Detection of Hydroperoxides

Hydrogen peroxide and organic hydroperoxides can be reduced by molybdate-activated iodide [Allen et al. 1952 reactions (13) and (14)]. The iodine atoms formed in these reactions combine to iodine which complexes with iodide [reactions (15) and (16)], and it is this I3- complex which is measured (s(3540 nm) = 25,000 dm3 mol-1 cm-1) 

---

Analysis methods for the determination of hydroperoxides and hydroxyhydro-peroxides in reaction mixtures were developed and applied to photolysis mixtures of aliphatic and aromatic hydrocarbons, carbonyls, alcohols and halogenated hydrocarbons. HPLC with chemiluminescence, electrochemical and fluorescence detection was used for selective detection of hydroperoxides and hydroxohydro-peroxides and gas chromatography with mass spectrometric and flame ionisation detection for the analysis of all reaction products. 

The detection of hydroperoxides in this method is based on chemiluminescence. The system was optimised using r-buthylhydroperoxide, f-amylhydro-peroxide and cumenehydro-peroxide. Cytochrome C was used as a catalyst, the eluent was 0.25 M borate buffer. Detection limits were 5 nmol of active oxygen, which was only sufficient for laboratory experiments. 

Esser, G. Klockow, D. Detection of hydroperoxides in combustion aerosols by supercritical fluid extraction coupled to thin layer chromatography. Mikrochim. Acta 1994,113 (3-6), 373-379. 

Although the HPLC-CL approach allows the detection of hydroperoxides at the picomole levels in oxidized lipid extracts from complex biological samples, many problems arise in recovery of different hydroperoxides, in differences in substrate specificities between CL cocktails, mistaken structural assignment for different HPLC peaks, and spurious results resulting from some artificial... 

Frei, B., Yamamoto, Y., Nicolas, D. and Ames, B.N. Evaluation of an isoluminol chemiluminescence assay for the detection of hydroperoxides in human plasma. Anal. Biochem. 

As far as oxidation of the polymer with oxygen of the air is concerned, the /3-hydrogen atom in the neighborhood of the C=C double bond is the most likely one to be attacked by oxygen with the formation of hydroperoxide which undergoes further decomposition [19]. OH and CO groups have been detected spectroscopically in the polymer [67,83]. 

Many impurities are present in commercial caprolactam which pass into the liquid wastes from PCA manufacture from which caprolactam monomer may be recovered. Also, the products of die thermal degradation of PCA, dyes, lubricants, and other PCA fillers may be contained in the regenerated CL. Identification of die contaminants by IR spectroscopy has led to the detection of lower carboxylic acids, secondary amines, ketones, and esters. Aldehydes and hydroperoxides have been identified by polarography and thin-layer chromatography. 

A number of methods are available for following the oxidative behaviour of food samples. The consumption of oxygen and the ESR detection of radicals, either directly or indirectly by spin trapping, can be used to follow the initial steps during oxidation (Andersen and Skibsted, 2002). The formation of primary oxidation products, such as hydroperoxides and conjugated dienes, and secondary oxidation products (carbohydrides, carbonyl compounds and acids) in the case of lipid oxidation, can be quantified by several standard chemical and physical analytical methods (Armstrong, 1998 Horwitz, 2000). 

Detection of cholesteryl linoleate hydroperoxides and phosphatidylcholine hydroperoxides 63... 

The water (moisture) content can rapidly and accurately be determined in polymers such as PBT, PA6, PA4.6 and PC via coulometric titration, with detection limits of some 20 ppm. Water produced during heating of PET was determined by Karl Fischer titration [536]. The method can be used for determining very small quantities of water (10p,g-15mg). Certified water standards are available. Karl Fischer titrations are not universal. The method is not applicable in the presence of H2S, mercaptans, sulfides or appreciable amounts of hydroperoxides, and to any compound or mixture which partially reacts under the conditions of the test, to produce water [31]. Compounds that consume or release iodine under the analysis conditions interfere with the determination. 

In fact the extremely rapid reaction of NOH with hydroperoxides combined with the ready oxidation of hydroxylamines to nitroxides during storage even in the solid state makes unlikely the detection of >N0H from hindered amines in photo-oxidizing polymer. 

The formation and role of hydroperoxide groups, particularly in the early stages of polymer oxidation is well discussed in the introduction to the next chapter and also features in many of the references cited in this chapter. Their detection and quantification is therefore important. Although this can be done directly or implicitly through many of the instrumentation techniques discussed in this chapter, there are several tests that have been developed, some of which are still widely used, that are based more on chemical methods, titration or staining. The majority have been applied to polyolefins, especially polyethylene. 

POOH + 31 + 2H+ I3 + H20 + POH The method does not usually detect dialkylperoxides, POOP [16]. In applying the method, a known weight of sample will be refluxed in a mixture of isopropanol and acetic acid in the presence of sodium iodide. The reaction time is usually about 30 min. A detection limit of ca. 30 ppm of hydroperoxide has been reported [1]. 

A potentially powerful probe for sorting out the contribution of hydroperoxide-dependent and mixed-function oxidase-dependent polycyclic hydrocarbon oxidation is stereochemistry. Figure 9 summarizes the stereochemical differences in epoxidation of ( )-BP-7,8-dihydrodiol by hydroperoxide-dependent and mixed-function oxidase-dependent pathways (31,55,56). The (-)-enantiomer of BP-7,8-dihydrodiol is converted primarily to the (+)-anti-diol epoxide by both pathways whereas the (+)-enantiomer of BP-7,8-dihydrodiol is converted primarily to the (-)-anti-diol epoxide by hydroperoxide-dependent oxidation and to the (+)-syn-diol epoxide by mixed-function oxidases. The stereochemical course of oxidation by cytochrome P-450 isoenzymes was first elucidated for the methycholanthrene-inducible form but we have detected the same stereochemical profile using rat liver microsomes from control, phenobarbital-, or methyl-cholanthrene-induced animals (32). The only difference between the microsomal preparations is the rate of oxidation. 

Nourooz-Zadeh J. 1999. Ferrous ion oxidation in presence of xylenol orange for detection of lipid hydroperoxides in plasma. Methods Enzymol 300 58-62. 

Lipid Peroxidation Chemistry, 105, 273 overview of methods used for detecting lipid peroxidation, 105, 283 chemical methods for the detection of lipid hydroperoxides, 105, 293 comparative studies on different methods of malonaldehyde determination, 105, 299 concentrating ethane from breath to monitor lipid peroxidation in vivo, 105, 305. 

The following spectrophotometric methods are conceptually related to the POV however, they are intended for other purposes. HPLC-UVD at 234 nm was applied to detect lipid hydroperoxides, obtained in a set of experiments for assessing the effectiveness of lipoxygenase enzymes extracted from various microorganisms. The absorbance. A, measured at three different wavelengths is linearly correlated to the concentrations in xM units of lipid hydroperoxides, Clh, 7-oxocholesterol, Cqc and dienals, Cde according to the set of simultaneous equations 60. This method was used to track the Cu(II)-induced oxidation of LDL" °. 

RP-HPLC with nonaqueous solvents and UVD at 246 nm was developed for the determination of low level POVs of vegetable oils. These measurements are specific for conjugated diene peroxides derived from vegetable oils with relatively high linoleic acid content. These measurements may be supplemented by nonspecific UVD at 210 nm and ELSD for detection of all eluted species. The elution sequence of the triglycerides in a nonaqueous RP-HPLC is linearly dependent on the partition number of each species, Vp, which is defined as = Nq — 2Ni, where Nq is the carbon number and is the double bond number. In the case of hydroperoxides = Nq — 2Nd — Vhpo, where Vhpo is the number of hydroperoxyl groups in the molecule (usually 1 for incipient POV). For... 

Hydroperoxides may be determined by measuring at 290 nm (e = 44100 M cm ) or 360 nm (e = 28000 cm ) the concentration of 13 formed in the presence of a large excess of ions. The reaction may be too slow for practical purposes, unless a catalyst is present. For example, an assay for lipid hydroperoxides conducted without a catalyst may require several measurements every 6 min until the absorbance reaches a maximum. Exclusion of air from the sample cuvette is important. The method is about 1000-fold more sensitive than thiosulfate titration The iodometric method with UVD at 360 was adopted for detecting the presence of hydroperoxides derived from protein, peptide or amino acid substrates subjected to y-radiation, after destroying the generated H2O2 with catalase. ... 

A procedure for determination of lipid hydroperoxides in human plasma is based on kinetic measurement of the CL of luminol (124) with hemin (75a) catalysis . CLD of microperoxidase-catalyzed oxidation of luminol (124) or isoluminol (190) was applied to detection and determination of amino acid hydroperoxides after exposure to UV and y-irradiation A method for determination of hydroperoxides in the phospholipids of cultured cells uses isoluminol (190) and microperoxidase as catalyst " . Simultaneous determination of phosphatidylcholine hydroperoxides and cholesteryl ester hydroperoxides in human serum is carried out by quantitative extraction of the lipids, HPLC separation by column switching and CLD using isoluminol (190) with microperoxidase catalysis . ... 

---