',AR.CHIVES.
'"..
FISHERIES AND MARINE SERVICE
Tr.anslat1on Series NO. 4174
Use of the IATROSCAN TH 10 for the separation and determination
by thin-layer -chromatography of some components in fatty substances
,•
by E. Gantois, F. Mordret, and
N. Le Barbanchon
Original title: Utilisation de l'appareil'IATROSCAN TH 10 pour la separation et
le dosage par chromatographie sur couche mince de quelques constituants
des corps gras
From: Rev. Fr. Corps Gras
24(3): 167-169, 1977
Translated by the Translation Section .
Departmn
of the Environment
Department of the Environment
Fisheries and Marine Service
Halifax Laboratory
. Halifax, N.S..
'
6
pages typescript
1977
.
1088921
p. 167.
d-
11
USE OF THE IATROSCAN TH 10
FOR THE SEPARATION AND DETERMINATION
BY THIN-LAYER CHROMATOGRAPHY
OF SOME COMPONENTS IN FATTY SUBSTANCES
LABORATORY REPORT
E. GANTOIS, F. MORDRET, N. LE BARBANCHON
Institut des--cieps---gras—z-z-Parts-.
RFCG 77-14
The IATROSCAN TH 10 apparatus is purported to enable the automatic,
rapid and quantitative detection of compounds that may be separated by thinlayer chromatography. We therefore decided to apply this new method to a
few mixtures of components norm.i; fractioned by TLC (paf•tial glycerides,
unsaponifiables) and present in fatty substances.
APPARATUS OPERATING PRINCIPLE
With this apparatus, compounds previously separated by thin-layer
chromatography under particular conditions are detected
a flame
ionization system,(1).
---
The chromatography is done on fine quartz rods (0 -:.0.8rrni,
L:152 mr•), covered with a layer of silica 75p thick.
marods'',
Ten of these "chro-
mounted on a metal frame, receive a punctual deposit of the solution
to be analyzed, then the entire set is placed vertically in a developing tank.
---
The detection is done in the apparatus itself, which includes
a holder upon which the rods are placed one at a time after the chromatography.
This holder moves at a uniform speed so that the rods are subjected to the
detector flàme at a constant speed (Figure 1). An automatic mechanical
.eystee-enables the treatment in series of all ten rds.
theis move through the apparatus is regulated
fn
\
The speed at which
advance (set of gears),
7
while the position of the burner ln relation to the'),:èds way be adjusted at
will.
The apparatus is also equiped with a device to record the'integration
1088921
p. 167.2
curve or may be connected to an electronic integrator.
-
1M.
The chromarods are regenerated and activated during the
analysis and the same process may be applied to new ohremerods.
Rods may be
used up to one hundred times.
FIGURE 1
IIA6RAM
FIE-CIPERA1 l'NG FRJN
1-
Collecting electrode
2-
Chromarod
3-
Burner
4-
Signal amplifier
5-
Integration amplifier
6-
Integrator
7-
Recorder
RESULTS
Since the apparatus was available to us for a short time only,*
we tried to determine the repeatability of the method and compare its results
with those obtained through densitametry or by weighing.
* WebtisIttià thank the INTERSMAT Corporation for placing this apparatus at
our disposal.
1068921
p. 168.1
TABLE 1
REPEATABILITY TESTS
GLYCERIDES 1
SAMPLES
Composition of TG: 53
GLYCERIDES 11
MG: 47
TG: 75
MG:25
41.3
74.9
25.1
UNSAPONIFIABLE
Alcohols
Squalene Terp.)Ssitosterol
62.3
11.5
26.2
mixture
Results
58.7
No. of measurements 20
62.1
4
10.4
27.5
10
Standard
deviation
4.3
4.4
1.2
1.2
1.5
0.9
2.8
Variation
coefficient (le)
7.3
10.6
1.6 .
4.8
2.5
8.6
10.4
1.
METHOD REPEATABILITY
Tests were conducted with synthetic mono and trygliceride mixtures
and with mixtures containing unsaponifiable components (Table 1).
Repeatability is generally good, but results may be fairly different
from anticipated values (mixture 1) if the detector controls are not adjusted
quite correctly (flow H2 , movement of chromarods).
By using an electronic
integrator and optimizing the conditions of response a solid correspondence
may be obtained between the indications given by this method and the actual
composition of the mixtures.
Similar results wervieently published (2)
for the analysis of cholesterol, stearic acid, tripalmitin, and cholesteryl
palmitate mixtures.
COMPARISON OF VARIOUS METHODS.
APPLICATION TO THE ANALYSIS
OF TUO INDUSTRIAL MONOGLYCERIDES (Figure 2).
FIGURE 2
INDUSTRIAL MONOGLYCERIDES
(left to right)
solvent front;
triglycerides; diglycerides; monoglycerides.
Two industrial monoglycerides (monelein and soya) were first
analyzed by silica column chromatography. These samples also underwent
•
1088921
p. 168.2
thin-layer fractioning under particular conditions of development (3 successive
solvents), so that the bands representing mono, di and triglycerides had
•
Rfs between 0.2 and 0.6, which is the desirable=(for densitometric
measuring. Results are shown in Table II.
A good concordance was recorded between the IATROSCAN and densitometry
for the soya monoglyceride. The difference in values found by column chromatograMy is probably due to the high level of polarity of certain glycerides
which are thus retained.
m
aece-ef-ealues
is not as good, and the result,' obtained through the IATROSCAN is very different
from that obtained through column chromatography.
was noted in each case.
The duration of the analysis
Results may be obtained very rapidly with the
IATROSCAN: ten determinations (similar or different) may be effected in 40
minutes.
TABLE II
---
-COMPARISON-OF-INDUSTRIAL-MONOGLICERIDLANALYSIS
RESULTS FROM IATReSCAN TH 10 AND TRADITIONAL METHODS
Method used
Soya monoglyceride
MG% DG% TG%
Rod Chromatography
and analysis with
43.4 47.2 9.4
Iatroscan TH 10
Variation Coefficient
Industrial Monooleine Duration of
analysis in
TG%
MG% DG%
hours
50
42.1
7.9
6.3
6.1
Plate Chromatography 44.0 45.2 10.8
and densitometry
Variation Coefficient
44.6 43.1
12.2
Column Chromatography 43 •4* 35.2* 9*
and weighing
50
3.3
6.9
5.1
42
*The highly unsaturated soya glycerides wasiUmmt
.4>f the co1umn aed-recovery was only 87.6%
0
44.1
tuly
Oh4Omin for
10 determinatio
4 h for 10
determinations
12.3
8
6 h for one
determination
1068921
p. 169.1
CONCLUSION
We feel this apparatus is of significant interest because it
provides a satisfactory solution (detection by flame ionization) to quantitative analysis problems in thin-layer chromatograptly. Its automatic design
enables the treatment of up to ten samples at a time and its connection
to an electronic integrator considerably reduces the total duration of
analysis in series, which may be reduced to only a few minutes.
It would be advisable that these preliminary results be confirmed
by further testing.
EXPERIMENTAL APPENDIX
1-
EeRtatellity
- --
TestAnalysis Conditions (Table 1)
Sample: mixture of monoglycerides and triglycerides (47%
Kodak _monol eine ver.__99% _pure) and _.53%__o
ve_oi_l_).
- --
Deposit: 1 to 2 pl of the preceding mixture 0.2% in chloroform.
---
Stationary phase: chromarod S.
- --
Mobile phase:
- - Glycerides: chloroform, benzene, formic acid (60-40-2).
Drying of rods 3 minutes at 1300C.
▪ Unsaponifiable: hexane - ethyl oxide, formic acid (70-30-1).
--- Analyzer adjustment
- - Flame
hydrogen 1.2 b (168 ml/min)
air 2 1/min
- - speed of rod passage through flame: 3.8 cm/s
---
2-
Recorder 10 mv -- unrolling 30 cm/min
Industrialtior Ilce
- --
alsisCormlitions (Table II)
Sample: industrial soya monoglycerides
Industrial monoleine.
--- Deposit and stationary phase: same as 1
---
Mobile phase: hexane, ethyl oxide, formic acid (60-40-1)
1 088921
p. 169.2
»MM.
Analyzer adjustment:
MO»
Flame: same as 1
speed of rod passage-filename: 2.9 cm/s
111. 10■ ••
Recorder: sanie as 1
BIBLIOGRAPHY
(1) T. OKUMURA, T. KADONO, A. IMO -- J. of
Chromatography 1975, 108, 329-336.
(2)M. TANAKA, T. ITOH, H. KANEKO. -- Yukagaku, 976, 25, 263.
(3) M. NAUDET, J. PASERO, S. BIASINI -- Rev. Franç. Corps Gras, 1965, 12, 525.
(Manuscript received February 17, 1977)