Abstract
An undoubted advantage of TLC is the simplicity of qualifying a separation. Coloured compounds can be seen by eye. Compounds absorbing only below 400 nm can be visualized by incorporating a fluorescent dye in the layer. Illuminating the plate with light of 256 nm will induce fluorescence. A sample zone covers the dye and inhibits fluorescence, so the zone is visible as a dark band. Illuminating the plate with light of 366 nm can identify substances with native fluorescence. Under these conditions you can observe most of the fluorescent substances by eye.
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Jork H, Funk W, Fischer W, Wimmer H (1990, 1994) Thin Layer Chromatography, Volume 1a, Physical and chemical detection methods, Volume 1b, Reagents and detection methods. VCH Verlagsgesellschaft mbH, Weinheim
Dyeing reagents for thin layer and paper chromatography. Merck-Company. http://www.cob.uha.fr/site%20cot/toolbox.php_files/TLC%20bible.pdf
Krebs KG, Heusser D, Wimmer H (1969) In: Stahl E (ed) Thin-layer chromatography, a laboratory handbook. Springer, Berlin, pp 854–909
Touchstone JC, Dobbins MF (1978) Practise of thin layer chromatography. Wiley, New York, pp 135–187
Touchstone JC (1992) Practice of thin layer chromatography. Wiley, New York, pp 129–183
Kraus Lj, Koch A, Hoffstetter-Kuhn S (1995) Dünnschichtchromatographie. Springer, Berlin
Cimpan G (2001) In: Nyiredy Sz (ed) Planar chromatography, a restrospective view for the third millennium. Springer, Budapest, pp 410–445
Randerat K (1965) Dünnschicht-Chromatographie. Chemie, Weinheim
Wagner H, Bladt S (1996) Plant drug analysis. A thin layer chromatography atlas, 2nd edn. Springer, Berlin
Reimann K (1972) Schnellnachweis von Schwermetallionen mittels Dünnschicht-chromatographie der Dithizonate aus Wasser, Abwasser und Schlamm. Wasser-und Abwasser-Forschung 1:3–7
Baranowska I, Poreba R, Baranowski J, Aleksandrowicz R (1992) Separation and identification of metals in the human placenta by TLC. J Planar Chromatogr 5:469–471
Bruno P, Caselli M, Fracassi F, Traini A (1984) HPTLC separation and densitometric determination of some metallic dithizonates at subnanogram levels: applications to real samples. Anal Lett 17:397–412
Baranowska I, Baranowski J, Norska-Borówka I, Pieszko C (1996) Separation and identification of metals in human bones, placenta and milk and in air by absorption and ion-exchange thin-layer chromatography. J Chromatogr A 725:199–202
B. Spangenberg Unpublished results
König K-H, Kessler I, Schuster M, Steinbrech B (1985) Zur Chromatographie von Metallen. XV. Dünnschicht-Chromatographie von Rh, Pd, Ir und Tt mit 1-Hydroxy-2-pyridinthion. Fresenius Z Anal Chem 322:33–35
Steinbrecher B, König K-H (1983) Zur Chromatographie von Metallchelaten. XII. Dünnschicht-Chromatographie (DC) und Hochdruck-Flüssigkeits-Chromatogrphie (HPLC) der Metallchelate des 1-Hydroxy-2-pyridinthin (HPT). Fresenius Z Anal Chem 316:685–688
König K-H, Kessler I, Schuster M, Steinbrech B (1984) Zur Chromatographie von Metallchelaten. XV. Dünnschichtchromatographie von Rh, Pd, Ir und Pt mit 1-Hydroxy-2-pyridinthin. Fresenius Z Anal Chem 319:66–69
Brozovic-Pohl K, Altorfer A, Perlia X (1992) A new (limit) test for heavy metals. Fresenius Z Anal Chem 343:348–351
Siembida R, Rozylo TK, Jamrozek-Manko A (1998) Aspects of the exact quantitative determination of formaldehyde in tooth tissue by TLC. J Planar Chromatogr 11:417–420
Funk W (1994) Derivatisierungsreaktionen im Bereich der quantitativen Dünnschicht-Chromatographie. Fresenius Z Anal Chem 318:206–219
Moosbach E, Rosenboom J, Hartkamp H (1999) Thin-layer detector tubes – a new system for the determination of trace gases in air by gas/solid reactions followed by remission photometric determination. Gefahrstoffe – Reinhalt Luft 59:71–78
Kim SS, Gallaher DD, Csallany AS (1999) Lipophilic aldehydes and related carbonyl compounds in rat and human urine. Lipids 34:489–496
Büldt A, Karst U (1999) N-Methyl-4-hydrazino-7-nitrobenzofurazan as a new reagent for air monitoring of aldehydes and ketones. Anal Chem 71:1893–1898
B. Spangenberg Unpublished results
Gübitz G, Wintersteiger R, Frei RW (1984) Fluorigenic labelling of carbonyl compounds with 7-hydrazino-4-nitrobenzo-2-oxa-1,3-diazole (NBD-H). J Liq Chromatogr 7:839–854
Jork H, Funk W, Fischer W, Wimmer H (1988) In situ pre-chromatographic derivatization. J Planar Chromatogr 1:280–292
Miller JM, Kirchner JG (1953) Chromatostrips for identifying constituents of essential oils. Anal Chem 25:1107–1109
Kofoed J, Korczak-Fabierkiewicz C, Lucas GHW (1966) A study of the conversion of phenothiazine derivates to the corresponding sulfoxides on thin-layer plates. J Chromatogr 23:410–416
Wilk M, Hoppe U, Taupp W, Rochlitz J (1967) Reaktions-Chromatographie, ein Identi-fizierungsverfahren für Aromaten bei der Papier- und Dünnschichtchromatographie. J Chromatogr 27:311–316
Wilk M, Bez W, Rochlitz J (1966) Neue Reaktionen der carcinogenen Kohlenwasserstoffe 3,4-Benzpyren, 9,10-Dimethyl-1,2-Benzanthracen und 20-Methylcholanthren. Tetrahedron 22:2599–2608
Baggiolini M, Dewald B (1967) Kontrollierte Hydrolyse auf Kieselgel-Platten mit chromatographischer Trennung der Spaltprodukte. J Chromatogr 30:256–259
Elgamal MHA, Fayez MBE (1967) A new application of thin-layer chromatography. Chemical reactions induced on the adsorbent. Fresenius Z Anal Chem 226:408–417
Purdy SJ, Truter EV (1963) Constitution of the surface lipid from the leaves of Brassica oleracea. Isolation and quantitative fractionation. Proc Royal Soc (London), Ser B 158:536–543
Young-Duck Ha, Bergner K-G (1981) Dünnschichtchromatographische Untersuchung von Phenylharnstoff- und N-Phenylcarbamat-Rückständen. Dtsch Lebensm Rundschau 77:102–106
Krüger D, Wichtl M (1985) Digitalisglykoside. Identifizierung und Strukturaufklärung mittels Reaktions-Dünnschichtchromatographie. Dtsch Apoth Ztg 125:55–57
Schütz C, Schütz H (1973) Unterscheidung ungesättigter Barbiturate mittels Reaktionschromatographie. Dtsch Apoth Zrg 113:1559–1562
Polesuk J, Ma TS (1971) Organic synthesis on microgram scale. VIII. Chlorination of acetanilide and related compounds by means of molecular chlorine. Mikrochim Acta 662–666
Gänshirt H (1963) Nachweisverfahren für dünnschichtchromatographisch getrennte Wirkstoffe von Antineuralgie- und Schlaftabletten. Arch Pharmaz 296:73–79
Cargill DI (1962) The separation of cholesterol from related stanols and stanones by thin-layer chromatography. Analyst 87:865–869
Schütz C, Schütz H (1973) Unterscheidung von 3,5-Dioxo-1,2-diphenyl-4-n-butyl-pyrazolidin (Phenylbutazon) und 3,5-Dioxo-1,2-diphenyl-4-prenyl-pyrazolidin (Prenazon) mit Hilfe der Reaktionschromatographie. Arzneim Forsch 23:428–431
Wintersteiger R (1982) Determination of morphine and morphine-6-nicotinate by an in situ reaction on chromatograhic plates with dansyl chloride. Analyst 107:459–461
Gertz Chr, Böschemeyer L (1980) Verfahren zur Bestimmung verschiedener Mykotoxine in Lebensmitteln. Z Lebensm Unters Forsch 171:335–340
Goliński F, Grabarkiewics-Szczesna J (1984) Chemical confirmatory tests for ochratoxin A, citrin, penicillic acid, sterigmatocystin, and zearalenone performed directly on thin layer chromatographic plates. J Assoc Off Anal Chem 67:1108–1110
Frei RW, Lawrence JF (1973) Fluorigenic labelling in high-speed liquid chromatography. J Chromatogr 83:321–330
Scholten AHMT, van Buren C, Lawrence JF, Brinkman UATh, Frei RW (1979) A residue technique for urea herbicides using catalytic hydrolysis on silica gel plates. J Liq Chromatogr 2:607–617
Wintersteiger R (1988) Fluoreszenz-Derivatisierung in DC und HPLC. GIT 32:5–11
Marcus BJ, Ma TS (1968) Organic synthesis on the microgram scale II. Mikrochim Acta 436–441
Spangenberg B (2006) Fluorescence spectroscopy in planar chromatography. International symposium for thin layer chromatography, Berlin, October 2006
Müller D, Morlock G (2006) New validated method for determination of acrylamide on potato chips by planar chromatography. International symposium for thin layer chromatography, Berlin, October 2006
Gübitz G, Wintersteiger R (1980) Identification of drugs of abuse by high-performance thin-layer chromatography. J Anal Toxicol 4:141–143
Junker-Buchheit A, Jork H (1988) Prächromatographische in situ-Derivatisierung von Fettsäuren im Picomol-Bereich. Teil I: HPTLC fluoreszierender Monodansylpiperzin- und cadaverin-Derivate. Fresenius Z Anal Chem 331:387–393
Lankmayr EP, Budna KW, Müller K (1979) Bestimmung von Thiolverbindungen durch selektive Fluorescenzderivatisierung und HPLC. Fresenius Z Anal Chem 295:371–374
Lankmayer EP, Budna KW, Müller K, Nachtmann F, Rainer F (1981) Determination of D-penicillamine in serum by fluorescence derivatization and liquid column chromatography. J Chromatogr 222:249–255
Wintersteiger R, Gübitz G, Hartinger A (1980) Derivatisation of sympathomimetics with primary amino groups on thin-layer plates by spotting the sample spots with fluorescamine. Chromatographia 13:291–294
Pataki G, Borko J, Curtis HCh, Tancredi F (1968) Some recent advances in thin-layer chromatography. New applications in aminoacid peptide and nucleic acid chemistry. Chromatographia 1:406–417
Minyard JP, Tumlinson JH, Thompson AC, Hedin PA (1967) Gas–liquid thin-layer chromatographic derivatisation technique for the identification of alcohols. J Chromatogr 29:88–93
Junker-Buchheit A, Jork H (1989) Monodansyl cadaverine as a fluorescent marker for carboxylic acids – in situ prechromatographic derivatization. J Planar Chromatogr 2:65–70
Wintersteiger R, Wenninger-Weinzierl G (1981) Neues, empfindliches Verfahren zur dünnschicht-chromatographischen Analyse von Substanzen mit alkoholischer Hydroxylgruppe durch Reaktion mit Naphthylisocyanat. Fresenius Z Anal Chem 309:201–208
Stahl E (ed) (1969) Thin-layer chromatography, a laboratory handbook. Springer, Berlin, pp 854–909
Postaire E, Sarbach Ch, Delvordre P, Regnault C (1990) A new method of derivatisation in planar chromatography: overpressure derivatisation. J Planar Chromatogr 3:247–250
Reich E, Schibli A (2006) High-performance thin-layer chromatography for the analysis of medicinal plants. Thieme, New York
Spangenberg B, Lorenz K, Nasterlack St (2003) Fluorescence enhancement of pyrene measured by thin-layer chromatography with diode-array detection. J Planar Chromatogr 16:328-334
Jaryj E, Lorenz K, Spangenberg B (2008) A simple method for the quantification of urethane in spirits. J Liq Chromatogr Rel Technol 31:1–9
Wall PE (1997) Argentation HPTLC as an effective separation technique for cis/trans isomers of capsaicin. J Planar Chromatogr 10:4–9
Neu R (1954) Tetraphenyl-diboroxyd aus Natriumtetraphenylborat. Chem Ber 87:802–805
Göcer M, Hoferer K, Zipfel J, Spangenberg B (2009) A new method for the quantification of paraquat, diquat, difenzoquat, mepiquat and chloromequat in water by thin-layer chromatography. J Planar Chromatogr 22:59–63
Hiegel K, Spangenberg B (2009) A new method for the quantification of dequalinium-cations in pharmaceutical samples by absorption and fluorescence diode array thin-layer chromatography. J Chromatogr A 1216:5052–5056
Machata G, Schütz H, Bäumler J, Daldrup T, Gibitz H-J (1989) Empfehlungen zur klinisch-toxikologischen Analytik. Folge 5: Empfehlungen zur Dünnschichtchromatographie. VCH, Weinheim
Becker G, Eichler D, Nolting H-G, Thier H-P (1987) Dünnschichtchromatographie in der Rückstandsanalytik von Pflanzenschutzmitteln und deren Metaboliten. VCH, Weinheim
Broszat M, Brämer R, Spangenberg B (2008) A new method for quantification of melamine in milk by absorption diode-array chromatography. J Planar Chromatogr 21:469–470
Cimpan G (2001) In: Nyiredy Sz (ed) Planar chromatography. A retrospective view for the third millennium. Springer, Budapest, pp 410–445
Mohammad A (1997) Thin-layer chromatographic methods for the identification, estimation and separation of toxic metals in environmental samples. J Planar Chromatogr 10:48–54
Mohammad A, Ajmal M, Anwar S, Iraqi E (1996) Twenty-two years report on the thin-layer chromatography of inorganic mixtures: observations and future prospects. J Planar Chromatogr 9:318–360
Ahmad J (1996) Use of alumina stationary phase for thin layer chromatography of inorganic and organometallic compounds. J Planar Chromatogr 9:236–246
Spangenberg B Unpublished results
Segura R, Gotto AM (1974) A new fluorimetric procedure for the detection and quantitation of organic compounds in thin-layer chromatography. J Chromatogr 99:643–657
Hayakawa T, Hirati M (2003) An assay of ganglioside using fluorescence image analysis on a thin-layer chromatography plate. Anal Chem 75:6728–6731
Klaus R, Fischer W, Hauck HE (1989) Use of a new adsorbent in the separation and detection of glucose and fructose by HPTLC. Chromatographia 28:364–366
Klaus R, Fischer W, Hauck HE (1990) Application of a thermal in situ reaction for fluorimetric detection of carbohydrates on NH2 layers. Chromatographia 29:467–472
Klaus R, Fischer W, Hauck HE (1991) Qualitative and quantitative analysis of uric acid, creatine and creatinine together with carbohydrates in biological material. Chromatographia 32:307–316
Klaus R, Fischer W, Hauck HE (1993) Extension of the application of the thermal in-situ reaction on NH2 layers to the detection of catecholamines in biological materials. Chromatographia 37:133–143
Klaus R, Fischer W, Hauck HE (1994) Analysis and chromatographic separation of some steroid hormones on NH2 layers. Chromatographia 39:97–102
Stroka J, Arranz I, Mac Court J, Spangenberg B (2003) Simple and reliable working HPTLC quantification method for Sucralose®. J Liq Chromatogr Rel Technol 26:2729–2739
Stroka J, Dasko L, Spangenberg B, Anklam E (2004) Determination of the mycotoxin sterigmatocystin, by thin-layer chromatography and reagent-free derivatisation. J Liq Chromatogr Rel Technol 27:2101–2111
Bleichert M, Eckhardt H-S, Klein K-F, Spangenberg B (2008) A simple and reliable method for quantification of glucosamine in nutritional supplements. J Planar Chromatogr 21:55–59
Caissie GE, Mallet VN (1976) The fluorimetric detection of pesticides on aluminium oxide layers. J Chromatogr 117:129–136
Baeyens WRG, Lin Ling B (1988) Thin layer chromatographic applications with fluorodensitometric detection. J Planar Chromatogr 1:198–213
Prior RL, Wu X, Schaich K (2005) Standardized methods for the determination of antioxidant capacity and phenolics in food and dietary supplements. J Agric Food Chem 53:4290–4302
Hostettmann K, Terreaux Ch, Marston A, Potterat O (1997) The role of planar chromatography in the rapid screening and isolation of bioactive compounds from medicinal plants. J Planar Chromatogr 10:251–257
Yrjönen T, Peiwu Li, Summanen J, Hopia A, Vuorela H (2003) Free radical-scavenging activity of phenolics by reversed-phase TLC. J Am Oil Chem Soc 80:9–14
Cee A (1978) New detection method for compounds with a labile halogen on thin-layer chromatograms. J Chromatogr 150:290–292
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Spangenberg, B., Poole, C.F., Weins, C. (2010). Specific Staining Reactions. In: Quantitative Thin-Layer Chromatography. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-10729-0_7
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