HPLC-diode array detection

An HPLC method for chlorogenic acids with lactones in six different commercial brands of roasted coffee was developed by Schrader et al. (143). Hydroxycinnamic acid derivatives, including mono- and di-caffeoylquinic acids, corresponding lactones, and feruloylquinic acids were extracted from coffee with methanol at 80°C for 1 h under reflux. An HPLC method using step-gradient elution with 2% aqueous acetic acid (eluent A) and ACN (eluent B) for a 75-min run time was developed. Determination was carried out by HPLC with UV detection at 324 nm, and further confirmation was conducted by HPLC-thermospray (TSP)-MS and HPLC-diode array detection. Elution order for mono-caffeoylquinic acid (CQA) was 3-CQA, 5-CQA, followed by 4-CQA, which was different from the usual elution order of mono-CQA (Fig. 17). These results indicate that it is currently not possible to predict the elution order of different reversed-phase packings due to the different selectivity (143). 

PB Andrade, ARF Carvalho, RM Seabra, MA Ferreira. A previous study of phenolic profiles of quince, pear, and apple purees by HPLC diode array detection for the evaluation of quince puree genuineness. J Agric Food Chem 46 968-972, 1998. 

PB Andrade, R Leitao, RM Seabra, MB Oliveira, MA Ferreira. Development of an HPLC/diode-array detection method for simultaneous determination of seven hydroxycinnamic acids in green coffee. J Liquid Chromatogr Related Technol 20 2023-2030, 1997. 

Method and equipment solid phase extraction and high performance liquid chromatography (HPLC)/diode array detection. 

S. C. Rastogi, V. J. Barwick, and S. V. Carter, Identification of organic colorants in cosmetics by HPLC-diode array detection, Chromatographia 45 215 -228 (1997). 

Saleh, M.H. and Tan, B. 1988. HPLC-diode array detection technique for the identification and quantification of carotene formulations. Anal. Biochem. 33 247-252. 

Important applications of Af-PLS are in the area of multivariate calibrations for excitation/emission fluorescence spectrometry, for hyphenated analytical methods, such as HPLC/diode array detection and GC/MS, or for multidimensional separation techniques with or without coupling to spectroscopy. 

Identification Backpropagation HPLC diode array detection [17]... 

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.

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

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. 

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. 

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... 

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]. 

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). 

For the characterisation of the biodegradation intermediates of C12-LAS, metabolised in pure culture by an a-proteobacterium, Cook and co-workers [23] used matrix-assisted laser desorption/ionisation (MALDI)-time of flight (TOF)-MS as a complementary tool to HPLC with diode array detection and 1H-nuclear magnetic resonance. The dominating signal in the spectrum at m/z 271 and 293 were assigned to the ions [M - H] and [M - 2H + Na]- of C6-SPC. Of minor intensity were the ions with m/z 285 and 299, interpreted to be the deprotonated molecular ions of C7- and C8-SPC, respectively. 

Numerous CE separations have been published for synthetic colours, sweeteners and preservatives (Frazier et al., 2000a Sadecka and Polonsky, 2000 Frazier et al., 2000b). A rapid CZE separation with diode array detection for six common synthetic food dyes in beverages, jellies and symps was described by Perez-Urquiza and Beltran (2000). Kuo et al. (1998) separated eight colours within 10 minutes using a pH 9.5 borax-NaOH buffer containing 5 mM /3-cyclodextrin. This latter method was suitable for separation of synthetic food colours in ice-cream bars and fmit soda drinks with very limited sample preparation. However the procedure was not validated for quantitative analysis. A review of natural colours and pigments analysis was made by Watanabe and Terabe (2000). Da Costa et al. (2000) reviewed the analysis of anthocyanin colours by CE and HPLC but concluded that the latter technique is more robust and applicable to complex sample types. Caramel type IV in soft drinks was identified and quantified by CE (Royle et al., 1998). 

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