Diode-array detection

In contrast, diode-array scanners use white light for illumination pmposes. Diode-array scanners register absorption and fluorescence spectra and densitograms at different wavelengths simultaneously from the plate. This kind of scanner was first introduced in 2000 by Spangenberg [84b]. 

Another 25 fibers in slit arrangement illuminate the plate for fluorescent measurements by use of an LED, emitting dense light at 365 nm. A reduced attachment for absorption measurements in the UV-range consists of two 25 fibber arrays each, to iUu-minate the plate by use of a deuterium lamp and to transport the scattered hght back to the diode-array detector. 

For evaluation purposes, the reflected light from a clean TLC-plate (Jo) is combined with the reflected light of a TLC-track (J). The combination of the wavelength dependent light distributions are combined according to expression (1). 

To scan an absorption spectrum directly from a sample spot, a mono-wavelength scanner has to illuminate the plate with light of different wavelengths. Using a diode-array scaimer the absorption spectrum of a TLC-plate spot can be directly extracted from the measured scan. 

The well-known Kubelka-Munk equation (4) is the combination of equation (2) and equation (3) and describes both, absorption and fluorescence spectra [84c]. 

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A powerful tool now employed is that of diode array detection (DAD). This function allows peaks detected by UV to be scanned, and provides a spectral profile for each suspected microcystin. Microcystins have characteristic absorption profiles in the wavelength range 200-300 nm, and these can be used as an indication of identity without the concomitant use of purified microcystin standards for all variants. A HPLC-DAD analytical method has also been devised for measurement of intracellular and extracellular microcystins in water samples containing cyanobacteria. This method involves filtration of the cyanobacteria from the water sample. The cyanobacterial cells present on the filter are extracted with methanol and analysed by HPLC. The filtered water is subjected to solid-phase clean-up using C g cartridges, before elution with methanol and then HPLC analysis. 

Figure 5.3 Analysis of 100 ml of (a) surface water and (b) drinking water sample spiked with 0.1 pig/ml of microcystins, using column-switching HPLC 1, microcystin-RR 2, microcystin-YR 3, microcystin-LR. Reprinted from Journal of Chromatography A, 848, H. S. Lee et al, On-line trace enrichment for the simultaneous determination of microcystins in aqueous samples using high performance liquid chromatography with diode-array detection , pp 179-184, copyright 1999, with permission from Elsevier Science.

C. Aguilar, I. Feirer, R Bonnll, R. M. Marce and D. Barcelo, Monitoring of pesticides in river water based on samples previously stored in polymeric cartridges followed by on-line solid-phase extraction-liquid cliromatography-diode array detection and confirmation by atmospheric pressure chemical ionization mass spectrometry . Anal. Chim. Acta 386 237-248 (1999). 

R. M. Marce, H. Prosen, C. Crespo, M. Calull, R Boirull and U. A. Th Brinkman, Online ti ace enrichment of polar pesticides in environmental waters by reversed-phase liquid cliromatography-diode array detection-particle beam mass spectrometry , J. Chromatogr. 696 63-74 (1995). 

S. Lacorte, J. J. Vreuls, J. S. Salau, R Ventura and D. Barcelo, Monitoring of pesticides in river water using fully automated on-line solid-phase extraction and liquid cliro-matography with diode array detection with a novel filrtation device , J. Chromatogr. 795 71-82(1998). 

C. Aguilar, R Bomtll and R. M. Marce, Determination of pesticides by on-line trace enrichment-reversed-phase liquid cliromatogr aphy-diode-array detection and confirmation by particle-beam mass spectrometry , Chromatographia 43 592-598 (1996). 

I. Eeirer, V. Pichon, M. C. Hennion and D. Barcelo, Automated sample preparation with exti action columns by means of anti-isoproturon immunosorbents for the determination of phenylurea herbicides in water followed by liquid chi omatography diode array detection and liquid clrromatogi aphy-atmospheric pressure chemical ionization mass spectrometry , 7. Chromatogr. Ill 91-98 (1997). 

Determined by HPLC using diode array detection of a sample taken directly from the reaction mixture. b (< = 1, CHClj). 

Keller, H. R. and Massart, D. L., Artefacts in Evolving Factor Analysis-Based Methods for Purity Control in Liquid Chromatography with Diode-Array Detection, Ana/yt/ca Chimica Acta 263, 1992, 21-28. 

MATTILA p and KUMPULAINEN J (2002) Determination of free and total phenolic acids in plant-derived foods by HPLC with diode-array detection, JAgric Food Chem, 50, 3660-67. 

Cortes et al.. Identification and quantification of carotenoids including geometrical isomers in fruit and vegetable juices by liquid chromatography with ultraviolet-diode array detection, J. Agric. Food Chem., 52, 2203, 2004. 

Hvattum, E., Determination of phenolic compounds in rose hip (Rosa canina) using liquid chromatography coupled to electrospray ionisation tandem mass spectrometry and diode-array detection, Rapid Commun. Mass Spectrom., 16, 655, 2002. 

Saito, K., A new enzymatic method for extraction of precarthamin from dyer s saffron (Carthamus tinctorius) florets, Z. Lebensmitt. Untersuch. Forsch., 197, 34, 1993. Cserhati, T. et ah. Separation and quantitation of colour pigments of chili powder (Capsicum frutescens) by high-performance liquid chromatography-diode array detection, J. Chromatogr. A, 896, 69, 2000. 

Spectrophotometric resolution for the discrimination of individual colorant molecules found in mixtures is lower than that of chromatographic techniques such as TLC or HPTLC and even low-cost paper chromatography. More expensive but more accurate determinations may be made by RP-HPLC, IP-HPLC with UV-Vis, and diode array detection. ... 

Recently a new method was developed for the complete liquid chromatographic separation and diode array detection of standard mixtures of the 14 most frequently used synthetic colorants. Protocols for RP-HPLC - " and IP-HPLC techniques have been extensively described and the techniques were compared with micellar electrokinetic capillary chromatography, - which has been shown to be suitable for the analysis of synthetic colorants. 

Five synthetic and five natural colorants were identified and quantified in lyo-philized dairy products and fatty foods using an automatic method based on solid phase extraction using a stationary phase followed by RP-HPLC C,g columns for the sequential retention of colorants and diode array detection. Lyophilization of the samples coupled with the separation procedure provided clean extracts despite the complexity of the food matrices and preserved the sample for at least 2 months without changes in colorant concentrations. The detection limits achieved for the colorants were found in a wide range from 0.03 to 75 pg/g of the lyophilized sample, according to the limits established by the European Union. ... 

Diode Array Detection in HPLC, edited by Ludwig Huber and Stephan A. George... 

B.G.M. Vandeginste, R. Essers, T, Bosman, J. Reijnen and G. Kateman, Three-component curve resolution in HPLC with multi wavelength diode array detection. Anal. Chem., 57 (1985) 971-985. 

H.R. Keller and D.L. Massart, Artifacts in evolving factor analysis-based methods for peak purity control in liquid-chromatography with diode array detection. Anal. Chim. Acta, 263 (1992) 21-28. 

J. Chen and S.C. Rutan, Identification and quantification of overlapped peaks in liquid chromatography with UV diode array detection using an adaptive Kalman filter. Anal. Chim. Acta, 335(1996) 1-10. 

Polymeric precolumns of styrene-divinylbenzene were used by Aguilar et al. to monitor pesticides in river water. Water samples (50 mL) were trace enriched on-line followed by analysis using LC combined with diode-array detection. LC atmospheric pressure chemical ionization (APCI) MS was used for confirmatory purposes. It was found that after the pesticides had been extracted from the water sample, they could be stored on the precartridges for up to 3 months without any detectable degradation. This work illustrates an advantage of SPE for water samples. Many pesticides which may not be stable when stored in water, even at low temperature, may be extracted and/or enriched on SPE media and stored under freezer conditions with no detectable degradation. This provides an excellent way to store samples for later analysis. 

Dong, M. W. and Greenberg, A., Liquid chromatographic analysis of polynuclear aromatic hydrocarbons with diode array detection, /. Liq. Chromatogr., 11, 1887, 1988. 

A polyethylene-coated (PEE) silica column was used with water-methanol eluents to achieve the separation and retention of 27 pesticides.40 The retention times of 33 commercial pesticides were determined on an octadecyl (ODS)-derivatized alumina column using water-methanol eluents and compared with retention properties on an ODS-silica column packing.41 More recently, RP-HPLC was used in combination with diode array detection for the identification and quantification of 77 pesticides (acidic, basic, and neutral) in groundwater samples.42... 

For many applications, diode array detection has become routine. A photodiode array was used for simultaneous detection of 100 capillaries in zone electrophoresis and micellar electrokinetic chromatography (MEKC).1516 Deflection of a laser beam by acoustic waves was reported as a means to scan six capillary channels on a microchip.17 The design of a low-noise amperometric detector for capillary electrophoresis has been reported.18... 

Drushel [58] and others [31,59] have described the needs of the chromatographer in the area of detectors. Specific texts concern detection in quantitative GC [54], diode-array detection in HPLC [48], selective detectors [39] and element-specific chromatographic detection by AES [60], electrochemical detectors [61] and laser detectors [62]. 

L. Huber and S. George, Diode-array Detection in High-performance Liquid Chromatography, M. Dekker, New York, NY (1993). 

Mei, D.A., Gross, G.J., and Nithipatikom, K., Simultaneous determination of adenosine, inosine, hypoxanthine, xanthine and uric acid in microdialysis samples using microbore column liquid chromatography with diode array detection, Analyt. Biochem., 238,34,1996. 

The most commonly used method for the identification of carotenoids is high-pressure liquid chromatography (HPFC) combined with the UV-Vis absorption detection. The introduction of diode array detection enabled parallel collection of pigment spectra, which greatly aids the quantification and localization of unknown compounds. Coupling HPFC with the mass-spectrometer significantly... 

The iodine-catalyzed photoisomerization of all-trans- a- and (3-carotenes in hexane solutions produced by illumination with 20 W fluorescence light (2000 lux) and monitored by HPLC with diode-array detection yielded a different isomer distribution (Chen et al. 1994). Four cis isomers of [3-carotene (9-cis, 13-cis, 15-cis, and 13,15-cli-r/.v) and three cis isomers of a-carotene (9-cis, 13-cis, and 15-ri.v) were separated and detected. The kinetic data fit into a reversible first-order model. The major isomers formed during the photosensitized reaction of each carotenoid were 13,15-di-d.v- 3-carotene and 13-ds-a-carotene (Chen et al. 1994). 

LC-MS-MS was also the method of choice for the analysis of UV filters in solid matrices. Both LC and UPLC have been applied in three out of the four methods available for the determination of UV filters in sludge. Separation was performed on C8 and C18 LC-chromatographic columns (Zorbax, Eclipse, Vydac, and Purosphere) using binary gradient elution of mobile phases consisting of water/ methanol or water/acetonitrile. MS-MS detection was performed in SRM with ESI and atmospheric pressure photoionization (APPI) in both positive and negative modes. For each compound, two characteristics transitions were monitored. In addition to MS and MS-MS, diode array detection (DAD) was occasionally applied to the determination of OT. Spectra were recorded between 240 and 360 nm and discrete channels at 310 nm. 

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