Deproteinized

A polymethacrylate copolymer is modified by successive reaction with epichlorohydrin, w-aminophenylboric acid, and nitric acid to introduce a 1-amino-(2 -nitrophenyl-5 -boric acid)-2-hydroxyl-3-o-propyl group. The modified polymethacrylates are used as chromatographic support materials and can be used to analyze biological materials without prior deproteinization (35). 

Drugs and Related Compounds Ampicillin human plasma deproteination C18 C18 post-Elu 28... 

Gianesello et al. (120) described the determination of the bronchodilator brox-aterol in plasma by on-line LC-GC. After deproteination and extraction, the LC separation was carried out by using a mixture of -pentane and diethyl ether (55 45 (vol/vol) as mobile phase. A small cut of the LC chromatogram (shown in Figure 11.9(a)) was introduced at 85 °C into the GC via so-called concurrent solvent evaporation. Figure 11.9(b) demonstrates that a detection limit of about 0.03 ng/ml was obtained. A fully automated LC-GC instrument was described by Munari and Grob (121) and its applicability was demonstrated by the determination of heroin metabo-... 

Owing to the complexity of multi-residue methods for products of animal origin, it is not possible to outline a simple scheme however, readers should refer to methods described in two references for detailed guidance (Analytical Methods for Pesticides in Foodstuffs, Dutch method collection and European Norm EN 1528. ) There is no multi-method specifically designed for body fluids and tissues. The latter matrix can be partly covered by methods for products of animal origin. However, an approach published by Frenzel et al may be helpful (method principle whole blood is hemolyzed and then deproteinized. After extraction of the supernatant, the a.i. is determined by GC/MS. The LOQ is in the range 30-200 ag depending on the a.i.). 

Turner and Warnock [56] determined miconazole in human saliva using high performance liquid chromatography. Deproteinated human saliva samples containing miconazole was chromatographed on a C8 reversed-phase radial compression column using 77% methanol in 0.01 M EDTA with 0.005 M w-nonylamine at a flow... 

Polymer Labs. PLRP-S 0.01M oxalic acid-ACN (75 25, v/v) UV 360 nm Animal tissues Extraction with oxalic buffer followed by chelation and deproteination, cleanup with styrene-divinylbenzene cartridge Rec 76-87% [77]... 

Rabenstein and Yamashita [52] determined penicillamine and its symmetrical and mixed disulfides by HPLC in biological fluids. Plasma and urine were deproteinized with trichloroacetic acid, and HPLC was performed on a column (25 cm x 4.6 mm) or Biophase ODS (5 pm) with a mobile phase comprising 0.1 M phosphate buffer (pH 3) and 0.34 mM Na octylsulfate at 1 mL/min. Detection was with a dual Hg-Au amalgam electrode versus a Ag-AgCl reference electrode. (z>)-penicillamine and homocysteine were determined at the downstream electrode at +0.15 V, and homocystine, penicillamine-homocysteine, and penicillamine disulfides were first reduced... 

Shaw et al. [64] described a (D)-penicillamine detection method in blood samples that had been treated with EDTA, deproteinized with trichloroacetic acid, and analyzed within 1 h. Penicillamine was detected at a vitreous-carbon electrode operated at +800 mV after HPLC separation. A linear calibration graph was obtained, and the method had a limit of detection equal to 5-20 ng. The method was useful in clinical and in pharmacokinetic studies. 

Parkhurst et al. [79] described a high performance liquid chromatographic method for the simultaneous determination of primaquine and its metabolites from plasma and urine samples, utilizing acetonitrile deproteinization, and direct injection onto a cyano column. Levels of 100 ng/mL per 20 pL injection could be quantitated. Preliminary pharmacokinetic analysis is reported for two human subjects after oral doses of 60 90 mg primaquine diphosphate. Two apparent plasma metabolites and two possible urinary metabolites are also reported. 

Dean et al. [93] used a high performance liquid chromatographic method for the simultaneous determination of primaquine and carboxyprimaquine in plasma with electrochemical detection. After the addition of the internal standard, plasma was deproteinized by the addition of acetonitrile. Nitrogen-dried supernatants, resuspended in mobile phase were analyzed on a C8 reversed-phase column. Limits of detection for primaquine and carboxyprimaquine were 2 and 5 ng/mL with quantitation limits of 5 and 20 ng/mL, respectively. The assay sensitivity and specificity are sufficient to permit quantitation of the drug in plasma for pharmacokinetics following low dose (30 mg, base) oral administration of primaquine, typically used in the treatment of malaria and P. carinii pneumonia. 

A competitive fluorescence-polarization immunoassay method was described for the monitoring of 12 drugs including valproic acid [18]. Samples (serum or plasma) were deproteinated. Fluorescence from the fluorescein-labeled analyte used as tracer was excited at 488 nm and polarization of light emitted at 531 nm was measured. The calibration was stable for 4 weeks and the coefficient of variation was below 10%. A single measurement took 8-10 min. 

The greatest area of applications of this type of ECL has been in the analysis of pharmaceutical compounds with amine functionality. The reader is directed toward the previously mentioned review articles and Table 1 for further details [12, 14-16], Many methods have also been successfully applied to real samples in the form of body fluids or pharmaceutical preparations, although sample pretreatment such as deproteinization, centrifugation, and neutralization followed by a chromatographic step to remove interfering species is often required. Limits of detection are typically in the range 10-9—10 12 M. Figure 4 shows examples of some classes of pharmaceutical compounds that have been determined by Ru(bpy)32+ ECL. 

More recent determinations of serum iron have been reported by Schmidt 57), who simply diluted with lanthanum chloride solution, and by Tavenier and Hellen-doorn58), who deproteinized samples in the latter study, iron in the protein precipitate is analyzed to correct the serum iron level. Uny etal. 59) determined serum iron, using ultrasonic nebulization of the sample to increase the sensitivity. Olson and Hamlin 6°) have determined serum iron and total iron-binding capacity. Proteins are precipitated and iron (III) is released by heating with trichloroacetic acid. 

There is a great deal of interest in the determination of lead, particularly micromethods applicable to the analysis blood lead in children. Consequently, reports continue to appear on the atomic absorption determination of lead in blood and urine. Ninety percent of blood lead is found in the erythrocytes and, therefore, whole blood is analyzed rather than serum or plasma. Berman etal. 134) have described a procedure for determining normal lead levels in which only 250 fd of blood are taken. The blood is deproteinized with 1 ml of 10 % trichloroacetic acid and then the lead is extracted with APDC into 1 ml of MIBK, at pH 3.5. 

Blood should be deproteinized by some technique which leaves no extra salt, acid, or alkali in the supernatant Some suitable techniques are with tungstic acid, with ethanol (BIO), or with zinc sulphate and barium hydroxide (S21). The supernatant is desalted in the same way as urine and, if necessary, concentrated before applying to the paper. Subsequent technique is as for urine. 

The first fractionation of urinary ampholytes in this way was carried out by Boulanger et al. (BIO) in 1952 with the use of ion-exchange resins. They had designed this procedure previously for the fractionation of ampholytes in blood serum (B8). According to this method, deproteinized urine was subjected to a double initial procedure aiming at the separation of low-molecular weight substances from macro-molecular ones. One of the methods consisted of the fractionation of urinary constituents by means of dialysis, the second was based on the selective precipitation of urinary ampholytes with cadmium hydroxide, which, as had previously been demonstrated, permits separation of the bulk of amino acids from polypeptides precipitated under these circumstances. Three fractions, i.e., the undialyzable part of urine, the dialyzed fraction, and the so-called cadmium precipitate were analyzed subsequently. 

In 1948 Dent (D2) described the so-called nephrosis peptide. He observed the presence of this peptide in the deproteinized urine of two patients with nephrosis. However, he could not find this peptide in the... 

In isolated nuclei from rat liver and kidney, Ni2 + was bound to chromatin, polynucleosomes and to deproteinized DNA [339]. Ni2+ directly interacted with stable binding sites on the DNA molecule in chromatin and was associated with histone and nonhistone nuclear proteins [339, 340]. 

Plasma Deproteinization with trichloroacetic acid bromination of supernatant and treatment with pyridine-p-phenylene diamine Spectrophotometry (thiocyanate- cyanide determination) 0.07 ppm 96 (thiocyanate) Pettigrew and Fell 1972... 

Carbamate derivatives (Table 1) of cellulose, chitin, amylose, amylopectin, and dextran were prepared using the isocyanates described in Part A of the Experimental Section. Amylose, amylopectin, dextran, and cellulose were obtained from Polysciences, Inc. and used without further purification. Chitin, obtained from Eastman Kodak, was decalcified and deproteinated by the method reported by Haye r prior to use. 

Vinyl alcohol copolymer gel is hydrophilic and has been developed for aqueous-phase size-exclusion liquid chromatography however, it is less polar than the polysaccharides. Its specificity permits the direct injection of a biological sample without deproteinization. For example, blood serum from a patient suffering from chronic nephritis has been injected directly as a measure of the degree of dialysis (Figure 3.17). Adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate in red blood cells have also been separated directly (Figure 3.18). Theophylline in blood serum has been... 

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