Reversed-phase high performance liquid quantification

Endo, Y., Tagiri-Endo, M., Seo, H. S., and Fujimoto, K. (2001). Identification and quantification of molecular species of diacylglyceryl ether by reversed-phase high-performance liquid chromatography with refractive index detection and mass spectrometry. J. Chro-matogr. A 911, 39-45. 

Lim CK, Peters TJ (1984) Urine and faecal porphyrin profiles by reversed-phase high-performance liquid chromatography in the porphyrias. Clin Chim Acta 139 55-63 Minder El, Vuilleumier JP, Vonderschmitt DJ (1992) Prototype application of robot in the clinical laboratory enabling fully automated quantification of fecal porphyrins. Clin Chem 38 516-521... 

A reversed-phase high performance liquid chromatographic method for the determination of salvinorin A, a psychotropic diterpene isolated from the Mexican sage Salvia divinorum, has been developed. Extracts from several plant collections were examined on a C-18 column with UV detection and isocratic elution with acetonitrile water (45 55). This assay allowed quantification of salvinorin A in extracts of leaves and stems of A , divinorum and has also been applied to the screening of related species for the production of salvinorin A. Levels of salvinorin A in leaves range from 0.89 to 3.70 mg/g dry weight. 1999 John Wiley Sons, Ltd. 

In this unit, methods for reversed-phase high-performance liquid chromatography (HPLC) are described for the analysis of polyphenolics. HPLC analysis can be employed in an easy and fast manner to obtain an accurate elucidation and quantification of individual polyphenolic compounds found in plant-based materials. The separation of each polyphenolic is based on the polarity differences among polyphenolics with structural similarities and uses various combinations of mobile and stationary phases. 

Veloso, A. C. A., Teixeira, N., and Ferreira, I. M. P. L. V. O. (2002). Separation and quantification of the major casein fractions by reverse-phase high performance liquid chromatography and urea-polyacrylamide gel electrophoresis Detection of milk adulterations.. Chromatogr. A 967, 209-218. 

Mantoura and Llewelyn [3] developed a reversed phase high performance liquid chromatographic system for a rapid (about 20min) separation and quantification of 14... 

Atta-Politou, J. Parissi-Poulou, M. Dona, A. Koutselinis, A. A simple and rapid reversed phase high performance liquid chromatographic method for quantification of alprazolam and ot- hydroxyalpra-zolam in plasma. J.Liq.Chromatogr., 1991,14, 3531-3546 [plasma LOD 1 ng/mL extracted hydrox-yalprazolam flunitrazepam (IS)]... 

Gutenberger, S.K. Olson, D.P. Kagel, R.A. Comparison of reversed phase high-performance liquid chromatography and competitive protein binding assay in the quantification of cortisol in bovine plasma. J.Liq.Chromatogr., 1985, 8, 107-124 [plasma cow dexamethasone (IS)]... 

Hartwick, R.A., Assenza, S.P. and Brown, P.R., 1979. Identification and quantification of nucleosides, bases and other UV absorbing compounds in serum using reversed phase high performance liquid chromatography. I. Chromatographic methodology. In A. Ziatkis (Editor), Advances in Chromatography. Lausanne Symp., pp. 725—736. 

Sharma U, Sharma K, Sharma N, Sharma S, Singh HP, Sinha AK, Microwave-assisted efficient extraction of different parts of hippophae rhamnoides for the comparative evaluation of antioxidant activity and quantification of its phenolic constituents by reverse-phase high-performance liquid chromatography (RPHPLC), J. Agric. Food Chem. 2008 56 374-379. 

Bobe, G. Beitz, D.C. Freeman, A.E. Lindberg, G.L. Separation and quantification of bovine milk proteins by reversed-phase high-performance liquid chromatography. J. Agric. Food Chem. 1998, 46, 458-463. 

Elgar, D.F. Norris, C.S. Ayers, J.S. Pritchard, M. Otter, D.E. Palmano, K.P. Simultaneous separation and quantification of the major bovine whey proteins including proteose peptone and caseinomacropeptide by reversed-phase high-performance liquid chromatography on polystyrene ivi-nylbenzene. J. Chromatogr. A, 2000, 878, 183-196. 

Terao, J., Shibata, S.S. and Matsushita, S. Selective quantification of arachidonic acid hydroperoxides and their hydroxy derivatives by reverse-phase high-performance liquid chromatography. Anal. Biochem. 169,415—423 (1988). 

Catena, E., Perez, G., Sadaba,B.,Azanza,J.R., Campanero, M.A. (2009) A fast reverse-phase high performance liquid chromatographic tandem mass spectrometry assay for the quantification of clindamycin in plasma and saliva using a rapid resolution package. Journal of Pharmaceutical and Biomedical Analysis, 50, 649-654. 

Lebiedzinska, A., MarszaH, M., Kuta, J., and Szefer, P., 2007. Reversed-phase high-performance liquid chromatography method with coulometric electrochemical and ultraviolet detection for the quantification of vitamins Bj (thiamine), Bg (pyridoxamine, pyridoxal and pyridoxine) and B12 in animal and plant foods. Journal of Chromatography A. 1173 71 80. 

S Li, AB Barua, CA Huselton. Quantification of retinoyl-beta-glucuronides in rat urine by reversed-phase high-performance liquid chromatography with ultraviolet detection. J Chromatogr B Biomed Appl 683 155-162, 1996. 

Identification and quantification of natural dyes need high performance analytical techniques, appropriate for the analysis of materials of complicated matrices containing a small amount of coloured substances. This requirement perfectly fits coupling of modern separation modules (usually high performance liquid chromatography in reversed phase mode, RPLC, but also capillary electrophoresis, CE) with selective detection units (mainly mass spectrometer). 

Currently, high-performance liquid chromatography (HPLC) methods have been widely used in the analysis of tocopherols and tocotrienols in food and nutrition areas. Each form of tocopherol and tocotrienol can be separated and quantified individually using HPLC with either a UV or fluorescence detector. The interferences are largely reduced after separation by HPLC. Therefore, the sensitivity and specificity of HPLC methods are much higher than those obtained with the colorimetric, polarimetric, and GC methods. Also, sample preparation in the HPLC methods is simpler and more efficiently duplicated than in the older methods. Many HPLC methods for the quantification of tocopherols and tocotrienols in various foods and biological samples have been reported. Method number 992.03 of the AOAC International Official Methods of Analysis provides an HPLC method to determine vitamin E in milk-based infant formula. It could probably be said that HPLC methods have become dominant in the analysis of tocopherols and tocotrienols. Therefore, the analytical protocols for tocopherols and tocotrienols in this unit are focused on HPLC methods. Normal and reversed-phase HPLC methods are discussed in the separation and quantification of tocopherols and tocotrienols (see Basic Protocol). Sample... 

Gibson, C.R. et al. Electrospray ionization mass spectrometry coupled to reversed-phase ion-pair high-performance liquid chromatography for quantification of sodium boroc-aptate and application to pharmacokinetic analysis. Anal. Chem. 2002, 74, 2394—2399. 

Liquid chromatography is a separation technique which is used widely in many different areas of analytical chemistry and provides a powerful tool for the separation and quantification of substances in various matrices. Nowadays, the majority of the high-performance liquid chromatographic methods are carried out in reversed-phase mode using a nonpolar stationary phase and a polar mobile phase. 

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