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We present a two-dimensional (2D) planar chromatographic separation of estrogenic active compounds on RP-18 W (Merck, 1.14296) phase. A mixture of 8 substances was separated using a solvent mix consisting of hexane, ethyl acetate, acetone (55:15:10, v/v) in the first direction and of acetone and water (15:10, v/v) in the second direction. Separation was performed on an RP-18 W plate over a distance of 70 mm. This 2D-separation method can be used to quantify 17α-ethinylestradiol (EE2) in an effect-directed analysis, using the yeast strain Saccharomyces cerevisiae BJ3505. The test strain (according to McDonnell) contains the estrogen receptor. Its activation by estrogen active compounds is measured by inducing the reporter gene lacZ which encodes the enzyme β-galactosidase. This enzyme activity is determined on plate by using the fluorescent substrate MUG (4-methylumbelliferyl-β-d-galactopyranoside).
The use of a TLC scanner can be regarded as a key step in high performance thin layer chromatography (HPTLC). Densitometric measurements transform the substance distribution on a TLC plate into digital computer data. Systems that allow quantitative measurements have been available for many years for either fluorescence or ultraviolet absorption measurements, while lately the reflection analysis mode for both types is the most common application. New scanning approaches are designed to aid the analyst who has common demands for TLC-densitometry without using special data, such as scanned images. Two examples that have been developed lately in the laboratories of the authors are described in this paper. These approaches were developed on the basis of current needs for analysts who employ TLC as a tool in research, as well as in routine analysis. One approach is aimed to support analysts in economically disadvantaged areas, where cost intensive apparatus is unsuitable but trace analysis by simple means is required. The other system, allows the spectral determination of chromatographic spots on TLC plates covering the ultraviolet and visible range, thus, revealing highly desired information for the analyst.
An interlaboratory comparison was carried out to evaluate the effectiveness of a method based on HPTLC in which reagent-free derivatization is followed by UV/fluorescence detection. The method was tested for the determination of sucralose (C12H19C13O8; (2R,3R,4R,5S,6R)-2-[(2R,3S,4S,5S)-2,5-bis(chloromethyl)-3,4-dihydroxyoxolan-2-yl]oxy-5-chloro-6-hydroxymethyl)oxane-3, 4-diol; CAS Registry No. 56038-13-2) in carbonated and still beverages at the proposed European regulatory limits. For still beverages, a portion of the sample was diluted with methanol-water. For carbonated beverages, a portion of the sample was degassed in an ultrasonic bath before dilution. Turbid beverages were filtered after dilution through an HPLC syringe filter. The separation of sucralose was performed by direct application on amino-bonded (NH2) silica gel HPTLC plates (no cleanup needed) with the mobile phase acetonitrile-water. Sucralose was determined after reagent-free derivatization at 190 degrees C; it was quantified by measurements of both UV absorption and fluorescence. The samples, both spiked and containing sucralose, were sent to 14 laboratories in five different countries. Test portions of a sample found to contain no sucralose were spiked at levels of 30.5, 100.7, and 299 mg/L. Recoveries ranged from 104.3 to 124.6% and averaged 112% for determination by UV detection; recoveries ranged from 98.4 to 101.3% and averaged 99.9% for determination by fluorescence detection. On the basis of the results for spiked samples (blind duplicates at three levels), as well as sucralose-containing samples (blind duplicates at three levels and one split level), the values for the RSDr ranged from 10.3 to 31.4% for determinations by UV detection and from 8.9 to 15.9% for determinations by fluorescence detection. The values for the RSDR values ranged from 13.5 to 31.4% for determinations by UV detection and from 8.9 to 20.7% for determinations by fluorescence detection.
A new formula is presented for transforming fluorescence measurements in accordance with Kubelka-Munk theory. The fluorescence signals, the absorption signals, and data from a selected reference are combined in one expression. Only diode-array techniques can measure all the required data simultaneously to linearize fluorescence data correctly. To prove the new theory HPTLC quantification of the analgesic flupirtine was performed over the mass range 300 to 5000 ng per spot. The fluorescence calibration curve was linear over the whole range. The transformation of fluorescence measurements into linear mass-dependent data extends the technique of in-situ fluorescence analysis to the high concentration range. It also extends Kubelka-Munk theory from absorption to fluorescence analysis. The results presented also emphasize the importance of Kubelka-Munk theory for in-situ measurements in scattering media, especially in planar chromatography.
We present a videodensitometric quantification method for methadone in syrup, separated by thin-layer chromatography (TLC). The quantification is based on a derivation reaction with Dragendorf reagent. Measurements were carried out using a 16-bit flatbed scanner. The range of linearity covers two magnitudes of power using the Kubelka-Munk expression for data transformation. The separation method is inexpensive, fast, and reliable.
A Simple and Reliable HPTLC Method for the Quantification of the Intense Sweetener Sucralose®
(2003)
This paper describes a simple and fast thin layer chromatography (TLC) method for the monitoring of the relatively new intense sweetener Sucralose® in various food matrices. The method requires little or no sample preparation to isolate or concentrate the analyte. The Sucralose® extract is separated on amino‐TLC‐plates, and the analyte is derivatized “reagent‐free” by heating the developed plate for 20 min at 190°C. Spots can be measured either in the absorption or fluorescence mode. The method allows the determination of Sucralose® at the levels of interest regarding foreseen European legislation (>50 mg/kg) with excellent repeatability (RSD = 3.4%) and recovery data (95%).
Die quantitative Dünnschichtchromatographie (HPTLC) mit einem Graustufen-Handscanner ist eine preiswerte, schnelle und präzise Methode zur Schwermetallbestimmung. Als Alternative zu teuren Densitometern wird ein Grünlichtscanner mit einer Auflösung von 256 Graustufen benutzt. Die Ortsauflösung beträgt maximal 400 dpi (dots per inch). Die Chromatogramme werden mit 300 dpi aufgenommen. Zur Entwicklung wird eine Camag-Linearkammer verwendet. Zur Probenvorbereitung werden die zu bestimmenden Schwermetallionen bei pH 4,2 mit Dithizon komplexiert. Nur die Metallkationen Zn(2+), Co(2+), Hg(2+), Cd(2+) und Ni(2+) reagieren zu einem farbigen Metallkomplex, wobei sich Zn(2+)- und Co(2+)-Komplexe chromatographisch abtrennen lassen. Nach Komplexierung der Wasserprobe wird mit Essigsäureethylester ausgeschüttelt, Probe- und Standardlösung auf eine Platte aus Kieselgel SI-60 aufgetragen, mit Essigsäureethylester fokussiert und nach der Trocknung der Platte mit Toluol entwickelt. Die HPTLC-Platte wird mit scannereigener Software eingelesen und im PCX-Format (PC PaintBrusch der Fa. ZSoft) auf die Festplatte abgelegt. Zur Auswertung wird eine Leseroutine benutzt. Die ganze Chromatographiebahn ist mit 150 Einzeldioden aufgenommen, die eine Strecke von 48 mm in 564 Einzelmessungen auflösen. Die Summe aller 150 Einzelaufnahmen liefert das Densitogramm aus dem der Schwermetallgehalt bestimmt wird.
We present a video-densitometric quantification method for the pain killer known as diclofenac and ibuprofen. These non-steroidal anti-inflammatory drugs were separated on cyanopropyl bonded plates using CH2Cl2, methanol, cyclohexane (95+5+40, v/v) as mobile phase. The quantification is based on a bio-effective-linked analysis using vibrio fischeri bacteria. Within 10 minutes a CCD-camera registers the white light of the light-emitting bacteria. Diclofenac and ibuprofen effectively suppress the bacterial light emission which can be used for quantification within a linear range of 10 to 2000 ng. The detection limit for ibuprofen is 20 ng and the limit of quantification 26 ng per zone. Measurements were carried out using a 16-bit ST-1603ME CCD camera with 1.56 megapixels [from Santa Barbara Instrument Group, Inc., Santa Barbara, USA]. The range of linearity covers more than two magnitudes because the extended Kubelka-Munk expression is used for data transformation [1]. The separation method is inexpensive, fast and reliable. Ibuprofen is named after its chemical description: iso-butyl-propanoic phenolic acid. Both pain killers are world-widein use and both substances are stable in aqueous solution. Both substances are mainly excreted in the urine.
Diode-array planar chromatography is a versatile tool for identification of pharmaceutical substances In this paper thirty-three compounds with benzodiazepine properties were investigated and the separating conditions for silica gel HPTLC plates and three mobile phases were optimized. Diode-array HPTLC makes it possible to identify all the compounds with high certainty down to a level of 20 ng. An algorithm for spectral recognition which is combined with R F values from the three separation steps into one fit factor is presented. This set of data is unique for each of the compounds investigated and enables unequivocal identification. The method is rapid, inexpensive, and sensitive down to a level of 20 ng mL −1.
High-performance thin-layer chromatography (HPTLC), as the modern form of TLC (thin-layer chromatography), is suitable for detecting pharmaceutically active compounds over a wide polarity range using the gradient multiple development (GMD) technique. Diode-array detection (DAD) in conjunction with HPTLC can simultaneously acquire ultraviolet‒visible (UV‒VIS) and fluorescence spectra directly from the plate. Visualization as a contour plot helps to identify separated zones. An orange peel extract is used as an example to show how GMD‒DAD‒HPTLC in seven different developments with seven different solvents can provide an overview of the entire sample. More than 50 compounds in the extract can be separated on a 6-cm HPTLC plate. Such separations take place in the biologically inert stationary phase of HPTLC, making it a suitable method for effect-directed analysis (EDA). HPTLC‒EDA can even be performed with living organism, as confirmed by the use of Aliivibrio fischeri bacteria to detect bioluminescence as a measure of toxicity. The combining of gradient multiple development planar chromatography with diode-array detection and effect-directed analysis (GMD‒DAD‒HPTLC‒EDA) in conjunction with specific staining methods and time-of-flight mass spectrometry (TOF‒MS) will be the method of choice to find new chemical structures from plant extracts that can serve as the basic structure for new pharmaceutically active compounds.
High performance thin layer chromatography (HPTLC) is a frequently used separation technique which works well for quantification of caffeine and quinine in beverages. Competing separation techniques, e.g. high-performance liquid chromatography (HPLC) or gas chromatography (GC), are not suitable for sugar-containing samples, because these methods need special pretreatment by the analyst. In HPTLC, however, it is possible to separate ‘dirty’ samples without time-consuming pretreatment, because disposable HPTLC plates are used. A convenient method for quantification of caffeine and quinine in beverages, without sample pretreatment, is presented below. The basic theory of in-situ quantification in HPTLC by use of remitted light is introduced and discussed. Several linearization models are discussed.
A home-made diode-array scanner has been used for quantification; this, for the first time, enables simultaneous measurements at different wavelengths. The new scanner also enables fluorescence evaluation without further equipment. Simultaneous recording at different wavelengths improves the accuracy and reliability of HPTLC analysis. These aspects result in substantial improvement of in-situ quantitative densitometric analysis and enable quantification of compounds in beverages.
Die Weltwirtschaftskrise 2008 hat mit ihrer zeitweisen Verknappung von Acetonitril eindringlich gezeigt, dass man nicht nur auf eine einzige chromatographische Methode setzten sollte. Genau dies wird aber im Augenblick getan, denn Industrie und Forschung setzen mehrheitlich auf die High Performance Liquid Chromatography (HPLC) als die Trennmethode ihrer Wahl. Für viele Anwendungen in der Pharmazie, in der Umweltanalytik, der Lebensmittelanalytik, aber auch in der Inprozesskontrolle gibt es mit der Dünnschichtchromatografie eine Alternative.
Die Weltwirtschaftskrise 2008 hat mit ihrer zeitweisen Verknappung von Acetonitril eindringlich gezeigt, dass man nicht nur auf eine einzige chromatographische Methode setzten sollte. Genau dies wird aber im Augenblick getan, denn Industrie und Forschung setzen mehrheitlich auf die High Performance Liquid Chromatography (HPLC) als die Trennmethode ihrer Wahl. Für viele Anwendungen in der Pharmazie, in der Umweltanalytik, der Lebensmittelanalytik, aber auch in der Inprozesskontrolle gibt es mit der Dünnschichtchromatografie eine Alternative.
Fluorescence Enhancement of Pyrene Measured by Thin-Layer Chromatography with Diode-Array Detection
(2003)
In-situ densitometry for qualitative or quantitative purposes is a key step in thin-layer chromatography. It offers a simple way of quantifying by measuring the optical density of the separated spots directly on the plate. A new TLC scanner has been developed which is able to measure TLC plates or HPTLC plates, at different wavelengths simultaneously, without destroying the plate surface. The system enables absorbance and fluorescence measurements in one run. Fluorescence measurements are possible without filters or other adjustments.
The measurement of fluorescence from a TLC plate is a versatile means of making TLC analysis more sensitive. Fluorescence measurements with the new scanner are possible without filters or special lamps. Improvement of the signal-to-noise ratio is achieved by wavelength bundling. During plate scanning the scattered light and the fluorescence are both emitted from the surface of the TLC plate and this emitted light provides the desired spectral information from substances on the TLC plate. The measurement of fluorescence spectra and absorbance spectra directly from a TLC plate is based on differential measurement of light emerging from sample-free and sample-containing zones.
The literature recommends dipping TLC plates in viscous liquids to enhance fluorescence. Measurement of the fluorescence and absorbance spectra of pyrene spots reveals the mechanism of enhancement of plate dipping in viscous liquids—blocked contact of the fluorescent molecules with the stationary phase or other sample molecules is responsible for the enhanced fluorescence at lower concentrations.
In conclusion, dipping in TLC analysis is no miracle. It is based on similar mechanisms observable in liquids. The measured TLC spectra are also very similar to liquid spectra and this makes TLC spec-troscopy an important tool in separation analysis.
A new diode-array scanner in combination with a computer-controlled application system meets all the demands of modern HPTLC measurement. Automatic application, simultaneous measurements at different wavelengths, and different linearization models enable appropriate evaluation of all analytical questions. The theory of error propagation recommends quantification at reflectance values smaller than 0.8; this can be verified only by use of diode-array scanning. The same theory also recommends quantification by use of peak height data, because the theory predicts best precision only for peak height evaluation. Diode-array scanning with reflectance monitoring enables appropriate validation in TLC and HPTLC analysis. All these aspects result in substantial improvement of in-situ quantitative densitometric analysis, and simultaneous recording at different wavelengths opens the way for chemometric evaluation, e.g. peak purity monitoring, which improves the accuracy and reliability of HPTLC analysis.
HPTLC (High Performance Thin Layer Chromatography) is a well known and versatile separation method which shows a lot of advantages and options in comparison to other separation techniques. The method is fast and inexpensive and does not need time-consuming pretreatments. Using fiber-optic elements for controlled light-guiding, the TLC-method was significantly improved: the new HPTLC-system is able to measure simultaneously at different wavelengths without destroying the plate surface or the analytes on the surface. For registration of the sample distribution on a HPTLC-plate we developed a new and sturdy diode-array HPTLC- scanner which allows registration of spectra on the TLC- plates in the range of 198 nm to 610 nm with a spectral resolution better than 1.2 nm. The spatial resolution on plate is better than 160 micrometers . In the spectral mode, the new HPTLC-scanner delivers much more information than the commonly used TLC-scanner. The measurement of 450 spectra of one separation track does not need more than three minutes. However, in the fixed wavelength mode the contour plot can be measured within 15 seconds. In this case, the signal will be summarized and averaged over a spectral range having FWHM from 10 nm to 25 nm depending on the substance under test. The new diode-array HPTLC-scanner makes various chemometric applications possible. The new method can be used easily in clinical diagnostic systems easily, e.g. for blood and uring investigations. In addition, new applications are possible. For example, the rich structured PAHs were studied. Although the separation is incomplete the 16 compounds can be quantified using suitable wavelengths.
In-situ densitometry for qualitative or quantitative purposes is a key step in thin-layer chromatography (TLC). It is a simple means of quantification by measurement of the optical density of the separated spots directly on the plate. A new scanner has been developed which is capable of measuring TLC or HPTLC (high-performance thin-layer chromatography) plates simultaneously at different wavelengths without damaging the plate surface. Fiber optics and special fiber interfaces are used in combination with a diode-array detector. With this new scanner sophisticated plate evaluation is now possible, which enables use of chemometric methods in HPTLC. Different regression models have been introduced which enable appropriate evaluation of all analytical questions. Fluorescent measurements are possible without filters or special lamps and signal-to-noise ratios can be improved by wavelength bundling. Because of the richly structured spectra obtained from PAH, diode-array HPTLC enables quantification of all 16 EPA PAH on one track. Although the separation is incomplete all 16 compounds can be quantified by use of suitable wavelengths. All these aspects are enable substantial improvement of in-situ quantitative densitometric analysis.
In this paper a high-performance thin-layer chromatography (HPTLC) scanner is presented in which a special fibre arrangement is used as HPTLC plate scanning interface. Measurements are taken with a set of 50 fibres at a distance of 400 to 500 μm above the HPTLC plate. Spatial resolutions on the HPTLC plate of better than 160 μm are possible. It takes less than 2 min to scan 450 spectra simultaneously in a range of 198 to 610 nm. The basic improvement of the item is the use of highly transparent glass fibres which provide excellent transmission at 200 nm and the use of a special fibre arrangement for plate illumination and detection.
Nativ-organische Abfälle bilden mit ca. 30 Gew. % den Hauptteil des Hausmülls. Daher leistet die Bioabfallkompostierung einen bedeutenden Schritt hin zu einer sinnvollen Abfallverwertung. Bundesweit werden derzeit jährlich etwa 750 000 t Bioabfallkompost erzeugt. Mit Ausnahme von sieben Land- und Stadtkreisen planen die Landkreise Baden-Württembergs die getrennte Sammlung von Bioabfällen: Einwohner von 23 Landkreisen waren 1993, zum Teil versuchsweise, an Biotonnen angeschlossen. Die Notwendigkeit der Bioabfallkompostierung scheint außer Frage zu stehen, intensiv erörtert werden jedoch die Verfahrenskonzepte, die der Bioabfallkompostierung zugrunde liegen. Kern der Diskussion ist, ob einfache Kompostierungsverfahren wie die dezentrale Kompostierung den technisch aufwendigeren Verfahren zentraler Anlagen gleichwertig sind.
Als erster Landkreis in Baden Württemberg hat der Landkreis Sigmaringen bei der Entsorgung des bei 120 000 Einwohnern im Kreis anfallenden organischen Abfalls neue Wege beschritten und die Landwirtschaft in den Stoffkreislauf miteinbezogen, anstatt das anfallende Material zu deponieren. Auf dem Hintergrund der TA-Siedlungsabfall, die vorschreibt, daß nach einer Übergangszeit organische Abfälle nicht mehr deponiert werden dürfen, sondern kompostiert oder thermisch behandelt werden müssen, hat der Landkreis Sigmaringen im Jahr 1992 ein Konzept zur dezentralen Kompostierung verabschiedet.
We present a video-densitometric high-performance thin-layer chromatography (HPTLC) quantification method for patulin in apple juice, developed in a vertical chamber from the starting point to a distance of 50 mm, using MTBE, n-pentane (9 + 5, v/v) as mobile phase. After separation the plate is sprayed with methyl-benzothiazolinone hydrazone hydrochloride monohydrate (MBTH) solution (40 mg in 20 mL methanol) and heated at 105 °C for 15 min. Patulin zones are transformed into yellow spots. The quantification is based on direct measurements using an inexpensive 48-bit flatbed scanner for color measurements (in red, green, and blue). Evaluation of the blue channel makes the measurements very specific. Quantification in fluorescence was also done by use of a 16-bit CCD-camera and UV-366 nm illumination as well as using a HPTLC DAD-scanner. For linearization the extended Kubelka–Munk expression for data transformation was used. The range of linearity covers more than two magnitudes and lies between 5 and 800 ng patulin. The extraction of 20 g apple juice and an extract application on plate up to 50 µL allows statistically defined checking the limit of detection (LOD) of 50 ng patulin per track, which is equivalent to 50 µg patulin per kg apple juice.
Poröse Massen oder Formkörper aus anorganischen polymeren und deren Herstellung (EP000002958875A1)
(2015)
Poröse Massen oder Formkörper aus anorganischen polymeren und deren Herstellung (WO002014127762A1)
(2014)
Es wird ein Verfahren zur Herstellung einer porösen Masse oder eines porösen Formkörpers aus anorganischem Polymer beschrieben, bei dem Wasserglas mit einem Carbonat in bestimmten Mengen gehärtet wird und dabei verschiedene weitere Stoffe zugesetzt werden können. Außerdem werden poröse Massen und Formkörper, die mit diesem Verfahren erhältlich sind, beschrieben und deren Verwendung.
The NaSiO Institute (Institute for Sustainable Silicate Research in Offenburg, https://inasio.hs-offenburg.de/) has been working for years on climate-friendly alternatives to insulation materials and inorganic binders, as well as the reasonable use of construction waste in the building industry. The aim of research is to realize the enormous CO 2 saving potential of the construction sector worldwide. A stopping of climate heating will only succeed if these climate-friendly alternatives are used in the construction industry. This is the only way to realize the enormous CO2 savings that will be needed in future to comply with the Paris Agreement.
An Extraction Method for 17α-Ethinylestradiol from Water using a new kind of monolithic Stir-bar
(2015)
Es wird ein Verfahren zur Herstellung eines nicht geschäumten porösen monolithischen oder faserförmigen Produkts aus anorganischem Polymer beschrieben, bei dem Wasserglas mit einem Carbonat und/oder Amid in bestimmten Mengen in Gegenwart von Wasser gehärtet wird. Außerdem werden monolithische Chromatographiesäulen und -platten, geträgerte Metall-Katalysatoren und Vliese beschrieben.
Described is a solid body formed with Si, Al, Ca, O and at least one of Na and K, said body exhibiting in the 27Al MAS NMR spectrum a signal additional to the 27Al MAS NMR spectrum of pure calcium aluminate, with a chemical shift sited between that of the main peak of calcium aluminate and the peak next upfield to the main peak of calcium aluminate. Possible uses of the solid body include use as a building material with aggregates, as a coating, as an adhesive for joining two components for sanitary ceramic units, for high-temperature applications, for renovating existing edifices, especially for underwater renovation, for the erection and/or repair of built structures, particularly when high compressive strengths are needed or chemically aggressive conditions arise. It can be produced by bringing waterglass, sodium hydroxide and/or potassium hydroxide, calcium aluminate, one or more aggregates and optionally water, especially sea water, into contact, even at temperatures below 0°C without heating.
A simple Method for quantifying Triazine Herbicides using Thin-Layer Chromatography and a CCD-Camera
(2010)
We present a video-densitometric quantification method for the triazine herbicides atraton, terbumeton, simazine, atrazine, and terbutylazine. Triazine herbicides were separated on silica gel using methyl-t-butyl ether, cyclohexane (1 + 1, v/v) as mobile phase. The quantification is based on a derivation reaction using chlorine and starch-iodine which forms red-brown triazine zones. Measurements were carried out using a 16 bit ST-1603ME CCD camera with 1.56 megapixel from Santa Barbara Instrument Group, Inc., Santa Barbara, USA. A white LED was used for illumination purposes. The range of linearity covers two magnitudes using the (1/R-1) expression data transformation. The signal-to-noise ratio increases directly linearly with the measurement time. The separation method is cheap, fast and reliable.
Melamine (1,3,5-triazine-2,4,6-triamine or cyanuramide, C3H6N6) is a trimer of cyanamide, with a 1,3,5-triazine skeleton (Figure 3.5-1). The molecule contains 66% nitrogen by mass and, if mixed with resins, has fire retardant properties due to its release of nitrogen gas when burned or charred. The word melamine (from German) is a combination of the word melam (which is a distillation derivative of ammonium thiocyanate) and amine [1]. Melamine is also a metabolite of cyromazine, an insecticide in which the proton of an NH2-group is substituted by a cyclopropyl group.
We present a densitometric quantification method for triclosan in toothpaste, separated by high-performance thin-layer chromatography (HPTLC) and using a 48-bit flatbed scanner as the detection system. The sample was band-wise applied to HPTLC plates (10 × 20 cm), with fluorescent dye, Merck, Germany (1.05554). The plates were developed in a vertical developing chamber with 20 min of chamber saturation over 70 mm, using n-heptane–methyl tert-butyl ether–acetic acid (92:8:0.1, V/V) as solvent. The RF value of triclosan is hRF = 22.4, and quantification is based on direct measurements using an inexpensive 48-bit flatbed scanner for color measurements (in red, green, and blue) after plate staining with 2,6-dichloroquinone-4-chloroimide (Gibbs' reagent). Evaluation of the red channel makes the measurements of triclosan very specific. For linearization, an extended Kubelka–Munk expression was used for data transformation. The range of linearity covers more than two orders of magnitude and is between 91 and 1000 ng. The separation method is inexpensive, fast and reliable.
HPTLC (High Performance Thin Layer Chromatography) is a well known and versatile separation method which shows many advantages when compared to other separation techniques. The method is fast and inexpensive and does not need time-consuming pretreatments. For visualisation of the sample distribution on a HPTLC-plate we developed a new and sturdy HPTLC-scanner. The scanner allows simultaneous registrations of spectra in a range from 198 nm to 612 nm with a spectral resolution of better than 0.8 nm. The on-plate spatial resolution is better than 160 μm. The measurement of 450 spectra in one separation track does not need more than two minutes. The new diode-array scanner offers a fast survey over a TLC-separation and makes various chemometric applications possible. For compound identification a cross-correlation function is described to compare UV sample spectra with appropriate library data. The cross-correlation function herein described can also be used for purity testing. Unresolved peaks can be virtually separated by use of a least squares fit algorithm. In summary, the diode arry system delivers much more information than the commonly used TLC-scanner.
Vorgestellt wird die Dioden-Array-Dünnschichtchromatographie als eine moderne und preiswerte Messmethode zur densitometrischen Erfassung von Substanzen auf einer DC- oder einer HPTLC-Platte. Sicher identifizierbar sind auch Substanzen mit schwachem Chromophor. Die Kubelka-Munk-Gleichung beschreibt einen linearen Zusammenhang zwischen Remissionslicht und lichtabsorbierender Stoffmenge auf der Platte. Die Auswertung im Spektralbereich von 316 bis 334 nm zeigt den Zusammenhang zwischen transformiertem Messsignal und aufgetragener Substanzmasse. Die schnelle Aufnahme von UV/vis-Spektren eröffnet der HPTLC den gesamten Bereich der Methodenvalidierung auf dem Niveau, auf welchem heute die HPLC-Analytik durchgeführt wird.
Rudolf E. Kaiser
(2020)
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Quantitative Bestimmung von Clozapin im Serum mittels Dioden-Array Dünnschichtchromatographie
(2003)
In thin-layer chromatography the development step distributes the sample throughout the layer, a process which strongly affects the reflection signals. The essential requirement for quantitative thinlayer chromatography is not a constant sample concentration but constant sample distribution in each sample spot. This makes evaporation of the mobile phase extremely important, because all tracks of a TLC plate must be dried uniformly. This paper shows that quantitative TLC is possible even if the concentration of the sample is not constant throughout the layer or if the distribution of the sample is not known. With uniform sample distribution, classical Kubelka-Munk theory is valid for isotropic scattering only. In the absence of this constraint classical Kubelka-Munk theory must be extended to situations where scattering is asymmetric. This can be achieved by modification of the original Kubelka-Munk equation. Extended theory is presented which is not only capable of describing asymmetrical scattering in TLC layers but also includes a formula for absorption and fluorescence in diode-array TLC. With this new theory all different formulas for diode-array thin-layer chromatographic evaluation are combined in one expression.
Thin-layer chromatography (TLC) is a well-established and widely used separation technique. Most undergraduate students of chemistry or food science used TLC as a primitive separation tool, which does not need more than small pieces of TLC plates, a glass jar and some solvents. TLC has evolved from a simple separation method of the past into an instrumental technique that offers automation, reproducibility and accurate quantification for a wide variety of applications [1]. The use of modern 10*10 cm TLC plates with narrow particle size distribution is called high performance thin layer chromatography (HPTLC), to distinguish the method from the use of traditional 20 20 cm TLC plates.
A 2D-separation of 16 polyaromatic hydrocarbons (PAHs) according to the Environmental Protecting Agency (EPA) standard was introduced. Separation took place on a TLC RP-18 plate (Merck, 1.05559). In the first direction, the plate was developed twice using n-pentane at −20°C as the mobile phase. The mixture acetonitrile-methanol-acetone-water (12:8:3:3, v/v) was used for developing the plate in the second direction. Both developments were carried out over a distance of 43 mm. Further on in this publication, a specific and very sensitive indication method for benzo[a]pyrene and perylene was presented. The method can detect these hazardous compounds even in complicated PAH mixtures. These compounds can be quantified by a simple chemiluminescent reaction with a limit of detection (LOD) of 48 pg per band for perylene and 95 pg per band for benzo[a]pyrene. Although these compounds were separated from all other PAHs in the standard, a separation of both compounds was not possible from one another. The method is suitable for tracing benzo[a]pyrene and/or perylene. The proposed chemiluminescence screening test on PAHs is extremely sensitive but may indicate a false positive result for benzo[a]pyrene.
The main focus of this chapter is the theoretical and instrumental processes that underpin densitometric methods widely used in thin-layer chromatography (TLC). Densitometric methods include UV–vis, luminescence, and fluorescence optical measurements as well as infrared and Raman spectroscopic measurements. The chapter is divided in two general parts: a theoretical part and a practical part. The systems for direct radioactivity measurements and the combination of TLC with mass spectrometry are also discussed. All these systems allow measuring an intensity distribution directly on a TLC plate. We call this “in situ detection” because no analyte is removed from the plate.
The main focus of this chapter is the theoretical and instrumental processes that underpin densitometric methods widely used in thin-layer chromatography (TLC). Densitometric methods include UV–vis, luminescence and fluorescence optical measurements as well as infrared and Raman spectroscopic measurements. The chapter is divided in two general parts: a theoretical part and a practical part. The systems for direct radioactivity measurements and the combination of TLC with mass spectrometry are also discussed. All these systems allow measuring an intensity distribution directly on a TLC plate. We call this “in situ detection” because no analyte is removed from the plate.
Wir präsentieren die weltweit erste Auswertung einer zweidimensional entwickelten HPTLC-Platte (2D-HPTLC) mit Hilfe eines Diodenarray Scanners. Das System erreicht eine räumliche Plattenauflösung von 250 µm. Es können Absorptions- und Fluoreszenzspektren im Wellenlängenbereich von 190 bis 1000 nm mit einer spektralen Auflösung von besser als 1 nm gemessen werden. Eine Trennzahl von 259 wurde erreicht. Damit zeigt die Methode bessere Trenneigenschaften als die meisten HPLC-Systeme. Der Nachteil der 2D-Auswertung ist der hohe Zeitbedarf von über 3 Stunden für eine Plattenmessung.
Das einzige deutschsprachige Buch zur quantitativen Dünnschichtchromatographie auf dem Markt
Für Wissenschaftler und Anwender: von der vollständigen Behandlung der Theorie bis hin zu Entwicklungs- und Auswertetechniken
Auch für Einsteiger geeignet, da das Buch entsprechend der Arbeitsschritte einer DC-Analyse aufgebaut ist
Alle Materie strebt nach maximaler Unordnung. Diese Erkenntnis wird durch die thermodynamische Funktion der Entropie beschrieben. Auch bei jeglicher Art menschlichen Handelns wird die Entropie immer erhöht. Wird in der Technik Materie in geordnete Formen gebracht (z. B. beim Herstellen von Pfandflaschen), findet in diesem Produkt eine Entropieerniedrigung statt. Gleichzeitig wird aber an anderer Stelle die Unordnung beträchtlich vergrößert. Diese Entropieerhöhung nennen wir Abfall. Jede Entropieerhöhung ist mit dem Verbrauch wertvoller Ressourcen verbunden. Durch eine optimale Recyclingtechnik kann einer Entropieerhöhung von Materie entgegengearbeitet werden. Aber nur Recyclingraten von über 90 % erlauben eine wirksame Streckung der Ressourcen.
Two solvent mixtures for high-performance thin-layer chromatographic (HPTLC) separation of some compounds showing estrogenic activity in the yeast estrogen screen (YES) assay are presented. The new method, planar yeast estrogen screen (pYES) combines the YES assay and a chromatographic separation on silica gel HPTLC plates with the performance of the YES assay. For separation, the analytes were applied bandwise to HPTLC plates (10 × 20 cm) with fluorescent dye (Merck, Germany). The plates were developed in a vertical developing chamber after 30 min of chamber saturation over a separation distance of 70 mm, using cyclohexane‒methyl-ethyl ketone (2:1, V/V) or cyclohexane‒CPME (3:2, V/V) as solvents. Both solvents allow separation of estriol, daidzein, genistein, 17β-estradiol, 17α-ethinyl estradiol, estrone, 4-nonylphenol and bis(2-ethylhexyl) phthalate.
An algorithm is presented that has successfully been utilized in practice for several years. It improves data analysis in chromatography. The program runs in an extremely reliable way and evaluates chromatographic raw data with an acceptable error. The algorithm requires a minimum of preliminaries and integrates even unsmoothed noisy data correctly.
Improved separation of highly toxic contact herbicides paraquat (1,1′-dimethyl-4-4′-bipyridinium), diquat (6,7-dihydrodipyridol[ 1,2-a:2′,1′-c]pyrazine-5,8-di-ium), difenzoquat (1,2-dimethyl-3,5-diphenyl-1H-pyrazolium-methyl sulfate), mepiquat (1,1-dimethyl-piperidinium), and chloromequat (2-chloroethyltrimethylammonium) were presented by high-performance thin-layer chromatography (HPTLC). The quantification is based on a derivatization reaction, using sodium tetraphenylborate. Measurements were made in the wavelength range from 500 to 535 nm, using a light-emitting diode (LED) for excitation purposes, which emits very dense light at 365 nm. For calculations, a new theory of standard addition method was used, thus leading to a minimal error if exactly the same amount of sample content is added as a standard. The method provides a fast and inexpensive approach to quantification of the five most important quats used for plant protection purposes. The method works reliably because it takes into account losses during pre-treatment procedure. The method meets the European legislation limits for paraquat and diquat in drinking water according to United States Environmental Protection Agency (US EPA) method 549.2 which are 680 ng L−1 for paraquat and 720 ng L−1 for diquat. The method of standard addition in planar chromatography can be beneficially used to reduce systematic errors. Although recovery rates of 33.7% to 65.2% are observed, calculated contents according to the method of standard addition lie between 69% and 127% of the theoretical amounts.
Editorial
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We present a video-densitometric quantification method for the pain killer known as diclofenac and ibuprofen. These non-steroidal anti-inflammatory drugs were separated on cyanopropyl bonded plates using CH2Cl2, methanol, cyclohexane (95 + 5 + 40, v/v) as mobile phase. The quantification is based on a bio-effective-linked analysis using Vibrio fisheri bacteria. Within 10 min a CCD-camera registered the white light of the light-emitting bacteria. Diclofenac and ibuprofen effectively suppressed the bacterial light emission which can be used for quantification within a linear range of 10 to 2000 ng. The detection limit for ibuprofen is 20 ng and the limit of quantification 26 ng per zone. Measurements were carried out using a 16-bit ST-1603ME CCD camera with 1.56 megapixels (from Santa Barbara Instrument Group, Inc., Santa Barbara, USA). The range of linearity covers more than two magnitudes because the extended Kubelka-Munk expression is used for data transformation. The separation method is inexpensive, fast, and reliable.
We present a video-densitometric quantification method in combination with diode-array quantification for the methyl-, ethyl-, propyl-, and butylparaben in cosmetics. These parabens were separated on cyanopropyl bonded plates using water-acetonitrile-dioxane-ethanol-NH3 (25%) (8:2:1:1:0.05, v/v) as mobile phase. The quantification is based on UV-measurements at 255 nm and a bioeffectively-linked analysis using Vibrio fischeri bacteria. Within 5 min, a Tidas S 700 diode-array scanner (J&M, Aalen, Germany) scans 8 tracks and thus measures in total 5600 spectra in the wavelengths range from 190 to 1000 nm. The quantification range for all these parabens is from 20 to 400 ng per band, measured at 255 nm. In the V. fischeri assay a CCD-camera registers the white light of the light-emitting bacteria within 10 min. All parabens effectively suppress the bacterial light emission which can be used for quantifying within a linear range from 100 to 400 ng. Measurements were carried out using a 16-bit MicroChemi chemiluminescence system (biostep GmbH, Jahnsdorf, Germany), using a CCD camera with 4.19 megapixels. The range of linearity is achieved because the extended Kubelka-Munk expression was used for data transformation. The separation method is inexpensive, fast, and reliable.
We present a two-dimensional (2D) planar chromatographic separation method for phytoestrogenic active compounds on RP-18 W (Merck, 1.14296) phase. It could be shown that an ethanolic extract of liquorice (Glycyrrhiza glabra) roots contains four phytoestrogenic active compounds. As solvent, in the first direction, the mix of hexane, ethyl acetate, and acetone (45:15:10, v/v) was used, and, in the second direction, that of acetone and water (15:10, v/v) was used. After separation, a modified yeast estrogen screen (YES) test was applied, using the yeast strain Saccharomyces cerevisiae BJ3505. The test strain (according to McDonnell) contains the estrogen receptor. Its activation by estrogen active compounds is measured by inducing the reporter gene lacZ which encodes the enzyme β-galactosidase. This enzyme activity is determined on plate by using the fluorescent substrate MUG (4-methylumbelliferyl-β-d-galactopyranoside). The enzyme can also hydrolyse X-β-Gal (5-bromo-4-chloro-3-indoxyl-β-d-galactopyranosid) into β-galactose and 5-bromo-4-chloro-3-indoxyl. The indoxyl compound is oxidized by oxygen forming the deep-blue dye 5,5β-dibromo-4,4β-dichloro-indigo which allows to detect phytoestrogenic activity more specific in the presence of native fluorescing compounds.
We present a planar chromatographic separation method for the phytoestrogenic active compound equol, separated on RP-18 W (Merck, 1.14296) phase. It could be shown that an ethanolic cattle manure extract contains this phytoestrogenic active compound to a larger amount. As solvents for the mobile phase, hexane, ethyl acetate, and acetone (45:15:10, v/v); acetone and water (15:10, v/v); and n-hexane, CH2Cl2, ethyl acetate, methanol, and formic acid (40:40:20:5:1, v/v) have been used. After separation, a modified yeast estrogen screen (YES) test was applied, using the yeast strain Saccharomyces cerevisiae BJ3505 containing an estrogen receptor. Its activation by equol induces the reporter gene lacZ which encodes the enzyme β-galactosidase. The enzyme activity is measured directly on the TLC plate by using the substrate MUG (4-methylumbelliferyl-β-d-galactopyranoside) or the substrate X-β-Gal (5-bromo-4-chloro-3-indoxyl-β-d-galactopyranoside). β-Galactosidase cleaves MUG into a fluorescing compound. X-β- Gal is also hydrolyzed and then oxidized by oxygen forming the deep-blue dye 5,5′-dibromo-4,4′-dichloro-indigo. Both reactions in combination with a thin-layer chromatography (TLC) separation allow very specific detecting of equol in cattle manure, although that is a very challenging matrix. Preliminary results show that the average content of equol in liquid manure is roughly 60 μg g−1. The value for urine is 50 μg mL−1.
We present a two dimensional (2D) planar chromatographic separation of estrogenic active compounds on RP-18 (Merck, 1.05559) and silica gel (Merck, 1.05721) phase. A mixture of 13 substances was separated using a solvent mix consisting of methanol–acetonitrile–water (2 + 2 + 1, v/v/v) on RP-18 phase in the first direction and cyclohexane–butylacetate–methanol (8 + 6 + 1, v/v/v) in the second direction on silica gel plate. Both developments were carried out over a distance of 70 mm. We used the grafted method to combine both plates in a 2D-separation. This 2D-separation method can be used to quantify 17α-ethinylestradiol (EE2) in an effect-directed analysis using the yeast strain Saccharomyces cerevisiae BJ3505. The test strain (according to McDonnell) contains the estrogen receptor. Its activation by estrogen active compounds is measured by inducting the reporter gene lacZ that encodes the enzyme ß-galactosidase. This enzyme activity is determined on plate by using the fluorescent substrate MUG (4-methylumbelliferyl ß-D-galactopyranoside).
A Validated Quantification of Sudan Red Dyes in Spicery using TLC and a 16-bit Flatbed Scanner
(2018)
We present a video-densitometric quantification method for Sudan red dyes in spices and spice mixtures, separated by TLC. Application was done band-wise in small dots using a 5 μL glass pipette. For separation, the RP-18 plates (20 × 20 cm with fluorescent dye; Merck, Germany, 1.05559) were developed in a vertical developing chamber without vapor saturation from the starting point to a distance of 70 mm by using acetonitrile, methanol, and aqueous ammonia solution (25%; 8 + 1.8 + 0.2, v/v) as mobile phase. The quantification is based on direct measurements using an inexpensive 16-bit flatbed scanner for color measurements (in red, green, and blue). Evaluation of only the green channel makes the measurements very specific. For linearization, an extended Kubelka-Munk expression for data transformation was used. The range of linearity covers more than two magnitudes and lies between 20 and 500 ng. The extraction from a 2 g sample with acetonitrile, evaporation, and reconstitution to 200 μL with methanol and the band-wise application (7 mm) of a 10 μL sample allows a statistically defined LOD of less than 500 ppb of Sudan red dyes. To perform the analysis, a separation chamber, RP-18 plates, 5 μL glass pipettes, and a 16-bit flatbed scanner for 105 € are needed; therefore, the separation method is inexpensive, fast, and reliable.
Quantification of astaxanthin in salmons by chemiluminescence and absorption after TLC separation
(2018)
Astaxanthin is a keto-carotenoid, belongs to the chemical class of terpenes and is a yellow lipid soluble compound. The compound is present in marine animals like salmons and crustacean. Its colour is due to conjugated double bonds and these double bonds are responsible for its antioxidant effect. Its antioxidant activity is ten times stronger than other carotenoids and nearly 500 fold stronger than vitamin-E. We present a new thin layer chromatography (TLC) method to measure astaxanthin on TLC-plates (Merck, 1.05554) in the visible absorption range as well as by using chemiluminescence. For separation a solvent mixture of cyclohexane and acetone (10 + 2.4, v/v) was used. The RF-value of astaxanthin is 0.14.The limit of detection in vis-absorption is 64 ng / band and the limit of quantification is 92 ng/band. In chemiluminescence the values are 90 ng / band and 115 ng/band. The method offers two independently working measurement modes on a single plate which increase the accuracy of the quantification.
We will present the first example of a two-dimensional scanned TLC-plate, measured by use of a diode-array scanner. A spatial resolution of 250 µm was achieved on plate. The system provides real 2D fluorescence and absorption spectra in the wavelength-range from 190 to 1000 nm with a spectral resolution of greater than 1 nm. A mixture of 12 sulphonamides was separated by using a cyanopropyl-coated silica gel plate (Merck, 1.16464) with the solvent mix of methyl tert-butyl ether-methanol-dichloromethane-cyclohexane-NH3 (25%) (48:2:2:1:1, v/v) in the first and with a mixture of water-acetonitrile-dioxane-ethanol (8:2:1:1, v/v) in the second direction. Both developments were carried out over a distance of 70 mm. A separation number (spot capacity) of 259 was calculated. We discussed a new formula for its calculation in 2D-TLC separations. The drawback of this method is that measuring a 2D-TLC plate needs more than 3 h measurement time.
Limits of quantification of some neonicotinoid insecticides measured by thin-layer chromatography
(2012)
A simple method to quantify the neonicotinoid insecticides nitenpyram, thiamethoxam, acetamiprid, imidacloprid, thiacloprid and clothianidin directly on an HPTLC-plate is presented. As stationary phase silica gel 60 RP-18WF254 s plates were used and a mixture of methyl-t-butyl ether, 2-butanone, NH3 (25%) (5 + 2+0.1, v/v) was used as solvent. All neonicotinoid insecticides show light absorptions below 300 nm. The calculated limits of quantification (LOQ) by UV-detection are in the range from 12 ng to 26 ng on plate depending on the different insecticides.Nitenpyram can be stained using fast blue salt B, forming red zones. The observed LOQ is 25 ng on plate. Acetamiprid can be specifically stained using phenylglyoxylic acid forming a yellow/green fluorescent compound. The LOQ is 52 ng per spot.The compounds thiamethoxam, acetamiprid, thiacloprid and clothianidin can be transformed into blue fluorescing zones, using a relatively new staining solution. This consists of tetraphenylborate and HCl. This is the first publication mentioning that neonicotinoids undergo this reaction. The calculated limits of quantification are in the range from 10 ng to 27 ng on plate.A simple pre-treatment procedure using an acetonitrile extraction and a Chromabond SiOH clean up procedure leads to overall LOQs for bee samples of 48 to 108 µg/Kg. The method can be used to measure neonicotinoid contaminations of bees.
Eine einfache Bestimmung von Mineraloel-KWstoffen - Ersatz des FCKW-haltigen Extraktionsmittels
(1996)
Die Messung von KWstoffen in Abwaessern nach DIN ist eine in der Umweltanalytik haeufig geforderte Bestimmung. Die Abwasserprobe wird dabei mit 1,1,2-Trichlortrifluorethan extrahiert. Anschliessend wird der Extrakt mittels IR-Spektroskopie vermessen. Neben einigen Schwaechen ist bei dieser Bestimmungsmethode besonders die Verwendung des ozonschaedigenden FCKW-Loesemittels heute nicht mehr zeitgemaess. - Die Verf. beschreiben ein schnelles robustes Bestimmungsverfahren, das alle Schwaechen der alten Methode vermeidet.
We present an improved quantification method for urethane found in spirits. The quantification is based on a derivatization reaction using cinnamaldehyde in combination with phosphoric acid. Measurements were carried out in the wavelength range from 445 to 460 nm using a diode-TLC device. An LED was used for illumination purposes. It emits very dense light at 365 nm. The quantification range of urethane is in the lower ng range. By applying 20 µL of sprits, the urethane quantification range is from 320 µg/L to 8.1 mg urethane per litre of spirit. The range of linearity covers nearly two magnitudes. The method is cheap, fast and reliable, and is able to monitor all European legislation limits without time-consuming sample pre-treatments.
BioPower
(2009)
Das Projekt BioPower ist eine Kooperation des Instituts für Angewandte Forschung (IAF) der Hochschule Offenburg mit dem Institut für Mikrosystemtechnik (IMTEK) der Universität Freiburg. Es handelt sich um den Versuch, die im Körper vorhandenen Energiequellen sozusagen direkt anzuzapfen, um sie für technische Zwecke zu nutzen. Von den vielen bestehenden Möglichkeiten konzentriert sich die Forschung hier auf die Nutzung der Glukose im Blut, die auch sonst als Energieträger zur Versorgung der Zellen im Körper dient.
Vorrichtung (2) zur Analyse von Urin, umfassend: – eine Zuführ- und Abführeinrichtung (7), welche zur Zuführung einer bestimmten Urinmenge in eine wenigstens einen Analysebereich (8) aufweisende Analysekammer (9) eines Urinteststreifens (10) und zur Abführung einer bestimmten Urinmenge aus einer wenigstens einen Analysebereich (8) aufweisenden Analysekammer (9) eines Urinteststreifens (10) eingerichtet ist, wobei die Zuführ- und Abführeinrichtung (7) wenigstens ein bewegbar gelagertes Zuführ- und/oder Abführelement (28, 29) zum Zuführen einer bestimmten Urinmenge in einen Zuführbereich (33) der Analysekammer (9) des Urinteststreifens (10) und/oder zum Abführen einer bestimmten Urinmenge aus einem Abführbereich (34) der Analysekammer (9) des Urinteststreifens (10) aufweist, und – eine Erfassungseinrichtung (11), welche zur Erfassung einer zumindest abschnittsweisen Änderung wenigstens eines optisch erfassbaren Parameters, welcher sich in Abhängigkeit der Zusammensetzung einer diesen kontaktierenden Urinmenge optisch erfassbar verändert, des oder eines entsprechenden Analysebereichs (8) des oder eines entsprechenden Urinteststreifens (10) sowie zur Erzeugung einer Erfassungsinformation, welche wenigstens einen optisch erfassten Parameter des oder eines entsprechenden Analysebereichs (8) oder eine Änderung eines solchen beschreibt, eingerichtet ist.
Vorrichtung (2) zur Analyse von Urin, umfassend: – eine Zuführ- und Abführeinrichtung (7), welche zur Zuführung einer bestimmten Urinmenge in eine wenigstens einen Analysebereich (8) aufweisende Analysekammer (9) eines Urinteststreifens (10) und zur Abführung einer bestimmten Urinmenge aus einer wenigstens einen Analysebereich (8) aufweisenden Analysekammer (9) eines Urinteststreifens (10) eingerichtet ist, wobei die Zuführ- und Abführeinrichtung (7) wenigstens ein bewegbar gelagertes Zuführ- und/oder Abführelement (28, 29) zum Zuführen einer bestimmten Urinmenge in einen Zuführbereich (33) der Analysekammer (9) des Urinteststreifens (10) und/oder zum Abführen einer bestimmten Urinmenge aus einem Abführbereich (34) der Analysekammer (9) des Urinteststreifens (10) aufweist, und – eine Erfassungseinrichtung (11), welche zur Erfassung einer zumindest abschnittsweisen Änderung wenigstens eines optisch erfassbaren Parameters, welcher sich in Abhängigkeit der Zusammensetzung einer diesen kontaktierenden Urinmenge optisch erfassbar verändert, des oder eines entsprechenden Analysebereichs (8) des oder eines entsprechenden Urinteststreifens (10) sowie zur Erzeugung einer Erfassungsinformation, welche wenigstens einen optisch erfassten Parameter des oder eines entsprechenden Analysebereichs (8) oder eine Änderung eines solchen beschreibt, eingerichtet ist.
Vorrichtung (2) zur Analyse von Urin, umfassend: – eine Zuführ- und Abführeinrichtung (7), welche zur Zuführung einer bestimmten Urinmenge in eine wenigstens einen Analysebereich (8) aufweisende Analysekammer (9) eines Urinteststreifens (10) und zur Abführung einer bestimmten Urinmenge aus einer wenigstens einen Analysebereich (8) aufweisenden Analysekammer (9) eines Urinteststreifens (10) eingerichtet ist, wobei die Zuführ- und Abführeinrichtung (7) wenigstens ein bewegbar gelagertes Zuführ- und/oder Abführelement (28, 29) zum Zuführen einer bestimmten Urinmenge in einen Zuführbereich (33) der Analysekammer (9) des Urinteststreifens (10) und/oder zum Abführen einer bestimmten Urinmenge aus einem Abführbereich (34) der Analysekammer (9) des Urinteststreifens (10) aufweist, und – eine Erfassungseinrichtung (11), welche zur Erfassung einer zumindest abschnittsweisen Änderung wenigstens eines optisch erfassbaren Parameters, welcher sich in Abhängigkeit der Zusammensetzung einer diesen kontaktierenden Urinmenge optisch erfassbar verändert, des oder eines entsprechenden Analysebereichs (8) des oder eines entsprechenden Urinteststreifens (10) sowie zur Erzeugung einer Erfassungsinformation, welche wenigstens einen optisch erfassten Parameter des oder eines entsprechenden Analysebereichs (8) oder eine Änderung eines solchen beschreibt, eingerichtet ist.
Phenolic compounds, such as flavonoids and phenolic acids, are very important substances that occur in various medicinal plants. They show different pharmacological activities which might be useful in the therapy of many diseases. Phenolic compounds have achieved an increasing interest over the last years because these compounds are easily oxidized and, thus, act as strong antioxidants. We present the chemiluminescence of different phenolic compounds measured directly on high-performance thin-layer chromatography LiChrospher® plates using the oxalic acid derivative bis(2,4,6-trichlorophenyl) oxalate (TCPO) in conjunction with H2O2. Our results indicate that chemiluminescence intensity increases with an ascending number of phenolic groups in the molecule. The method can be used to detect phenolic compounds in beverages like coffee, tea, and wine.
A diode array HPTLC method for dequalinium chloride in pharmaceutical preparations is presented. For separation a Nano TLC silica gel plate (Merck) is used with the mobile phase methanol-7.8% aqueous NH(4)(+)CH(3)COO(-) (17:3, v/v) over a distance of 6 cm. Dequalinium chloride shows an R(F) value of 0.58. Pure dequalinium chloride is measured in the wavelength range from 200 to 500 nm and shows several by-products, contour plot visualized in absorption, fluorescence and using the Kubelka-Munk transformation. Scanning of a single track in absorption and fluorescence measuring 600 spectra in the range from 200 to 1100 nm takes 30s. As a sample pre-treatment of an ointment it is simply dissolved in methanol and can be quantified in absorption from 315 to 340 nm. The same separation can also be quantified using fluorescence spectrometry in the range from 355 to 370 nm. A new staining method for dequalinium chloride, using sodium tetraphenyl borate/HCl in water allows a fluorescence quantification in the range from 445 to 485 nm. The linearity range of absorption and fluorescence measurements is from 10 to 2000 ng. Sugar-containing preparations like liquids or lozenges with a reduced sample pre-treatment can be reliably quantified only in fluorescence. The total for the quantification of an ointment sample (measuring four standards and five samples), including all sample pre-treatment steps takes less than 45 min!
Enzyme‐assisted HPTLC method for the simultaneous analysis of inositol phosphates and phosphate
(2023)
Background
The analysis of myo‐inositol phosphates (InsPx) released by phytases during phytic acid degradation is challenging and time‐consuming, particularly in terms of sample preparation, isomer separation, and detection. However, a fast and robust analysis method is crucial when screening for phytases during protein engineering approaches, which result in a large number of samples, to ensure reliable identification of promising novel enzymes or target variants with improved characteristics, for example, pH range, thermal stability, and phosphate release kinetics.
Results
The simultaneous analysis of several InsPx (InsP1‐InsP4 and InsP5 + 6) as well as free phosphate was established on cellulose HPTLC plates using a buffered mobile phase. Inositol phosphates were subsequently stained using a novel enzyme‐assisted staining procedure. Immobilized InsPx were hydrolyzed by a phytase solution of Quantum® Blueliquid 5G followed by a molybdate reagent derivatization. Resulting blue zones were captured by DAD scan. The method shows good repeatability (intra‐day and intra‐lab) with maximum deviations of the Rf value of 0.01. The HPTLC method was applied to three commercially available phytases at two pH levels relevant to the gastrointestinal tract of poultry (pH 5.5 and pH 3.6) to observe their phytate degradation pattern and thus visualize their InsPx fingerprint.
Conclusion
This HPTLC method presents a semi‐high‐throughput analysis for the simultaneous analysis of phytic acid and the resulting lower inositol phosphates after its enzymatic hydrolysis and is also an effective tool to visualize the InsPx fingerprints and possible accumulations of inositol phosphates.
We report improved separation of the highly toxic contact herbicides paraquat, diquat, difenzoquat, mepiquat, and chloromequat by HPTLC. Quantification was based on a new derivatization reaction using sodium tetraphenylborate. Measurements were in the wavelength range from 440 to 480 nm or from 440 to 590 nm. An LED emitting very intense light at 365 nm was used for excitation. The quantification limits of paraquat and diquat in water, using improved solid-phase extraction, was in the low ng L −1 range. The linear range covered more than two orders of magnitude. Recovery was investigated for all the compounds, and was insufficient, ranging from 11 to 92%, but the method is inexpensive, rapid, and works reliably.
We present a planar chromatographic separation method for the compounds caffeine, artemisinin, and equol, separated on high-performance thin-layer chromatography (HPTLC) silica gel plates. As solvents for separation, methyl t-butyl ether and cyclohexane (1:1, V/V) have been used for equol, cyclohexane and ethyl acetate (7:3, V/V) for artemisinin, and ethyl acetate and acetone (7:3, V/V) for caffeine. After separation, the plate was scanned with a very specific time of flight-direct analysis in real time-mass spectrometry (TOF-DART-MS) system using the (M + 1)+ signals of equol, artemisinin, and caffeine. The (M + 1) peak of artemisinin at 283.13 m/z is clearly detectable, which is the proof that DART-MS is applicable for the quantitative determination of rather instable molecules. The planar set-up of DART source, HPTLC plate and detector inlet in a line showed higher sensitivities compared to desorption at an angle. The optimal detector voltage increases with the molar mass of the analyte, thus an individual determination of optimal detector voltage setting for the different analyte is recommended to achieve the best possible measurement conditions. In conclusion, DART-MS detection in combination with an HPTLC separation allows very specific quantification of all three compounds.