Reagent Luminol

Basic procedure (ACL kit) Mix 2400 pL of ACL reagent 1 (diluter) with 100 pL of ACL reagent 2 (buffer) and 25 pL of photosensitizer reagent (luminol based). Start measurement after brief vortexing. Assayed solution (lipid extract) is added before addition of photosensitizer reagent. Volume of ACL reagent 1 is reduced by the volume of assayed solution. Standard substance a-tocopherol or Trolox. Duration of measurement 1 min. Measured parameter integral (area under the kinetic curve of PCL). 

Hool and Nieman succeeded in immobilizing the reagent (luminol) on a support (CPG) located in the flow-cell of a chemiluminescence detector. For solubility reasons, the reaction between luminol and glutaraldehyde was... 

Reagents luminol (5-amino-2,3-dihydro-1,4-phthalazinedione), horseradish... 

FIA-chemiluminescence sensors were made by immobilization of analytical reagents (luminol and ferricyanide, or luminol and permanganate) on an anion exchange resin column [75,86],... 

Carrier 0.1 M carbonate buffer, pH 10 CL reagents luminol/ microperoxidase in 0.1 M carbonate buffer, pH 10... 

Carrier methanol/ chloroform (9 1) mixture CL reagents luminol/ cytochrome c in borate buffer containing Triton X-100 (biphasic system) Carrier 100% methanol CL reagents lucigenin in methanol/borate buffer (4 1) (monophasic system)... 

Chemiluminescence and bioluminescence are also used in immunoassays to detect conventional enzyme labels (eg, alkaline phosphatase, P-galactosidase, glucose oxidase, glucose 6-phosphate dehydrogenase, horseradish peroxidase, microperoxidase, xanthine oxidase). The enhanced chemiluminescence assay for horseradish peroxidase (luminol-peroxide-4-iodophenol detection reagent) and various chemiluminescence adamantyl 1,2-dioxetane aryl phosphate substrates, eg, (11) and (15) for alkaline phosphatase labels are in routine use in immunoassay analyzers and in Western blotting kits (261—266). 

We have shown that known reaction of luminol with peroxydisulphate at low luminol concentrations takes place in the regime of controlled generation of SO ion-radicals at spontaneous destruction of peroxydisulphate. The detection limit for various types of antioxidants in water using this reaction is varied from 10 to 10 M. It is possible also to determine some polluting admixtures present in the atmosphere. The reagent used is the mixture of the luminol, base and K S O, which, once prepai ed, could be used during a working day. 

Calibration with luminol luminescence (Lee et al., 1966). Calibration with luminol can be performed in aqueous solution in the presence of H2O2 and a suitable catalyst (luminescence range 380-550 nm, Amax 430 nm), or in DMSO in the presence of potassium t-butoxide (A.mav 486 nm). The calibration method in aqueous medium requires the following three reagents ... 

Equipment and reagents Ultrasound generator, pulse generator, photomultiplier tube, oscilloscope, light-insulated cabinet, rare gas source (e.g. argon), 3-ami-nophthalhydrazide (luminol), sodium hydroxide, alcohol or other volatile organic solutes. 

Luminol amidine 132, synthesized from luminol and the Vilsmeier reagent from DMF and thionyl chloride, has been proposed as a suitable luminol derivative for analytical purposes because, unlike luminol, it can be easily purified by recrystallization from water. 132 exhibits a chemiluminescence quantum yield of about 20% of luminol in ferricyanide-catalyzed oxidation by aqueous alkaline hydrogen peroxide Amax of the emission is 452 nm 196>. 

Advances have been achieved in recent years, such as the use of CL reagents as labels to derivatize and sensitively determine analytes containing amine, carboxyl, hydroxy, thiol, and other functional groups and their application in HPLC and CE [35, 36], the synthesis and application of new acridinium esters [37], the development of enhanced CL detection of horseradish peroxidase (HRP) labels [38], the use of immobilization techniques for developing CL-based sensors [39-42], some developments of luminol-based CL in relation to its application to time-resolved or solid-surface analysis [43], and the analytical application of electrogenerated CL (ECL) [44-47], among others. 

Figure 10 shows the instrumental setup used to implement the APP-CLS approach. It consists of (a) a CSTR that is a thermostated 10-mL glass reaction vessel accommodated in a commercially available spectrofluorimeter (a Hitachi F2000 model in this case) (b) a four-channel peristaltic pump with three channels used to dispense the reagent solutions and the fourth to keep the volume of the reaction mixture in the CSTR constant the three reagent solutions are as follows (1) 0.15 M hydrogen peroxide (2) 0.15 M sodium thiocyanate, 0.15 M sodium hydroxide, and 1.95 x 10 3 M luminol and (3) 6.0 x 10 4 M copper(II) sulfate ... 

Some luminol derivatives have been developed as CL labeling reagents. Analytes prelabeled with luminol derivatives are separated by HPLC, mixed with postcolumn reagents such as hydrogen peroxide and an alkaline solution of potassium hexacyanoferrate (III), and then detected by a CL detector. Highly sensitive determination is possible by optimizing the conditions to increase the CL reaction efficiency for each analyte. 

Figure 7 (A) Luminol-type CL reagents and (B) derivatization reactions for DPH and...

Table 1 Application of Luminol Derivatives as CL Labeling Reagents to HPLC... 

Another variation on the luminol CL sensor for N02 was introduced by Collins and Ross-Pehrsson [12] where a solid-phase reagent was positioned below a PMT, across which the air under test is pumped. Of the hydrogel or polymeric sorbents investigated, a Waterlock superabsorbing polymer (hydrogel)... 

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