Quantitation HPLC

Quantitative HPLC analysis was carried out on a Spectraphysics 8720 chromatography system, a rapid scan detector by Barspec on a Zorbax ODS column with acetonitrile water 75/25 as the eluent. 

A sample of HBCD was weighed in a glass tube dipped in an oil bath with a temperature regulator of 1°C. During the experiment the sample was monitored for HBr formation. At different times the samples were taken out and cooled to room temperature, weighed and analyzed by quantitative HPLC. 

The structure of the last eluting peak (Fig. ID, peak 10), structure is easily deduced to be 2,2, 3,3, 4,4, 5,5, 6,6 decabromo DPF from the bromine content of the pure sample, lack of proton spectrum and long retention time. Indeed, a recrystallized standard serves as reference material for a quantitative HPLC assay to be described elsewhere. 

D. Cristea, I. Bareau and G.Vilarem, Identification and quantitative HPLC analysis of the main flavonoids present in weld (Reseda luteola L.), Dyes and Pigments, 57, 267 272 (2003). 

One of the best tools for metabolite profiling is the hybrid QTRAP MS/MS system (Applied Biosystems).119-121 While the hybrid QTRAP MS/MS was initially considered a premier tool for metabolite identification, it has more recently been seen as a tool for quantitation and metabolite profiling. Li et al.122 described the use of a hybrid QTRAP MS/MS system for discovery PK assays plus metabolite profiling in the same analytical procedure. Because QTRAP MS/MS may be used as a triple quadrupole MS system, it can be used as part of a quantitative HPLC/MS/MS system. Because QTRAP MS/MS also has linear ion trap capabilities, it can be used for metabolite screening and characterization—essentially it combines the capabilities of a triple quadrupole mass spectrometer and a linear ion trap mass spectrometer. 

Product predictions for nucleophilic additions to C q, based on AMI calculations, show that among the many possible isomers a few are energetically favored [29]. Two areas within the molecule can be distinguished, which are the inert belt at the equator and the more reactive CgQ-like double bonds at the poles (Figure 3.4). Experimentally, hydroalkylation and hydroarylation reactions of C q under quantitative HPLC control yield predominantly one isomer of each C7oHR[4]. 

Hertog, M.G.L., Hollman, P.C.H., and Venema, D.P., Optimization of a quantitative HPLC determination of potentially anticarcinogenic flavonoids in vegetables and fruits, J. Agric. Food Chem., 40, 1591, 1992. 

Shackleton CH, Reid S (1989) Diagnosis of recessive X-linked ichthyosis quantitative HPLC/ mass spectrometric analysis of plasma for cholesterol sulfate. Clin Chem 35 1906-1910... 

We thank Dr. John R. Porter for advice on plant tissue culture and phytochemical methods and for critical reading of the manuscript. Dr. Anil D Mello for his advice concerning the development of quantitative HPLC methodology, and Jeremy Mihalov, Anne Giordano and Mila Denisova for their invaluable contributions. We gratefully acknowledge PCPS for financial support, namely the Evelyn C. 

Semipreparative HPLC has been employed to obtain a vitamin D-rich fraction of the unsaponifiable matter for subsequent quantitative HPLC. Combinations of chromatographic modes used for offline semipreparative and quantitative analysis have included polar bonded-phase/adsorption (211,212), reversed-phase/adsorption (194,213), and adsorption/reversed-phase (70,125). An online two-dimensional HPLC technique using two polar bonded-phase columns has also been described (214). 

Food Sample preparation Semipreparative HPLC Quantitative HPLC Compounds separated Ref. 

A qualitative and quantitative HPLC method for analysis of mixtures of 12 antioxidants was described Grosset et al. (121). For the identification of the components present, gradient elution with a convex profile from 35 65 water-methanol to pure methanol is used, on a Waters 5-/xm C18 column, with UV detector. Propyl gallate was not separated by this system. For quantitative analysis, with UV and electrochemical detectors in series, the water-methanol mixture or pure methanol was used as the eluent, under isocratic conditions, with lithium perchlorate as supporting electrolyte. An applied potential ranging from +0.8 to +1.7 V allows detection of all the antioxidants tested. Both modes of detection were very sensitive, with limits of detection as low as 61 pg. 

Matuszewski, B. K. (2006). Standard hne slopes as a measure of a relative matrix effect in quantitative HPLC-MS bioanalysis. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 830 293-300. 

A quantitative HPLC method for the analysis of sphingolipids as their perbenzoyl derivatives was first developed for ceramides (5). Ceramides can be conveniently derivatized with benzoic anhydride in pyridine (3 hrs at 110°C) and the products formed have been utilized for the quantitative analysis of NFA and HFA ceramides in normal and Farber s disease tissue. Iwamori and Moser also utilized this procedure for the analysis of ceramides in Farber s disease urine (6). More recently Iwamori and Moser (7) established that the ceramide derivatives formed by reaction with benzoyl chloride or benzoic anhydride are analogous to those formed with cerebrosides. They also characterized the behavior... 

The content of neutral glycolipids in the fractions of interest was determined by the quantitative HPLC method of Ullman and McCluer (3). 

Ikeda Y, Fujii Y, Nakaya I and Yamazaki M (1995) Quantitative HPLC analysis of cardiac glycosides in Digitalis purpurea leaves. J Nat Prod 58, 897-901. 

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