Basic Principles of Planar Chromatography and Its Potential for Hyphenated Techniques

Abstract

Sample preparation, detection, identification, and quantitative determination of biomolecules are presented in this chapter. Advantages of planar chromatography and the basic principles (chambers, sample application, and chromatogram development) are also described. Rapid detection of biomolecules plays a strategical role in their investigation. Hyphenated techniques such as planar chromatography coupled to UV diode array detection and to mass spectrometry provide on-line extensive structural information on the metabolites prior to their isolation. In this chapter, the combination of thin-layer chromatography (TLC) with biomolecules specific detection by diode array scanning (DAD), mass spectrometry (MS), nuclear magnetic resonance (NMR), and Fourier transform infrared spectroscopy (TLC-FTIR) is discussed. In the last part of the chapter, the reader will gain useful information about a recent method of planar chromatography -multidimensional planar chromatography (MD-PC) and information on application of different modes of multidimensional planar chromatography and combination of this technique with diode array detection (MD-PC-DAD) and HPLC-DAD for separation, detection, and qualitative and quantitative determination of biomolecules. Planar chromatography as a pilot technique for HPLC is also described.Biomolecules are widespread throughout the world. It is an organic molecule produced by a living organism, including large polymeric molecules such as proteins, polysaccharides, and nucleic acids as well as small molecules such as a primary metabolites, secondary metabolites, and natural products. Many sample-preparation techniques are used in biomolecule analysis; the method selected depends on the

Современные Методы Анализа Каротиноидов (Обзор)

“…Îäíà-êî íåêîòîðûå ïîëàãàþò, ÷òî îíà ñëóaeèò ïðîñòî äîïîë-íåíèåì ê ÂÝAEÕ, ïîçâîëÿÿ ïðîâîäèòü ëèøü áûñòðûé êà÷åñòâåííûé àíàëèç [23], òàê êàê âîçìîaeíîñòü îêèñ-ëåíèÿ êèñëîðîäîì âîçäóõà íà îòêðûòûõ ïëàñòèíêàõ ïðåïÿòñòâóåò ïîëó÷åíèþ äîñòîâåðíûõ êîëè÷åñòâåí-íûõ äàííûõ [37]. Ïðè ýòîì ñîâåðøåííî íå ó÷èòûâàåò-ñÿ ðàçâèòèå ñîâðåìåííûõ ìåòîäîâ ïðîâåäåíèÿ ÒÑÕ (àâòîìàòèçàöèè íàíåñåíèÿ îáðàçöîâ íà ïëàñòèíêè, ìå-òîäîâ ýëþèðîâàíèÿ, â òîì ÷èñëå ìíîãîñòóïåí÷àòîãî èçîêðàòè÷åñêîãî èëè ãðàäèåíòíîãî, à òàêaeå äâóõìåð-íîãî), ðåãèñòðàöèè è êîëè÷åñòâåííîé îáðàáîòêè õðî-ìàòîãðàìì) [27,28,38], êîòîðûå ïðèâåëè ê ïðåâðàùå-íèþ ÒÑÕ è, îñîáåííî, âûñîêîýôôåêòèâíîé òîíêîñ-ëîéíîé õðîìàòîãðàôèè (ÂÝÒÑÕ) â ýôôåêòèâíûé ìåòîä êà÷åñòâåííîãî è êîëè÷åñòâåííîãî àíàëèçà [32,39]. Ñåãîäíÿ ýòîò ìåòîä õàðàêòåðèçóåòñÿ âîçìîae-íîñòüþ îäíîâðåìåííîãî àíàëèçà çíà÷èòåëüíîãî êîëè-÷åñòâà îáðàçöîâ (íàïðèìåð, äî 72 íà 1 ïëàñòèíêå [28, ñ.…”

“…Ïðèìåíåíèå âìåñòî ñêàíèðóþùèõ äåíñèòîìåòðîâ äèîäíî-ìàòðè÷íîé äåíñèòîìåòðèè õðî-ìàòîãðàìì ðåçêî ñíèaeàåò ïðîäîëaeèòåëüíîñòü àíàëèçà (30 ñ è ìåíåå íà òðåê [46]). Êðîìå òîãî, òàêèå ñêàíåðû â ñî÷åòàíèè ñ îïòîâîëîêîííîé îïòèêîé ïîçâîëÿþò îä-íîâðåìåííî ñ õðîìàòîãðàììîé ðåãèñòðèðîâàòü ñïåê-òðû îáíàðóaeåííûõ ñîåäèíåíèé â äèàïàçîíå 191 -1033 íì [39,40] è ïðîâîäèòü ñðàâíåíèå ýòèõ ñïåêòðîâ ñ áèáëèîòå÷íûìè, îáåñïå÷èâàÿ íàä¸aeíîñòü èäåíòèôèêàöèè è êîëè÷åñòâåííîãî îïðåäåëåíèÿ. Ïðè ýòîì âîçìîaeíà ðåãèñòðàöèÿ òðåõìåðíûõ èçîáðàaeåíèé õðîìàòîãðàìì, îáëåã÷àþùèõ àíàëèç è èíòåðïðåòàöèþ ïîëó÷åííûõ äàííûõ.…”

“…Ïîìèìî äåòåêòîðîâ, èñïîëüçóþùèõ ÓÔ è âèäèìóþ îáëàñòü ñïåêòðà, â ÒÑÕ âñ¸øèðå ïðèìåíÿþòñÿ ÈÊ-Ôóðüå [39,47,48], ßÌÐ [39] è ìàññ-ñïåêòðîñêîïè-÷åñêèå [40] äåòåêòîðû. Ñ èñïîëüçîâàíèåì âñåõ ýòèõ ìåòîäîâ ÒÑÕ ïîçâîëÿåò ðåøàòü ñàìûå ñëîaeíûå çàäà÷è â îáëàñòè àíàëèçà, â òîì ÷èñëå îáðàçöîâ, ñîäåðaeàùèõ êàðîòèíîèäû.…”

Determination of Pesticides in Complex Samples by One Dimensional (1D-), Two-Dimensional (2D-) and Multidimensional Chromatography

“…Planar chromatography is also selected for pesticides screening analysis, because (Tuzimski, 2011b) with multi-wavelength scanning of the chromatograms, spectral data of the analytes can be directly acquired from the TLC plates and can further be compared with the spectra of the analytes from software library; -additional information for structural elucidation can be obtained by planar chromatography combined with MS (fast atom bombardment (FAB) and liquid secondary mass spectrometry (SIMS)); -the whole procedure of chromatographic development can be followed visually, so any distortion of the solvent front, etc., can be observed directly; -the chromatogram can be developed simply by dipping the plate into a mobile phase; -the possibility of two-dimensional development with a single adsorbent; -the possibility of two-dimensional development on e.g., silica -octadecyl silica coupled layers (Multi-K SC5 and CS5 dual phase); -planar chromatography is also the easiest technique which performs multidimensional separation (e.g., by graft chromatography or multidimensional chromatography). Summing up, planar chromatography is one of principal separation techniques, which plays an important role also in environmental analysis.…”

Determination of analytes in medical herbs extracts by SPE coupled with two‐dimensional planar chromatography in combination with diode array scanning densitometry and HPLC‐diode array detector

“…with a narrow zone of SiO 2 and a wide zone of RP-18 (or vice versa) which are commercially available from Whatman (Multi K SC5 or CS5 plates) [17]. Application of MDPC-DAD with NP system on silica and HPLC-DAD with RP system on octadecyl silica was described for correct identification of pesticides in plant extracts on silica plates [17,21]. The application of MDPC combined with different separation systems and modes of chromatogram development is often necessary for performing the separation of more complicated multicomponent mixtures.…”