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IATROSCAN APPLICATION
MITSUBISHI CHEMICAL MEDIENCE2-8, Shibaura 4-Chome, Minato-ku, Tokyo 108-8559, Japan
http://www.medience.co.jp/
I A T _ A _ E V e r . 1 . 0
C O N T E N T
APPENDIX Ⅰ Published Scientific References
APPENDIX Ⅱ IATROSCAN Application Data Application No. Page
Keys to Analytical Procedure ………………………………………………….… 1A 46
Type Analysis for Heavy Oil ……………………………………………………... 2A 60
Analysis of Biodegraded Crude Oil by Iatroscan ……………………………… 11-Ⅱ 65
Analysis of Lipids by Iatroscan …………………………………………….....… 5 70
Analysis of Lipids by Iatroscan ………………………………………………..... 5-Ⅱ 77
Analysis of Lipids by Iatroscan (Marine Products) ......................................... 23 88
Chromarod-SIII Treatment Procedure for FPD Analyzing
Phosphorous Compound ………………………………………………………...
4A
98
Analysis of Glyceride isomers using Boric acid-Impregnated Chromarods … 12 104
Analysis of Triglyceride Molecular Species using Silver nitrate-Impregnated
Chromarods ...................................................................................................
16
111
Lipid Analysis using Copper Sulphate Impregnated Chromarods ................ 22 120
Experimental Analysis for Infinitesimal Components Contained in Main
Ingredients (Detection of Infinitesimal Phospholipid in Edible Oils) …………
20
126
Tracing of Reaction with Enzymatic Experimental Reaction by Iatroscan ....... 13 133
Separation of Isomers and Derivatives .......................................................... 9 137
Analysis of Surface Active Agents by Iatroscan ............................................. 21 146
Analysis of Polymer Additives by Iatroscan ................................................... 24 156
Environment and Health Friendly Developing Solvents for Iatroscan ........... 26 166
Chromatograms .............................................................................................
Dye(Food dye, Napthol quinone, Azo dye), Hormons (Pregnandiol),
Ginseng Saponin, Liquid Crystal, Capsaicine, Cosmetic Cream,
Rubber Antioxidant, Polymer
175
APPENDIX Ⅰ
Published Scientific References
General InformationThe IATROSCAN is an automatic detector that performs quantitative analysis onorganic mixtures separated on thin layer chromatography (TLC) and detected byHydrogen Frame Ionization System (FID). The separation of components isperformed on an exclusive thin layer chromatography media (CHROMAROD) inthe same manner of normal phase TLC. Its FID system has high sensitivity foralmost all organic components. The detection selectivity applies to a wide varietyof samples. Particularly, lipid analysis, monitoring reaction rates in organicsynthesis and fractionation of residues of crude oil and asphalt are easilyanalyzed with the IATROSCAN. Ten CHROMARODs are held in a single rackallowing up to 10 samples to be applied at a time. All 10 are processedsimultaneously and detected in series to obtain the final results quickly.
Principle of OperationWhen a sample(s) is developed and separated on the CHROMAROD (thin layerquartz rod) and scanned directly into the Hydrogen Flame at the rated speed,organic components separated on this thin layer surface are ionized by theenergy of Hydrogen Flame. The ions generated are charged both negative andpositive. The negative ions (-) flow to the Burner and the positive ions (+) flow tothe Collector Electrode due to the electric field loaded between the FID electricpoles {Burner positive (+) and Collector negative (-)}. These ion currents flowbetween the Burner and the Collector proportionally to the mass of componentsbeing ionized in the Hydrogen Flame. The ion current is amplified by the FIDcircuit, and the components are quantitatively measured and recorded by thedata processing unit.
Photomultiplier tube
Date Processing Unit
Interference FilterFIDAmmeter
Collector Electrode
Light GuideHydrogen
FPDAmmeter
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12 / 188
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prawn Penaeus japonicus.” Biochimie (77) 190-193, 1995.
237. Cavaletto,J.F., Nalepa,T.F., Dermott,R., Gardner,W.S., Quigley,M.A., Lang,G.A.,
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spp. (Amphipoda) from lakes Michigan and Ontario.” Can J Fish Aquat Sci.
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238. Kim,J.K., Chung,G.H., Rhee,J. S., “Concentration of bovine plasmalogen
using phospholipase A1 in two-phase system.” Biotechnol Tech. (13) 589-593,
1999.
239. Garcia-Ayuso,L.E., Velasco,J., Dobarganes,M.C., Luque de Castro, M.D.,
“Double use of focused microwave irradiation for accelerated matrix hydrolysis
and lipid extraction in milk samples.” International Dairy Journal (9)
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240. MacFarlane,R.B., Norton,E.C., “Nutritional dynamics during embryonic
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development in the viviparous genus Sebastes and their application to the
assessment of reproductive success.” Fish Bull. 97(2) 273-281, 1999.
241. Bergen,B.J., Quinn,J.G., Parrish, C.C., “Quality-assurance study of marine
lipid-class determination using Chromarod/Iatroscan thin layer
chromatography-flame ionization detector.” Environ Toxicol Chem. 19(9)
2189-2197, 2000.
242. Pamies-Andreu,E., Garcia-Lozano,R., Palmero-Palmero,C., Garcia-Morillo,S.,
Alonso-Arcas,A., Stiefel,P., dela Fuente,J.C., Villar,J., “Genetic variation in the
beta-3-adrenergic receptor in essential hypertension.” Life Sci.(67) 391-397,
2000.
243. Maswadeh,H., Hatziantoniou,S., Demetzos,C., Dimas, K., Georgopoulos,A.,
Rallis, M., “Encapsulation of Vinblastine into New Liposome Formulations
Prepared from Triticum (Wheat Germ) Lipids and its Activity Against Human
Leukemic Cell Lines.” Anticancer Res. 20(6B) 4385-4390, 2000.
244. Hsu,H-C., Lee,Y-T., Chen,M-F., “Effect on n-3 fatty acids on the composition
and binding properties of lipoproteins in hyper-triglyceridemic patients.” Am J
Clin Nutr. (71) 28-35, 2000.
245. Fonollosa,J., Marti,M., de la Maza,A., Sabes,M., Parra,J. L., Coderch, L.,
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Langmuir. 16(11) 4808-4812, 2000.
246. Goutx,M., Momzikoff,A., Striby,L., Andersen,V., Marty,J.C., Vescovali,I.,
“High-frequency fluxes of labile compounds in the central Ligurian Sea,
northwestern Mediterranean.” Deep-Sea Research I (47) 533-556, 2000.
247. Nates,S.F., McKenney Jr,C.L., “Onto genetic changes in biochemical
composition during larval and early postlarval development of
Lepidophthalmus louisianensis, a ghost shrimp with abbreviated development.”
Comp Biochem Physiol. B (127B) 459-468, 2000.
248. Nalepa,T.F., Hartson,D.J., Buchanan,J., Cavaletto,J.F., Lang,G.A., Lozano,S.J.,
“Spatial variation in density, mean size and physiological condition of the
holarctic amphipod Diporeia spp. in Lake Michigan.” Freshwater Biology
(43) 107-119, 2000.
249. Dominguez,C., Coderch,L., Erra,P., Bayona,J.M., Palet,D., “Mineral oil
detection in spinning oil formulation and wool top dichloromethane extract by
TLC/FID.” AATCC Review 1(7) 6-8, 2001.
250. Ylitalo,G.M., Matkin,C.O., Buzitis,J., Krahn,M.M., Jones,L.L., Rowles,T.,
Stein,J.E., “Influence of life-history parameters on organochlorine
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concentrations in free-ranging killer whales (Orcinus orca) from Prince William
Sound, AK.” Sci Total Environ. (281) 183-203, 2001.
251. Phillips,K.L., Jackson,G.D., Nichols, P.D., “Predation on myctophids by the
squid Moroteuthis ingens around Macquarie and Heard Islands: stomach
contents and fatty acid analyses.” Mar Ecol Prog Ser. (215) 179-189, 2001.
252. Graeve,M., Dauby,P., Scailteur,Y., “Combined lipid, fatty acid and digestive
tract content analyses: a penetrating approach to estimate feeding modes of
Antarctic amphipods.” Polar Biol. (24) 853-862, 2001.
253. Lopez,O., Cocera,M., Parra,J.L., dela Maza, A., “Influence of the alkyl chain
length of alkyl glucosides on their ability to solubilize phosphatidylcoline
liposomes.” Coll Surf. A (193) 221-229, 2001.
254. Uno,S., Yun,J.H., Kaneniwa,M., Koyama,J., Yamada,H., Ikeda, K., “Lipid
class armed fatty acid composition of mussel, Mytilus trossulus, in Vancouver
Harbour.” PICES Scientific report (16) 43-47, 2001.
255. Jeffs,A.G., Phleger,C.F., Nelson,M.M., Mooney,B.D., Nichols,P.D., “Marked
depletion of polar lipid and non-essential fatty acids following settlement by
post-larvae of the spiny lobster Jasus verreauxi.” Comp Biochem Physiol.
(131A) 305-311, 2002.
256. Engel,E., Lombardot,J-B., Garem,A., Leconte,N., Septier,C., Quere,J-L L.,
Salles,C. “Fractionation of the water-soluble extract of a cheese made from
goats' milk by filtration methods: behaviour of fat and volatile compounds.”
International Dairy Journal (12) 609-619, 2002.
257. Scott,K.D., Fahraeus-Van Ree,G.E., Parrish,C.C., “Sex differences in hepatic
lipids of Toxaphene-expased juvenile yellowtail flounder (Pleuronectes
ferrugineus Storer).” Ecotoxicol Environ Saf. 51(3) 168-176, 2002.
258. Murphy,K.J., Mooney,B.D., Mann,N.J., Nichols,P.D., Sinclair,A.J., “Lipid,
FA, and sterol composition of New Zealand green lipped mussel (Perna
canaliculus) and Tasmanian blue mussel (Mytilus edulis).” Lipids 37(6)
587-595, 2002.
259. Yohannes,K., Dalton,C.B., Halliday,L., Unicomb,L.E., Kirk,M., “An outbreak
of gastrointestinal illness associated with the consumption of escolar fish.”
Commun Dis Intell. (26) 441-445, 2002.
260. Pinturier-Geiss,L., Mejanelle,L., Dale, B., Karlsen,D.A., “Lipids as
indicators of eutrophication in marine coastal sediments.” J Microbiol
Methods 48(2-3) 239-257, 2002.
261. Mecozzi,M., Amici,M., Romanelli,G., Pietrantonio,E., Deluca,A., “Ultrasound
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extraction and thin layer chromatography-flame ionization detection analysis of
the lipid fraction in marine mucilage samples.” J Chromatogr.A (963) 363-373,
2002.
262. MacFarlane,R.B., Norton, E.C., “Physiological ecology of juvenile chinook
salmon (Oncorhynchus tshawytscha) at the southern end of their distribution,
the San Francisco Estuary and Gulf of the Farallones, Calfornia.” Fish Bull.
100(2) 244-257, 2002.
263. Pedersen,S.A., Storm, L., “Northern shrimp (Pandalus borealis) recruitment
in west greenland waters part II. Lipid classes and fatty acids in Pandalus
Shrimp Larvae: Implications for survival expectations and trophic
relation-ships.” J Northw Atl Fish Sci. (30) 47-60, 2002.
264. Dominguez,C., Jover,E., Garde,F., Bayona,J.M., Erra,P., “Characterization of
supercritical fluid extracts from raw wool by TLC-FID and GC-MS.” JAOCS
80(7) 717-724, 2003.
265. Dominguez,C., Jover,E., Bayona,J.M., Erra, P., “Effect of the carbon dioxide
modifier on the lipid composition of wool wax extracted from raw wool.”
Analytica Chimica Acta (477) 233-242, 2003.
266. Attar-Bashi,N.M., Orzeszko,K., Slocombe,R.F., Sinclair,A.J., “Lipid and FA
analysis of canine prostate tissue.” Lipids 38(6) 665-668, 2003.
267. Copeman,L.A., Parrish,C.C., “Lipids classes, fatty acids, and sterol in
seafood from Gilbert Bay, southern Labrador.” J Agric Food Chem. 52(15)
4872-4881, 2004.
268. Caradec,S., Grossi,V., Gilbert,F., Guigue,C., Goutx,M., “Influence of various
redox conditions on the degradation of microalgal triacylglycerols and fatty acid
in marine sediments.” Organic Geochemistry (35) 277-287, 2004.
269. Coderch,L., Fonollosa,J., Marti,M., Parra,J. L., “Ceramides from wool wax.”
JAOCS (81) 897-898, 2004.
270. Goniotaki,M., Hatziantoniou,S., Dimas,K., Wagner,M., Demetzos,C.,
“Encapsulation of naturally occurring flavonoids into liposomes:
physicochemical properties and biological activity against human cancer cell
lines.” J Pharm Pharmacol. (56) 1217-1224, 2004.
271. Smidrkal,J., Cervenkova,R., Filip, V., “Two-stage synthesis of sorbitan esters,
and physical properties of the products.” Eur J Lipid Sci Technol. (106)
851-855, 2004.
272. Vikbjerg,A. F., Mu,H., Xu,X., “Monitoring of
monooctanoylphosphatidylcholine synthesis by enzymatic acidolysis between
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soybean phosphatidilcholine and caprylic acid by thin-layer chromatography
with a flame ionization detector.” J Agric Food Chem. (53) 3937-3942, 2005.
273. Linder,M., Fanni,J., Parmentier,M., “Proteolytic extraction of salmon oil and
PUFA concentration by lipases.” Mar Biotechnol. (15) 70-76, 2005 .
274. Copeman,L.A., Parrish,C.C., Brown,J.A., Harel,M., “Effects of
docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth,
survial, lipid composition and pigmentation of yellowtail flounder (Limanda
ferruginea): a live food enrichment experiment.” Aquaculture (210) 285-304,
2002.
2.2 Fats & Oils275. Gantois,E.,Mordret,F.,LeBarbanchon,N., “Utilisation de l'appareil Iatroscan
TH-10 pour la separation et le dosage par chromatographie sur couche mince de
quelques constituants des corps gras.” Rev.France Corps Gras (24) 167, 1977.
276.Ackman,R.G.,Eaton,C.A.,Crewe,N.F.,Nash,D.M.,Loew,F.M.,Schiefer,B.,Forsyth,
G.W.,Olfert,E.D.,Laxdal,V., “Longer term comparisons of clinical,biochemical
and physiological effects of lard-corn oil and partially hydrogenated herring oil
as major dietary fats of cynomolgus monkeys.” Fisheries and marine service
technical report No.808 Sept., 1978.
277. Isobe,T.,Seino,H.,Watanabe,S., “Study on the lipase hydrolysis of castor oil
(I),Isolation of glycerol triricinoleate from castor oil.” ACS/CSJ Chemical
congress, 1979.
278. Pore,J,,Noel,J.P., “Controle des bains de nettoyage des vetements de peaux.”
Revue technique des industries du cuir (72 ) 65, 1980.
279. Houis,J.P.,Assimacopoulos,E.,Rasori,I.,Pore,J.,“Application de la
chromatographie couche mince (TLC-FID) au controle des emulsion
industrielles.” Revue francaise des corps gras (27) 61, 1980.
280. Tanaka,M., Itoh,T. & Kaneko,H., “Quantitative determination of isomeric
glycerides, free fatty acids and triglycerides by thin layer chromatography-flame
ionization detector system.” Lipids 15(10) 872, 1980.
281. Sebedio,J.L. & Ackman, R.G., “Chromarod-S modified with silver nitrate for
the quantitation of isomeric unsaturated fatty acid.” J.Chromatogr. Sci (19)
552, 1981.
282. Ranny,M., Sedlacek,J., Mares,E., Svoboda,Z. & Seifert,R., “Quantitative
Analyse von molekular destillierten Monoacylglycerolen mittels der
Dunnschichtchromatographie und Flammenionisationsdetektor (TLC-FID).”
Seifen-Ole-Fette-Wachse 109(8) 219～224, 1983.
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283. Tatara,T., Fujii,T,. Kawase,T & Minagawa,M., “Quantitative determination of
tri-, di-, monooleins and free oleic acid by the thin layer chromatography-flame
ionization detector system using internal standards and boric acid impregnated
chromarod.” Lipids 18(10) 732, 1983.
284. Wahren,H.L., Herslof,M & Larsson, K., “A comparison of the phase behaviour
of the monoolein isomers in excess water.” Chemistry and Physics of Lipids
(33) 211, 1983.
285.Sebedio,J.L. & Ackman,R.G., “Hydrogenation of a menhaden oil: Ⅱ.Formation
and evolution of the C20 dienoic and trienoic fatty acids as a function of the
degree of hydrogenation.”JAOCS 60(12) Dec., 1983.
286. Morita,S., Narita,H., Matoba,T. & Kito,M., “Synthesis of triacylglycerol by
lipase in phosphatidylcholine reverse micellar system.” JAOCS (61) 1571,
1984.
287. Freedman,B., Pryde,E.H. & Kwolek,W.F., “Thin layer chromatography/flame
ionization analysis of transestrified vegetable oils.” JAOCS (61) 1215, 1984.
288. Kramer,J.K.G., Thompson,B.K., Farnworth,E.R., Varnworth,E.R., “Variation
in the relative response factor for triglycerides on Iatroscan Chromarods with
fatty acid composition and sequence of analyses.” J. Chromatogr. (355) 221,
1986.
289. Sebedio,J.L., Septier,C., Grandgirard,A., “Fractionation of commercial frying
oil samples using sep-pak cartridges.” JAOCS (63) 1541, 1986.
290. Hara,K., Cho,S.Y., Fujimoto,K., “Measurement of polymer and polar
material content for assessment of the deterioration of Soybean oil due to heat
cooking.” YUKAGAKU (38) 463, 1989.
291. Petersson,B., “Determination of triglycerides according to their degree of
ucnsaturation using mercury(Ⅱ) acetate adducts and thin-layer
chromatography with flame-ionization detection.” J. Chromatogr., (242) 313,
1982.
292. Yamane,T., Hoq,M.M., Itoh,S., “Glycerolysis of fat by lipase.” J. Jpn. Oil
Chem. Soc. (35) 625, 1986.
293. Sebedio,J.L., “Chromatographic techniques applied to the analyses of heated
fats and oils.” Chromatography and analysis, October 9, 1991.
294. Thankappan,T.K., Gopakumar,K., “A rapid method of separation and
estimation of squalene from fish liver oils using Iatroscan analyser.” FISHERY
TECHNOLOGY 28, 1991.
295. Matori,M., Asahara,T., Ota,Y., “Positional specificity of microbial lipases.”
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Journal of Fermentation and Bioengineering (72) 397, 1991.
296. McNeill,G.P., Shimizu,S., Yamane,T., “High-yield enzymatic glycerolysis of
fats and oils.” JAOCS (68) 1, 1991.
297. McNeill,G.P., Yamane,T., “Further improvements in the yield of
monoglycerides during enzymatic glycerolysis of fats and oils.” JAOCS 68(1) 6,
1991.
298. Banerjee,A.K., Ratnayake,W.M.N., Ackman,R.G., “Enhanced response of
more highly unsaturated lipids on thin-layer chromatography-Iatroscan flame
ionization detection after exposing the developed Chromarods to iodine vapour.
J. Chromatogr. (319) 215, 1985.
299. Sebedio,J.L., Astorg,P.O., Septier,C., Grandgirard,A., “Quantitative analyses
of polar components in frying oils by the Iatroscan thin-layer
chromatography-flame ionization detection technique.” J. Chromatogr. (405)
371, 1987.
300. Chandrasekhara,T.R., Vijayalakshmi,P., Gangadhar,A., Subbarao,R.,
Lakshminarayana,G., “Chromatographic methods for the determination of
monomer, dimer and trimer fractions in dimer fatty acids.” J. Chromatogr.
(466) 403, 1989.
301. Koike,S., Yokoo,M., Nabetani,H., Nakajima,M., “Membrane separaion of fats
and oils in organic solvents.” Progress in bioseparation engneering 1992, The
Society of Chemical Engineers, Japan. 84, 1993.
302. Hara,K., Hasegawa,K., Endo,Y., Fujimoto,K.,“Polymer and polar material
content determination as basis for assessing thermally oxidative deterioration of
soybean oil for potato heat cooking.” J. Jpn. Oil Chem. Soc. (YUKAGAKU)
43(1) 57, 1994.
303. Sahashi,Y., Ishizuka,H., Koike,S., Suzuki,K., “Membrane separation of
polyunsaturated fatty acids-rich acylglycerods from fish oil.”YUKAGAKU (43)
116, 1994.
304. Marquez-Ruiz,G., Perez-Camino,M.C., Carmen,M., “Evaluationof
susceptibility to oxidation of linoleyl derivatives by thin-layer chromatography
with flame ionization detection.” J. Chromatogr. (A 662) 363, 1994.
305. Rios,J.J., Perez-Camino,G., Marquez-Ruiz., Dobarganes,M.C., “Isolation and
characterization of sucrose polyesters.” JAOCS 71(4) 385, 1994.
306. G.Marquez-Ruiz., M.C.Perez-Camino.,J.J.Rios., M.C.Dobarganes.,
“Characterization of sucrose polyesters-triacylglycerols mixtures.” JAOCS
71(9) 1017, 1994.
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307. Sathyanarayana,M.N., “Compositional analysis of partial glycerides.”
PAINTINDIA 44(8) 31, 1994.
308. Peyrou,G., Rakotondrazafy,V., Mouloungui,Z.,Gaset,A., “Separation and
quantitation of mono-,di,and Triglycerides and free oleic acid using thin-layer
chromatography with flame-ionization detection.” Lipids 31(1) 27-32, 1996.
309. Liang J-H ,Hwang L.S., “Ethylation Reaction of Squid Visceral Oil.” Food
Science 24(4) 408-418, 1997.
310. Cruzian J.L., Inhamuns A.J., y Barrera-Arellano, “Determinacion de
compuestos polares por TLC-FID en aceites refinado y semi-hidrogenado de soja
sometidos a calentamiento prolongado.” Grasas y Aceites (48) 148-153,
1997.
311. Marquez-Ruiz G, Guevel G., Dobarganes M.C., “Applications of
chromatographic technigues to evaluate enzymatic Hydrolisis of oxidized and
polymeric triglycerides by pancreatic lipase in virto.”
JAOCS 75(2) 119-26, 1998.
2.3 Carbohydrates312. Kondo,Y., “Partial benzoylation of methyl 4,6-O-benzylidene-α-and
β-D-glucopyranosides, and 1,5-anhydro-4,6-O-benzylidene-D-glucitol with
benzoic anhydride in chloroform.” Agri. Biol. Chem.41(10) 2089, 1977.
313. Kondo,Y.,“A reinvestigation of partial benzoylation of 1,5-anhydro
4,6-O-benzylidene-D-glucitol.” Agri. Biol. Chem.41(12) 2481, 1977.
314. Kondo,Y., “Partial tosylation of 1,5-anhydroxylitol.” Carbohydrate
Research. (103) 154, 1982.
315. Kondo,Y.,“Selective benzoylation of methylα-andβ-D-xylopyranoside.”
Carbohydrate Research. (107) 303, 1982.
316. Kondo,Y.,“Partial sulfonylation of methyl α-and β-D-xylopyranoside.”
Carbohydrate Research. (110) 339, 1982.
317. Kondo,Y., “Selective benzoylation of 1,5,-anhydro-D-galactitol.”
Carbohydrate Research. (121) 324～327, 1983.
318. Vydra,C., Ranny,C. & Sedmera,B., “Partially acetylated sucrose.”
Collection on Czechoslovak Chem Commun. (50) 1040, 1985.
319. Kondo,Y., “Partial benzoylation of 1,5-anhydroxylitol.” Carbohydrate
Research (111) 325, 1983.
320. Kondo,Y., “Selective benzoylation of 1,5-anhydro-L-arabinitol.”
Carbohydrate Research (134) 167, 1984.
321. Esaiassen,M., Overbo,K., Olsen,R.L., “Analysis of
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N-acetyl-chitooligosaccharides by the Iatroscan TLC/FID system.”
Carbohydrate Research (273) 77-81, 1995.
322. Yokoyama,S., Uematsu,J., Inoue,K., Katoh,T.,& Sanada,Y., “Chemical
structure of coal hydrogenation liquidscompound type estimation by TLC-FID.”
Proceedings international conference on coal science. P.465, 1981.
323. Poirier,M.A. & George, A.E., “Rapid method for determination of malthene
and asphaltene content in bitumen, heavy oils, and synthetic fuels by pyrolysis
TLC.” J. Chromatogr. Sci. (21) 331, 1983.
324. Poirier,M.A. & George, A.E., “A rapid method for hydrocarbon type analysis of
heavy oils and synthetic fuels by pyrolysis thin layer chromatography.”
Energy Sources 7(2) 151, 1983.
325. Boden,H. & Roussel, R., “A method for determining polycyclic aromatic
hydrocarbons for coal tar pitch volatiles.” 112th AIME Annual Meeting,
Atlanta, 6-10. March, 1983.
326. Selucky,M.L., “Quantitative analysis of coal-derived liquids by thin-layer
chromatography with flame ionization detection.” Anal. Chem. 55(1) 141,
1983.
327. Obuchi,A., Aoyama,H., Ohi,A., Ohuchi,H.., “Application of thin-layer
chromatography with flame ionization detection to the characterization of
organic extracts from diesel exhaust particulated.” J. Chromatogr. (288) 187,
1984.
3. Industrial Chemical3.1 Heavy Oil and Coal Liquefaction Products
328. Yokoyama,S., Umematsu,J., Inoue,K., Katoh,T & Sanada, Y., “Estimation of
compound classes in coal hydrogenation liquids by thin-layer chromatography.”
Fuel (63) 984, 1984.
329. Kaschani,D.T., “DGMK-Project 355 Entwicklung eines schnellen Verfahrens
zur Bestimmung von Kohlenwasserstofftypen in hoch-und nichtsiedenden
Mineralo-Fraktionen.” Deutsche gesellschaft fur mineralolwissenschaft und
kohlechemie e.v., 1986.
330. Yamamoto,Y., “Analysis of heavy oils by FID-TLC. (part 4) A rapid method
for hydrocarbon type analysis of heavy oils. Sekiyu Gakkaishi (29) 15, 1986.
331. Yamamoto,Y. & Akutsu,H., “Analysis of heavy oils by FID-TLC. (part 5) A
rapid method for determination of olefin contents.” Sekiyu Gakkaishi (29) 38,
1986.
332. Yoshida,R., Miyazawa,M., Yoshida,T., Ishizaki,K., Shinriki,N. & Maekawa,Y.,
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“Upgrading of coal-derived liquids. 3. characterization of upgraded liquids by
thin-layer chromatography combined with flame-ionization detection.” Fuel
(65) 421, 1986.
333. Yamamoto,Y., Ohno,Y., “Analysis of heavy oils by FID-TLC. (part 3).
Preparation of thin layer rods with chemically bonded silica gels.” Sekiyu
Gakkaishi (28) 493, 1985.
334. Leazar,L.L., “Quantitative analysis of petroleum residues and heavy oils
by TLC/FID.” J. Chromatogr. Sci. (24) 340, 1986.
335. Selucky,M.L., “Quantitative class separation of coal liquids using thin-layer
chromatography with flame ionization detection.” Lipids (20) 546, 1985.
336. Yokoyama,S., Umematsu,J., Inoue,K., Katoh,T., Sanada,Y., “Estimation of
compound classes in coal hydrogenation liquids by thin-layer chromatography.”
Fuel (63) 984, 1984.
337. Sol,B., Romero,E., Carbognani,L., Sanchez,V., Sucre,L., “Estudio comparativo
entre HPLC Y TLC-FID para el analisis cuantitativo de fracciones pesadas de
petroleo.” Rev. Tec. Intevep. (4) 127, 1984.
338. Yamamoto,Y., “Class separation of higher hydrocarbons by thin-layer
chromatography with flame ionization detection.” Bull. Chem Soc. Jpn. (58)
2569, 1985.
339. Nakata,S., Yamamoto,S., Takahashi,H., Shiroto,Y., “Rapid trace analysis of
hydrocarbon types for heavy oils by TLC/FID using silica-rod combined with
FI-MS.” Sekiyu Gakkaishi (30) 438, 1988.
340. Holmes,S.A., “Comparisons of hydrocarbon and nitorogen distributions in
geologically diverse tar sand bitumen.” Prepr.-Am.Chem.Soc.,Div.Pet.Chem.
(33) 247, 1988.
341. Yamamoto,Y., “Analysis of heavy oils by thin-layer chromatography with
flame ionization detection.” Sekiyu Gakkaishi (31) 351, 1988.
342. Karlsen,D.A., Larter,S., “A rapid correlation method for petroleum population
mapping within individual petroleum reservoirs:applications to petroleum
reservoir description.” Norwegian Petroleum Society 77, 1989.
343. Holmes,S.A., “Sequential hydrocarbon and nitrogen detection in thin-layer
chromatographic analysis of oil shale bitumen.” J. Chromatogr. (465) 345,
1989.
344. Perry,M.B., Cillo,D.L., Barrage,T.C., “Characterization of coal reaction
products using preparative-scale liquid chromatography and automated
thin-layer chromatography.” Symposium on analytical chemistry of heavy
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oils/resids presented before the division of petroleum chemistry, Inc. American
chemical society dallas meeting, April 9-14, 1989.
345. Karlsen,D.A., Larter,S.R., “Analysis of petroleum fractions by TLC-FID:
applications to petroleum reservoir description.” Org. Geochem, 17(5) 603,
1991.
346. Rauhala,M., “Separation of bitumens into fractions using two different
methods and characterization of fractions.” Chemistry of Bitumens
Symposium - Rome, June, 1991.
347. Zwijsen,M., Goosse,S., Teugels,W., Vandeneede,H., “Analysis of the generic
composition of asphalt by the Iatroscan method.” Chemistry of Bitumens
Symposium - Rome, June, 1991.
348. Madella,A., Le Bourlot,F., Brule,B., “Selection of asphalt cements for asphalt
cements/eva blends.” Chemistry of Bitumens Symposium - Rome, June,
1991.
349. Thyrion,F., Cogneau,P., Zwijsen,M., “Method for the evaluation of bitumens
based on the characteristics of bitumen-polymer mixes.” Chemistry of
Bitumens Symposium - Rome, June, 1991.
350. Bredael,P., Andriolo,P., Killens,E., “A structural study of the hot storage
stability of SBS-modified bitumens.” Chemistry of Bitumens Symposium -
Rome, June, 1991.
351. Wan,C.C., Waters,T.H., Wolever,R.D., “Development of a reproducible
Iatroscan method to chemically characterize asphalt.” Amer. Chem. Soc.,
37(3) 1350, 1992.
352. Venkateswaran,K., Kanai,S., Tanaka,H., Miyachi,S., “Vertical distribution
and biodegradation activity of oil-degrading bacteria in the Pacific Ocean.” J
Mar Biotecnol (1) 33, 1993.
353. Friedbacher,E.C., Schindlbauer,H., “Analysis of bitumen by TLC-FID
compared with column chromatography.” Bitumen 55(4) 149, 1993.
354. Weber,J.V., Krzton,A., Cebolla,V., Swistek,M., “Chromatographic
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428. Ranny,M., Zbirousky,M., Konecny,V., “TLC-FID of metolachlor.” Journal
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429. Ayyangar,N.R., Biswas,S.S., Tambe,A.S., “Separation of opium alkaloids by
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430. Gromek,D., Kisiel,W., Stojakowska,A., Kohlmunzer,S., “Attempts of chemical
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432. Romero,A.J.R., Herrera,J.C., De Aparicil,E.M., Cuevas,E.A.M., “Thin-layer
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433. Orinak,A., Matisova,E., Gyoryova,K., Slesarova,L., “Chromatographic
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436. StojaKowska,A., “TLC/FID in quantificaion of proscillaridin A in Urginea
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44 / 188
APPENDIX Ⅱ
IATROSCAN Application Data
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IATROSCAN ANALYSIS
1A
Keys to Analytical Procedure
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1
IATROSCAN Analysis (1A ) Keys to Analytical Procedure
― Contents ―
１．Development Solvent ・・・・・・・・・・・・・・・・・・・・・・・・ 2－4
A） Preparation of development solvent
B） Volume of development solvent
C） Setting of development tank
D） Temperature
E） Atmosphere in the development tank to be saturated by solvent vapor
２．CHROMAROD ・・・・・・・・・・・・・・・・・・・・・・・・・ 4－6
A） Activation of CHROMAROD
B） Pretreatment of CHROMAROD for phosphorous compound analysis
C） Special treatment of CHROMAROD
３．Sample spotting ・・・・・・・・・・・・・・・・・・・・・・・・・・ 7－8
A） Solvent for dissolving sample
B） Spotting
C） Removal of solvent dissolving sample
４．Influence of humidity on the performance of CHROMAROD ・・・・・・ 8
５．Development ・・・・・・・・・・・・・・・・・・・・・・・・・・・ 9
６．Removal of development solvent ・・・・・・・・・・・・・・・・・・ 10
７．FID／FPD Detection ・・・・・・・・・・・・・・・・・・・・・・・ 10－11
A） Detection by means of FID（Flame Ionizing Detector）
B） Dual detection by means of FID／FPD（Flame Photometric Detector）
８．Washing and Storage of CHROMAROD ・・・・・・・・・・・・・・ 11－13
A） Washing of CHROMAROD
B） Storage of CHROMAROD
R
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2
IATROSCAN Analysis (1A ) Keys to Analytical Procedure
These “Keys to Analytical Procedure”are for your better IATROSCAN analysis.
1．Development Solvent
Ａ）Preparation of development solvent
Even a slight difference in composition of development solvent has a great influence on
chromatogram pattern. Therefore, preparation of development solvent shall be precisely
made with clean measuring cylinder and/or measuring pipette.
In addition, the development solvent shall be newly prepared everyday since the
composition of prepared development solvent will be gradually varied by its evaporation
as time goes on.
Ｂ）Volume of development solvent
To always keep constant the distance from the level of development solvent to the samplespotting point, the volumes of development solvent and tank shall always be constant.Needless to say, a clean development tank shall always be used. As for the volume of
development solvent, please refer the table below.
Development Tank Recommended Volume of Development Solvent
DT－150 70ｍL
Ｃ）Setting of development tank
Set the development tank on the level table at almost constant temperature, which shall
be kept from direct sunlight and air turbulence. If the temperature of whole tank is not
constant, stable separation of components in the sample can not be obtained because the
development tank can not be filled up by solvent vapor homogeneously.
R
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3
Ｄ）Temperature
Development work shall be executed always under constant temperature. Should there be
some difference of temperature in the course of development, separation pattern and
development time will be varied (see Analytical Example-1). Analyst should always
confirm if there is no difference between the temperature of development solvent and that
of circumstance where the tank is set. Because, sometimes heat generation will be caused
during preparation of mixed development solvent.
ａ）Development at 20℃ ｂ）Development at 30℃
Analytical Example-1: Separation pattern affected by development temperature
The above chromatograms are the examples of analyzing lipids at different temperature [(a)20℃,(b) 30℃], where phosphatidyl choline and phosphatidyl inositol are separated at 20℃, but overlappedeach other at 30℃.Sample： Standard Mixture
1.Cholesterol, 2.Phosphatidyl ethanolamine, 3.Phosphatidyl choline, 4.Phosphatidyl inositol,5.Sphingomyelin, 6.Lysophosphatidyl choline
Stationary Phase ：Chromarod-SⅢMobile Phase ：Chloroform:Methanol:Water:25%Ammonia 47:20:2.5:0.28Measurement conditions：Hydrogen 160mL/min, Air 1.5L/min, Scan Speed 40sec/scan
Ｅ）Saturating inside of development tank with solvent vapor
It is very important to always establish constant atmosphere inside the tank saturated
with solvent vapor in order to acquire good analytical result and reproducibility. To
accomplish this, therefore, perform the following procedures.
①Erect a new filter paper inside development tank and wet it with solvent.
・ DT－150：After erecting a L-shaped filter paper along one side of the tank, wet it with
solvent and then cover the tank with its lid
1
2
3 4
5
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2 3 4
5
6
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4
14cm
17cm
2.5cm
DT-150
②Leave the tank with filter paper and solvent as it is until it become saturated with
solvent vapor.
③Wet again the filter paper with solvent
As time passes, the solvent vapor filled inside the tank tends to move downward,
resulting in poor saturation in its upper portion even if the filter paper wetted with
solvent is in it. If the development were executed under above mentioned state, the
development time is possibly prolonged and the separation pattern vary. Accordingly,
make sure to wet again the filter paper with solvent in advance to development.
２．Chromarod
Chromarod, developed exclusively for IATROSCAN, is a thin layer in the form of thin
quartz rod evenly applied and sintered inorganic binder and adsorbent (silica gel or
alumina) on it. Usually, the cleaning and activation of Chromarods can be achieved by
Blank scan on IATROSCAN, which enables Chromarods to be used repeatedly.
However, their repeated use of Chromarod is possible in the limited case where thedevelopment solvent with same composition is used, because Chromarod has a memoryeffect to fix a bit acidic and/or alkaline substances on it. Therefore, the composition of
development solvent should be always same for repeated use of Chromarod.Especially, in case of analyzing a sample having radical(s) dissociation constant affectedby pH, good reproducibility will not be achieved as the case may be. For instance, the
separation pattern of the sample developed by neutral solvent on a new Chromarod isdifferent from that of the Chromarod treated with acidic development solvent.
The expected number of repeat use of Chromarod is approx. 20, which depends on how
the rod is used.
Chromarod is made of quartz glass. Handle it with thorough care.
Filter
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5
12345678910
100
50
100
50
Rod Holder
1)
Retaining Clip
2)
Ａ）Activation of Chromarod
Activation of Chromarod can be easily conducted only by Blank scan once to twice onIATROSCAN, the conditions of which shall be same as those of actual analysis (ex.hydrogen 160mL/min, air 2.0L/min, scan speed 30sec/scan). Activation and cleaning of
Chromarod can be performed at the same time.
The activation of Chromarod shall be performed immediately before sample spotting. And,
start development without delay right after spotting.
Organic compounds originated from the shipping package (cushion material and box) arebeing adsorbed on the Chromarods. These organic compounds are on the parts of both
ends of Chromarod as well, which the hydrogen flame can not reach.
Therefore, if the Chromarods are used as they are, these organic compounds will cause
the ghost peak in the course of measurement. In consideration of the above, remove the
adsorbed matters on the Chromarods in the shipping package in a following manner.
①Place the Chromarod(s) in position on the rod holder.
②Put the rod holder bearing Chromarod(s)
in a development tank containing the
same development solvent as that to be
used for actual analysis, and execute
development to the extent that the
solvent front crosses the sample spotting
point.
Retaining Clip
TLC Development Tank DT-150
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6
③Take out the set of rod holder and Chromatod(s), and vaporize the development solvent
on them [see 6. Removal of Development Solvent (page 10 )].
④Conduct Blank Scan twice in order to remove the organic compounds on the Chromarod(s),
and clean and activate the latter.
Ｂ）Treatment of Chromarod for analyzing phosphorous compound
Phosphorous component sometime remains on the rod even after IATROSCAN analysisdue to its interaction with Chromarod. Accordingly, the pretreatment and washing ofChromarod(s) is required for selective measurement of phosphorous by FPD of
IATROSCAN MK-6. For the details, refer the following.
・IATROSCAN Analysis (4A)「Chromarod-SⅢ Treatment Procedure for FPD Analyzing
Phosphorous Compound」
Ｃ）Preparation of Chromarod specially treated
Even special types of thin-layer treated with silver or cupper nitrate or boric acid are
required, they are easily obtained by easy pretreatment as shown in the followingreferences for further details.
・IATROSCAN Analysis (12) 「Analysis of glyceride isomers by Chromarod treated with
boric acid 」
・IATROSCAN Analysis (16) 「Analysis of molecular type of glycerides by Chromarod
treated with silver nitrate」
・IATROSCAN Analysis (22) 「Analysis of lipids by Chromarod-SⅢ treated with cupper
nitrate」
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３．Sample spotting
Minimizing the size of sample spot is a must for successful separation. The following
points should be kept in mind in the spotting procedure.
Ａ）Sample preparation
① Solvent for dissolving sample
Select a solvent of the lowest possible boiling point and polarity among those which can
sufficiently dissolve the sample. The spot tends to spread more when a solvent of higher
boiling point or polarity is used. Generally, hexane, benzene and chloroform are suitable
solvents.
②Concentration of sample solution
Prepare a sample solution with the concentration of approx. 2～5μg/peak. In case the
concentration of the sample concerned is unknown, adjust the concentration of sample to
1～2% (10～20mg/ｍL ). It is recommended to keep the sample solution prepared in a vial
with a screw-type of organic solvent-proof lid.
Ｂ）Sample spotting
Spot the sample solution on the rod(s) by means of micro-dispenser attached.The appropriate spotting size for better quantification is 1μl. Firstly, remove the set ofRod Holder and activated Chromarods from the scanning stage and place it onto the
attached Spotting Guide. Then, lift up a bit the upper part of Rod Holder so as to havethe lower ends of Chromarods touch the bottom of Rod Holder.Spot samples solution on the origin points of Chromarods (i.e. Zero point graduation of
Rod Holder) with the guide of white spotting line of Spotting Guide.
Micro-Dispenser
Spotting Line
(white color)
Spotting Guide
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Spotting shall be carefully conducted since analytical results will remarkably be affected
by spotting reproducibility. To make the spotted width as narrow as possible, eject the
sample bit by bit in several installments. Broad spotting width will result in worse
separation.
During spotting, if the tip of micro-dispenser does not contact the Chromarod, some part
of sample solution will climb up the outside of needle, which, as a result, makes it
impossible to spot whole sample solution sucked in on the Chromarod. To minimize
spotting error, therefore, eject the sample solution under the state of micro-dispenser tip
contacting Chromarod, which enables to transfer the weighed sample solution onto the
rods. And, in this case, do not pressurize the Chromarod since it is fragile and dangerous
in case of breakage.
In case that the spotting with are broaden due to the nonvolatile solvent of sample
solution, blow air (cool) upon the spotted area to accelerate its vaporization by hand dryer
in each interval of a bit spotting.
Ｃ）Removal of sample dissolving solvent
Upon spotting, remove the sample dissolving solvent on the rod for certain. Blowing the
rod by means of hand-dryer is recommended to remove the dissolving solvent with high
boiling point.
４．Relation between Chromarod performance and humidity
In case of using development solvent with relatively high polarity, there can be observed
little affection of humidity on the development pattern. But, the development is affected
in a large way by the solvent with low polarity. Therefore, the development work shall be
executed as quickly as possible upon completing sample spotting.
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５．Development
Put the Chromarods spotted the sample on them in the development tank saturated
enough with the vapor of development solvent (see Item 1., “Development Solvent”in the
page 3), and continue the development until the solvent front reaches the targeted
development distance.
In case of using the development tank DT-150, the adsorption of solvent to the
Chromarods is improved by leaning the rod holder against the filter paper in.
The development distance shall always kept
constant. Should the development distance
be varied, the counts of peak area measured
will be changed due to the transformation of
the peak shape(see the AnalyticalExample-2,
below).
When a multi-development is executed,
proceed with the next development upon
removing the solvent by letting the
Chromarods alone for 2 minutes at room
temperature or by blowing them by means
of had-dryer (cool wind).
c）7.5cm
Development Count ( Area )
Distance (cm)
a 2.5cm 9428
d）10cm b 5.0cm 7735
c 7.5cm 7597
d 10.0cm 6744
Analytical example-2: Relation of Peak Area with its Development Distance
The above data are the example of analyzing Cholesterol ester (2μg) corresponding to the differentdevelopment distance, where the peak width becomes broader and its height becomes lower inproportion to the development distance. The longer development distance is, the smaller the peak areacounts are.Stationary Phase： Chromarod-SⅢMobile Phase： Hexane:Diethyl ether 63:7（20℃）
Measuring Conditions：Hydrogen 160mL/min, Air 2.0L/min, Scan Speed 30sec/scan
b）5.0cm
a）2.5cm
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Rod Dryer TK-8
６．Removal of Development Solvent from Chromarod
Remove the development solvent from the Chromarods after completion of development.
If the measurement is started with the Chromarods still adsorbing the solvent on them, it
will cause the fluctuation of baseline and show the smaller peak area counts. The
remained solvent shall be entirely remove by keeping the Chromarods in the Rod Dryer
TK-8 exclusively used for Chromarods or in the appropriate drying oven, or by blowing
cool air against the Chromarods. However, it is enough to remove the solvent by air-dry
(for 2 minutes) at room temperature in case of volatile solvents like hexane and ether.
７．FID / FPD Measurement
The detection sensitivity of FID or FPD is varied by the flow rate of air and/or
hydrogen and scanning speed. Therefore, set the hydrogen flow rate to 160mL/min.
There is a possibility that some parts of separated components will remain unburned on
the rods under the low hydrogen flow rate even after scanning. On the other hand, under
the high hydrogen flow rate, the rods will unnecessarily be overheated and the
correlation between the peak area counts and concentration of separated component will
be damaged.
Ａ）FID ( Flame Ionization Detector ) Measurement [ Single detection ]
Following measurement conditions shall be applied for FID single measurement.
・Hydrogen 160mL／min，Air 2.0L／min，Scan Speed 30sec／scan
The peak shape and intensity of each component will be varied by the boiling point of
component, its interaction with the rod, scanning conditions and so on. Therefore, the
comparison of analytical data shall be made among those obtained under the same
scanning conditions.
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Ｂ）FID/FPD ( Flame Photometric Detector ) Measurement [ Dual detection ]
When analyzing sulfur (S) or phosphorous (P) by means of FPD, the FPD selectivity for
detecting S or P depends on the flow rates of hydrogen and air and scanning speed.
Therefore, the recommended conditions to selectively detect S or P are as follows;
・Selective detection of S by FPD
Interference filter：394nm
Hydrogen 160mL／min，Air 0.5L／min，Scan Speed 30sec／scan
・Selective detection of P by FPD
Interference filter：526nm
Hydrogen 160mL／min，Air 1.5L／min，Scan Speed 40sec／scan
8．Washing and storage of Chromarod(s)
Ａ）Washing of Chromarods
Usually, there is no need to wash Chromarods. The Chromarods can be used again for a
next sample measurement in succession to the previous measurement as the Chromarods
are cleaned and activated at the same time of measurement on the Iatroscan. The Blank
Scan on the Iatroscan is enough to clean the Chromarods.
The expected number of repeated use of Chromarod is approx. 20 times. However, in case
of selective measurement of P by FPD of Iatroscan MK-6, the pretreatment and washing
of Chromarods are required.
For further information, refer the Iatroscan Analytical Method titled 「 4A : Chromarod-
SⅢ Treatment Procedure for FPD Analyzing Phosphorous Compound」.
・IATROSCAN Analysis (4A)「Chromarod-SⅢ Treatment Procedure for FPD Analyzing
Phosphorous Compound」
The phosphorous or metallic component(s) remained on the Chromarod(s) can not be
removed by the scanning only. Therefore, the cleaning procedure below should be followed
to eliminate those residues. Should there still be some noise on the chromatograph even
after such cleaning procedure, replace the Chromarod(s) with new one(s).
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①Removal of P compound(s)
Following procedure is for removing P compound(s) by using ammonia water. However,
note that the separation pattern to be obtained after the cleaning will sometimes be
different from that of previous measurement as the development conditions may be,
because the ammonia used will possibly remain on the rod(s) even after finishing this
cleaning.
In addition, during this cleaning, the silica gel as adsorbent of Chromarod-SⅢ will be
gradually dissolved due to its poor alkali resistance. Accordingly, the Chromarods shall
not be soaked in ammonia solution for long time, which will cause inferior separation due
to silica gel degradation.
1）Immerse the Chromarods in the 2.5% ammonia solution for 10 minutes.
2）After taking out the Chromarods from the 2.5% ammonia solution, thoroughly rinse
them with deionized water.
3）Remove the water by drying the Chromarods for one (1) hour at 120℃ , or by
conducting Blank scan twice after drying them for three (3) minutes at 120℃.
②Removal of metallic salt(s)
As for the metallic salts such as NaCl and CaCl2 which can not be removed by scanning,
remove eliminate them in accordance with the following procedure. Nevertheless, note
that the separation pattern to be obtained after the cleaning will sometimes be different
from that of previous measurement as the development conditions may be
1）Wash the Chromarods lightly with deionized water.
2）Soak them in the concentrated nitric acid all the night through.
3）Upon taking out the Chromarods from the concentrated nitric acid , thoroughly rinse
them with deionized water.
4）Remove the water by drying the Chromarods for one (1) hour at 120℃ , or by
conducting Blank scan twice after drying them for three (3) minutes at 120℃.
③Removal of organic compound(s)
Most organic compounds can be eliminated by the Blank Scan. Execute the Origin Scan
several times to remove the organic compound(s) remained on the origin point. In case
that the organic compounds are remaining on the whole of Chromarod, conduct the Blank
Scan twice at the scanning speed of 60sec/scan.
If the organic residuals can not be removed by the scanning, wash the Chromarods with
concentrated sulfuric acid in the manner described below. However, note that the
separation pattern to be obtained after the cleaning will sometimes be different from that
of previous measurement as the development conditions may be, because the sulfuric acid
used will possibly remain on the rod(s) even after finishing this cleaning.
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1）Wash the Chromarods lightly with deionized water.
2）Soak them in the concentrated sulfuric acid all the night through.
3）Upon taking out the Chromarods from the concentrated sulfuric acid, thoroughly
rinse them with deionized water.
4）Remove the water by drying the Chromarods for one (1) hour at 120℃ , or by
conducting Blank scan twice after drying them for three (3) minutes at 120℃.
B）Storage of Chromarods
Keep the Chromarods in a clean glass vessel with a lid to avoid their contamination
caused by adsorption of organic/inorganic matter coming from the dust in the air.
Never storage them in the vessel containing drying agent. MITSUBISHI CHEMICAL
MEDIENCE CORPORATION is ready to supply the Rod Storage Chamber DE-3 as an
optional item.
Rod Strage Chamber DE-3
Do not expose Chromarods to high humidity and/or water, which
damages their thin layers.
Products Guide：
Code No. Product Name
3248 Chromarod-SⅢ（silica type） 10pcs/box
3201 Development Tank DT-150
3215 Pre-cut Filter Papers for DT-150 Development Tank 100pcs
3221 Rod Storage Chamber DE-3
5101 Rod Dryer TK-8
6321 Rod Holder SD-6
6100 Microdispenser
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IATROSCAN ANALYSIS
2A
Type Analysis for Heavy Oil
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IATROSCAN Analysis (2A) Type analysis for Heavy oil
１．Heavy oil type analysis by Iatroscan
Heavy oil is composed of a various complex mixture. The heavy oil analysis is done by the
whole characterization (such as elementary analysis or NMR), the type analysis and so forth.
The type analysis by Iatroscan is one of composite analysis. The heavy oil is separated into 4
groups (saturated hydrocarbons, aromatic hydrocarbons, resins, asphaltenes) by multiple
development using 3 kind of development solutions different polarity. Compare with the type
analysis by liquid chromatography, this analysis is short time/easy to handle.
２．An example of the heavy oil analysis by Iatroscan
Ａ）The method of the heavy oil analysis
Sample ： Heavy oil1) 10mg／mL (dissolved by dichloromethane)
and 5，10，20，30mg／ｍL (for response curve)
Spotting Volume ： 1μL (using the Micro-dispenser : Code No.6100)
Stationary phase ： Chromarod -SⅢ（Code No.3248）
Development tank ： DT-150（Code No.3201）
Mobile phase2) ： 1st. Hexane 100%（60mL） 10 cm（20℃）
2nd. Toluene 100%（60mL） 6 cm（20℃）
3rd. Dichloromethane:Methanol 57:3 2.5cm（20℃）
（After each development, proceed with the next development after
removing the development solvent from the Chromarods by air-dry for
2 minutes at room temperature.）
Measurement conditions： Hydrogen 160mL／min, Air 2.0L／min, Scan Speed 30sec／scan
Detector ： FID
First, conduct Blank Scan （Hydrogen 160mL／min, Air 2.0L／min, Scan Speed 30sec／scan）
in order to clean and activate the Chromarods. １μL of the sample solution 10mg／mL was
spotted3) on the Chromarod by means of the micro-dispenser. Upon spotting, remove the
sample dissolving solvent on the Chromarods by air-dry for 2 minutes at room temperature.
１） Before sampling a heavy oil, be sure to mix it very well because these components of theheavy oil are broken up equally. It is recommended to keep the sample solution prepared in a vialwith a screw-type of organic solvent-proof lid.
２） The grade of organic solvents should be liquid-chromatography or more pure than it.
３ ） To make the spotted width as narrow as possible, eject the sample bit by bit in severalinstallments(8-10times/1μL). To minimize spotting error, eject the sample solution under the state ofmicro-dispenser tip contacting Chromarod, which enables to transfer the weighed sample solution ontothe rods. And, in this case, do not pressurize the Chromarod since it is fragile and dangerous in case ofbreakage.
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Put the Chromarods spotted the sample on them in the first development tank (Hexane)
saturated enough with the vapor of the development solvent, and continue the development
until the solvent front reaches 100-point graduation of the Rod Holder. After the development,
remove the development solvent on the Chromarods by air-dry for 2 minutes at room
temperature. Next, the Chromarods were developed in Toluene until the solvent front
reaches 60-point graduation of the Rod Holder, and dried at room temperature for 2 minutes.
In the third stage of development, the Chromarods were developed in
Dichloromethane:Methanol(57:3) until the solvent front reaches 25-point graduation of the
Rod Holder.
The Chromarods were then dried at room temperature for 2 minutes and measured4) by FID
（Hydrogen 160mL／min, Air 2.0L／min, Scan Speed 30sec／scan）.
The heavy oil sample was measured using ten (10) Chromarods and calculated each
percentage of 4 groups (saturated hydrocarbons, aromatic hydrocarbons, resins,
asphaltenes) of the heavy oil. The chromatogram is shown in Figure-1. The data and the
reproducibility of daily error are shown in Table-１and Figure-2 respectively.
Also the response curve s for the heavy oil (5～30μg) are shown in Figure-3.
1
2
3
4
Figure-1）Chromatogram of heavy oilSample： Heavy oil
1.Saturated hydrocarbons, 2.Aromatic hydrocarbons, 3.Resins, 4.AsphaltenesStationary Phase：Chromarod-SⅢMobile Phase： 1st, Hexane 100% 10cm
2nd, Toluene 100% 6cm3rd, Dichloromethane:Methanol 57:3 2.5cm
Measurement conditions：Hydrogen 160mL/min, Air 2.0L/min, Scan Speed 30sec/scan
４）Asphaltenes are not eluted and are hence susceptible to the problem of rod rotation which influencesresults. Rotate the rod and the asphaltenes that remain at the point of application areoriented toward the burner with respect to the detector for valid quantification.
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Table-1）Measured values of 10 Chromarods
ChromarodNo.
Peak 1Saturated HC
Peak 2Aromatic HC
Peak 3Resins
Peak 4Asphaltenes
123456789
10
38.435.037.636.735.136.036.534.937.038.2
43.745.343.843.645.044.244.544.543.342.7
12.113.312.013.613.313.613.314.312.812.8
5.76.46.66.06.66.25.86.46.96.4
XSDCV（％）
36.51.33.5
44.10.81.8
13.10.75.4
6.30.45.7
Figure-2）Reproducibility of daily error
（％）
50
40
30
1 2 3 4 5 （day）
1．Saturated HC （％）
50
40
30
1 2 3 4 5 （day）
2．Aromatic HC
（％）
20
10
0
1 2 3 4 5 （day）
3．Resins （％）
20
10
0
1 2 3 4 5 （day）
4．Asphaltenes
% % % %
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芳香分
飽和分
レジン
アスファルテン
Figure-3）Response curves for heavy oil
Prepared heavy oil sample solutions（5,10,20,30ｍｇ/ｍL）and measured with Iatroscan by the aboveconditions. The response curves of each group were found to be linearly proportional to the amount ofheavy oil.
0
20000
40000
60000
80000
100000
120000
140000
0 10 20 30 40常圧残油 （μｇ）
ピーク面
積（c
ounts）
Saturate HC
Aromatic HC
Resins
Asphaltenes
Pea
karea
(coun
ts)
Heavy oil (μg )
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11-Ⅱ
Analysis of Biodegraded Crude Oil by IATROSCAN
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IATROSCAN ANALYSIS
5
Analysis of Lipid by Iatroscan
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IATROSCAN ANALYSIS
5-Ⅱ
Analysis of Lipid by Iatroscan
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IATROSCAN ANALYSIS
23
Analysis of Lipid by Iatroscan (Marine Products)
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IATROSCAN ANALYSIS
4A
Chromarod-SⅢ Treatment Procedure for FPD Analyzing PhosphorousCompound
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IATROSCAN Analysis (4A)
Chromarod-SⅢ Treatment Procedure for FPD Analyzing Phosphorous Compound
The special treatments (pretreatment・before-analysis treatment・after-analysis washing)
for Chromarod (s) are required for selective measurement of phosphorous by FPD (Flame
Photometric Detector) of IATROSCAN MK-6.
The pretreatment of Chromarod-SⅢ is necessary for keeping the base stability in case of
selective measurement of phosphorous by FPD. The before-analysis treatment is necessary
for the analytical reproducibility. The after-analysis washing is necessary for removing the
remained P compound(s) from the Chromarod. In case of the selective measurement of P by
FPD, be sure to do these treatments.
1．Pretreatment procedure of the Chromarod (When analyzing P by means of FPD)
To begin with, the new Chromarod(s) is treated in a following manner. By means of the
pre-treatment, the surface of the Chromarod is washed and the base in case of selective
measurement of P by FPD becomes stable.
Pretreatment procedure for the Chromarod
Chromarod-SⅢ (new)
↓
steep the new Chromarod in 6N HCl solution for 22-24hour
↓
wash the rod with distilled water for 1 minute (repeat twice)
↓
dry it at 120℃ for 1hour
or
dry 3 minutes at 120℃ and conduct Blank Scan twice.
(Hydrogen 160mL/min, Air 1.5L/min, Scan Speed 40sec/scan)
↓
［go to the before-analysis treatment（or storage）］
（In case the pretreated Chromarod is stored for future use, keep the Chromarod in a
clean vessel with a lid.）
Caution: Chromarod is fragile. Handle with care.
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２．Before-analysis treatment procedure of the Chromarod
In case of analyzing a sample having radical(s) with dissociation constant affected by pH,
such as phospholipid, there are some cases where the separation pattern becomes
stable by using acidic or alkaline development solvent. If a sample development is carried
out by use of a development solvent containing acid or alkaline, the pH on the Chromarod is
memorized. In case of multi-development, the development should be performed by solvents
such as a combination of alkaline and alkaline or a combination of alkaline and neutral.
When amphoteric-electrolytes are developed by using a combination of acidic and alkaline
solvents, the separation pattern may be unstable because the pH on the Chromarod can not
be controlled.
The before-analysis treatment for Chromarod is shown on a following manner. MITSUBISHI
CHEMICAL MEDIENCE CORPORATION recommends the following treatment methods
when phosphorus compounds are analyzed by using alkaline development solvent.
Before-analysis treatment
The pre-treated Chromarod-SⅢ
↓
Blank Scan twice (Hydrogen 160mL/min, Air 1.5L/min, Scan Speed 40sec/scan)
↓
soak it in 2.5％ NH4OH solution for 10 minutes.
↓
wash the rod in distilled water for 1 minute (repeat twice)
↓
dry 3 minutes at 120℃ and the Blank Scan twice
↓
［for analyzing］
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３．After analysis treatment procedure
Phosphorous component sometimes remains on the rod even after IATROSCAN analysis due
to its interaction with Chromarod. After the analysis, the Chromarod is required to wash in a
following manner. Most of the P compound(s) can be eliminated by the washing procedure.
The adsorbent of Chromarod-SⅢ is silica gel. The silica gel is gradually dissolved due to its
poor alkali resistance during this washing. Therefore the expected number of repeat use of the
Chromarod is approx. 7 although it depends on how the rod is used.
From the reproducibility point of view, the Chromarod used for the FPD(P) measurement is
valid only in a day. If the Chromarod is used the next day, the separation pattern will be
different from that of the previous day.
After-analysis washing
The used Chromarod-SⅢ
↓
soak it in 2.5％ NH4OH solution for 10 minutes.
↓
wash the rod in distilled water for 1 minute (repeat twice)
↓
dry 3 minutes at 120℃ and the Blank Scan twice
↓
［for analyzing］
Caution:
Do not use treated Chromarod with the special treatments for any analysis
other than the phosphorus analysis, otherwise a good reproducibility will
not be obtained by unstable pH on the Chromarod.
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４．Analysis of phospholipids（examples of applications）
Ａ）Application of lipids (standards mixture) analysis using the alkaline development solvent.
Sample ： 1.Cholesterol(Nu-Chek-Prep.), 2.Phosphatidyl ethanolamine(egg:Avanti-
Polar-Lipids), 3.Phosphatidyl choline(Wako Chemical), 4.Sphingomyelin
(Avanti-Polar-Lipids)
Mobile Phase：Chloroform:Methanol:DW:25%Ammonia = 47:20:2.5:0.28 (20℃)
Detectors： FID (Flame ionization detector) and FPD
Interference Filter：The Interference Filter for Phosphorous (526nm)
Iatroscan MK-6 conditions：H2 160mL/min，Air 1.5L/min，Scan Speed 40sec/scan
Preparing a sample solution, the four lipids（Cholesterol／Phosphatidyl ethanolamine／
Phosphatidyl choline／Sphingomyelin：@3mg/mL）were dissolved by Chloroform:MeOH(2:1).
1μL of the sample solution was spotted on the special treated Chromarod-SⅢ by means of
micro- dispenser. It was developed by Chloroform:Methanol:DW:25%Ammonia (47:20:2.5:0.28,
20℃) and measured by FID and FPD(P) simultaneously.
1
23
4FID
FPD(P)
Figure-1) Lipids analysis
Sample： Standards Mixture1.Cholesterol, 2.Phosphatidyl ethanolamine, 3.Phosphatidyl choline, 4. Sphingomyelin
Stationary Phase： Chromarod-SⅢ（treated）Mobile Phase： Chloroform:Methanol:Water:25%Ammonia 47:20:2.5:0.28 (20℃)Measurement conditions：Hydrogen 160mL/min, Air 1.5L/min, Scan Speed 40sec/scan
Dual detections of FID and FPD(P)
The result is shown in figure 1. Every lipid in the sample was detected by FID, and the
phospholipids were detected by FPD(P) selectively.
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Ｂ）Application of lipids (standards mixture) analysis using the alkaline and neutral
development solvents.
Sample：1.Cholesterol palmitate(Tokyo Kasei), 2.Triglyceride(P-L Biochemicals) 3.Cholesterol,
4.Phosphatidyl ethanolamine, 5.Phosphatidyl choline, 6.Phosphatidyl inositol
(Avanti－Polar－Lipids), 7.Sphingomyelin, 8.Lysophosphatidyl choline
(Avanti-Polar-Lipids)
Mobile Phase：
1st, Chloroform:Methanol:DW:25%Ammonia = 47:20:2.5:0.28（20℃） 7cm
2nd, n-Hexane:Diethyl ether = 63:7（20℃） 10cm
Iatroscan MK-6 conditions：H2 160mL/min, Air 1.5L/min, Scan Speed 40s/Scan
1
2
3
4
5 6
7
8
FID
FPD(P)
Figure-2) Lipids analysis
Sample： Standard Mixture1.Cholesterol palmitate, 2.Triglyceride, 3. Cholesterol, 4.Phosphatidyl ethanolamine,5.Phosphatidyl choline, 6. Phosphatidyl inositol, 7. Sphingomyelin, 8.Lysophosphatidyl choline
Stationary Phase： Chromarod-SⅢ（treated）Mobile Phase： 1st, Chloroform:Methanol:Water:25%Ammonia 47:20:2.5:0.28 (20℃)
2nd, n-Hexane:Diethyl ether 63:7 (20℃)Measurement conditions：Hydrogen 160mL/min, Air 1.5L/min, Scan Speed 40sec/scan
Dual detections of FID and FPD(P)
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IATROSCAN ANALYSIS
12
Analysis of Glyceride isomers using Boric acid-Impregnated Chromarods
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IATROSCAN ANALYSIS
16
Analysis of Triglyceride Molecular Species using Silvernitrate-Impregnated Chromarods
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IATROSCAN ANALYSIS
22
Lipid Analysis using Copper Sulphate-Impregnated Chromarods
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IATROSCAN ANALYSIS
20
Experimental Analysis for Infinitesimal Components Contained in MainIngredients
(Detection of Infinitesimal Phospholipid in Edible Oils)
Infinitesimal amounts of impurity and additive contained in the main ingredients
may raise difficulties to analyze sometime. The data collected in this report
describes applications by combined with the Iatroscan and Column
chromatography to be used for detection of infinitesimal components contained in
Edible oils.
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13
Tracing of Reaction with Enzymatic Experimental Reaction by Iatroscan
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IATROSCAN ANALYSIS
9
Separation of Isomers and Derivatives
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IATROSCAN ANALYSIS
21
Analysis of Surface Active Agents by Iatroscan
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IATROSCAN ANALYSIS
24
Analysis of Polymer Additives by Iatroscan
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26
Environment and Health Friendly Developing Solvents for Iatroscan
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Dye (Food dye, Naphthol quinone, Azo dye)
Hormons (Pregnandiol)
Ginseng Saponin
Liquid Crystal
Capsaicine
Cosmetic Cream
Rubber Antioxidant
Polymer
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