of color
 Fading
8 , 3
2
Grade of lightfastnes : liz
A
fonction of the dye concentration
7
1
ui1 Pu
Q
6
4,7 1
A liz liz A
grade

5
liz8 liz5
A A
ui2
Q 3
4 6 ,
Aliz P u2

3
0 100 200 300 400 500 600 700
mg aliz / g laque (calc)
« Case studies »
Sedes Sapientiae, Liège (private col.), 1260-70
purpurin

1000
HPLC
pseudopurpurin
Madder
alizarin (Rubia tinctorum L.)
SEM-EDX

Abs [mAU]
500 P
b
S

Cl
Al Ca

Pb
Cloak 0 255 nm
Madder (Rubia tinctorum L.) 20 30 40 50
(Al, S,Ca) Time [min]
St Bishop and donator (privat collection),
1360(?) HPLC
Madder
(Rubia tinctorum L.)
Kermes
(K.vermilio Planch.)
Lac
(Kerria lacca Ker.)

SEM-EDX

Bishop’s red
mantel
Madder
Kermes
Lac Substrat:
(Al,S,Ca) Al, S, Ca
 Pre-eyckian painting, Namur, 1400
49 - bord de l'aile de l'ange, rouge fonce HPLC:
pu
al Rubia tinctorum L.
10 Porphyrophora
polonica (?)
Abs [mAU]
carminic acid
SEM-EDX :
0 alizarin purpurin
Al / Ca / S
HgS,
255 nm
-10
495 nm (x 3)
PbCO3.P
b(OH)2
20 30 40 50
Annonciation Time [min]

Coupe stratigraphique UV Vis

Projet « Peinture pré-Eyckienne
»
 Reliquary of St Maurice, Namur, 1400
Pantalon rose
HPLC:
20
benz.ac. ka

10

Abs [mAU]
fk

255 nm
495 nm
-10
20 30 40 50
Time [min]
HPLC:
SEM-EDX : Kermes (K.vermilio Planchon)
Cl Au Cl

(two differentes lakes)

Au
Ca Al Ca
S Au
K O
P
Au C
C
O S
K
Al
b
S SEM-EDX :
Au
K Ca
Cl Ca K
O
Pb
Cl Al, S,Cl,K, Pb
K Au K

0 2 4 0
C
2 4 0 2 4
Al / Ca / S et K/S
St. Nicolas’ Altarpiece, V. Lomnica, 1500-
1510
Vis UV Vis UV SEM

1 2
S

chromatogram 255 nm
C

HPLC Ca Al Ca
O

200 0 2 4 6

dehydro- 7-oxo-d e h y d r o
Fu ll S ca le 6 24 ct s

abietic acid
-
C ursor: 0.000

purpurin
abietic acid

alizarin
150

kermesic acid
Abs [mAU]

100 benzoic acid CERVENY

Red robe
SAT
2 Red robe
Kermes (Al) on
Madder (Al, S,Ca) 50
FIALOVY RUB PLASTA
Pin resin 2 1 Violet cloak, revers Violet cloak, revers
20
Kermes /Alumina 40
60
1 Fluorite, azurite
Time [min]
Pine resine
Sanyova Jana, Carmine. Kermes and Cochineal Lake Pigments . Postprint
of VIIth international seminar on restoration, Bratislava, 2008, p. 167-183
L.Lombard: Rebecca and Eliezer, Liège 1550
Vis
HPLC
At 255nm

UV

Lakes from:
- Dyers broom (Genista tinctoria L.)
- Madder (Rubia tinctorum L.)
- Kermes (Kermes vermilio Planchon)

Sanyova Jana, Mild extraction of dyes by hydrofluoric acid in routine analysis of historical paint micro-samples,
Microchim Acta 162, (on-line depuis 19 novembre 2007), 2008, p. 361–370.
L.Lombard:Coriolan reçoit sa mère,Liège 1550
Yellow pattern on green tente

255 nm
Tente verte et jaune (44) 350 nm genistein genistein
200
HPLC:
glycoside

Abs [mAU]
luteolin
apigenine
100
apigenin
apigenine glycoside
HPLC:
Dyers broom (Genista tinctoria L.) glycoside

SEM-EDX: 0 05_051019_12 HF
Al(OH)3, CaSO4, 20 30 40
(admixture of ochre and Pb-Sn yellow) Time [min]
Sanyova J., Saverwyns S. (2006) « Quelle technique dans l’atelier de Lambert Lombard ? » Scientia Artis 3, p.259-295
 Th.Van Loon: Adoration of shepherds, MRBAB, XVIIth C.
Vis
HPLC:

UV
SEM-EDX:

cochineal (Dactylopius coccus Costa)

brazil wood (Caesalpiniaces sp.)
dha and 7- (Pine resin)
oxo-dha
 Master of Trebon, 1380, NG in Prague.
B UV

Vis

679.53
3324.96

3067.88

2930.02

1652.98

837.73
2855.35

1558.21

1447.71
1412.07

1109.23
µ-FTIR
0.8

Amid I
Amid II
0.7

OH
0.6

CH2, CH3
Absorbance Units
0.5

UV6000-
Al-O
75 495nm
HPLC 75
0.4

P200.013
05_100521_03
pp aliz pu 495nm
0.3

.dat
Retention Time

36.2
0.2

CaCO3
29,5
50 50

42.4
0.1
0.0

mAU

mAU
3500 3000 2500 2000 1500 1000
Wavenumber cm-1

25 25
C:\JANA\Dossiers\2010_10667_Trebon,Prague\TR\08_110104_13 200.013 grain rouge.4 08_110104_13 200.013 grain rouge HYPERION 04/01/2011

Page 1/1

Two kinds of madder lake 0 0

+ azurite 10 20 30 40 50 60

+ charcoal, lead white (modeling)

Minute
s
A. Gierymski: Dama rokokowa, 1870-1901
m AU Alizarin

SEM-EDX
25

HPLC

HCl, 255nm
20

Xanthopurpurin,
Al(OH)3, Purpurin

BaSO4
15

Rhamnazin
10

Lawson
5

Madder (Rubia tinctorum L.)

5 10 15 20 25 30 35 40 min

Yellow lake (Rhamnus or 50

Purpurin
50

Munjistin
quercus ?) 40 HF, 255nm pseudopurpurin 40

(Al, Ca / Ba) 30 30

20
Alizarin 20

quercitrin
10 10

U
Benzoic

mAU
A
m
0 0

acid
-10 -1 0

-20 -2 0

-3 0
-30

-4 0
-40

-5 0 -5 0
16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48
50 M in u te s

E. Doleżyńska-Sewerniak, W. Nowik, J. Sanyova: Analytical investigation of organic dyestuffs in glazes of Aleksander Gierymski’s paintings from 1870-1901
 Synthetics dyestuffs, 1903-1934
Blue pigment from the collection of
prof. Gottfried Kallstenius,
“Royal Academy of Fine Arts in
Stockholm” (1903 - 1934)
SEM-EDX : BaSO4, Na, Ca, C.
MRS: too much fluorescence -identification impossible
HPLC: 5,5'-indigodisulfonic acid
indigo disulfonic acid 255 nm

200

Abs [mAU]

100

06222_05, F157, substrat :

indigo sulfonic acid

BaSO4
0

20 40
indigo-carmin / BaSO4 Time [min]

Sanyova Jana, Mild extraction of dyes by hydrofluoric acid in routine

C.I. Natural Blue 2, Acid Blue 74, analysis of historical paint micro-samples, Microchim Acta 162, (on-line
depuis 19 novembre 2007), 2008, p. 361–370.
Thanks
 al-pr Analytical protocol

1
<0,1mg
sample + 20µL
Au

DM F
quercitrin

HF
unk al-pr
lc-pr pp+mu al
ru-pr pu
ru
unk27
2
+/ACN
HF 4N ->
discoloration

3
al
pu
Evap. to
unk unk mu +unk
dryness (N2)
HCl ru unk27

4
redissolution
10 20 30 40 50 in DMF/H20
Time (min)

5
HPLC analyse

Red-brown organic pigment used by Nils Masson Mandelgren, between 1850-1880

 Site of complexation

Al+++
O O
- Al+++ O-
O
O- OH

diphenolate Keto-phenolate
Complexe alizarin-aluminium
(Fierz-David et Rutishauer, 1940)

O O

Al
Complexe alizarin-aluminium
(Kiel et Heertjes, 1963)
Complexe alizarin-aluminium
(Soubayrol, 1996)
4-

H
-
O
- -
O Al O Al O - 4 Na+
O
O