Perchloric acid, extraction

Fig. 12. Absorption spectra of perchloric acid extracts of whole blood from normal subject and a patient with pyrimidine 5 -nucleotides (P5N) deficiency. Absorption peak shift occurs in P5N deficiency, reflecting intracellular accumulation of pyrimidine nucleotides.

Fig. 4. 500 MHz DOSY spectrum of a D2O solution of a perchloric acid extract of gerbil brain. Assignments of selected signals are indicated as follows ac = acetate ala = alanine cho = choline cr = creatine ere = creatinine etn = ethanolamine GABA = 7-aminobutyric acid glu = glutamate GPC = glycerophosphocholine lac = lactate m-ino = myo-inositol NAA = N-acetylaspartate succ = succinate and tau = taurine. (Raw data...

AA and IAA simultaneously Yeast Metaphosphor ic acid/cold perchloric acid extraction Precolumn ODS-IO (40 X 2.6 mm Bio-Rad). Analytical Spherisorb ODS-2 (250 X 4.6 mm, 5 jum Rainin). 35°C. Isocratic 0.1 mM EDTA and 1.0 mM tetrabutyl-ammonium phosphate in 0.08 M acetate buffer, pH 4.2 + methanol (95 + 5, v/v). No flow rate reported. Electro- chemistry + 0.72 V vs. Ag/AgCl reference electrode, glassy carbon working electrode. External standardization. Linear range = 0-60 /ug/g yeast (dry weight). Reproducibility CV 2.3% for IAA, 1.2% for A A. Recoveries quantitative for both vitamers. 59... 

Enzyme hydrolysis, with papain, diastase, clarase, takadiastase, intestinal phosphatase, or combinations thereof is most commonly used to release pantothenate from food proteins (186). A cold perchloric acid extraction was used to release pantothenic acid from tissue samples (187). Food spoilage prior to analysis may lead to inflated pantothenic acid levels (19). 

Fig. (33). Agar diffusion plate of colon and normal tissue, n = normal tissue extract, c = CEA perchloric acid extract, t=KCl-HCl extract, At = immune serum against tumor, Ac = immune serum against CEA, Act=mixture of sera.

Fig. (34). Coupled electrophoresis - agar diffusion analysis of tumor extracts and immune serum A Perchloric acid extract (CEA, 1-4) normal extract (5) B.KCI-HCI extract (6-9).

O Neill and Sakamoto reported an enzymatic fluorimetric method for the determination of acetylcholine in biological extracts [41]. Nanomolar amounts of acetylcholine were determined in perchloric acid extracts of biological materials (brain tissues) by use of a system containing acetylcholineesterase, acetyl CoA synthetase, maleate dehydrogenase, and citrate synthase. The production of NADH2 was stoichiometrically related to the amount of acetylcholine in the system, and was followed fluorimetrically. Interfering fluorescent substances in the brain extracts were removed with acid-washed Florisil. 

Duan et al. reported the use of a rapid and simple method for the determination of acetylcholine and choline in mouse brain by high performance liquid chromatography, making use of an enzyme-loaded post column and an electrochemical detector [144]. Perchloric acid extracts of small brain tissue were injected onto the HPLC system with no prior clean-up procedure. Detection limits for both compounds were 1 pmol, and this method was successfully applied to the measurement of acetylcholine in discrete brain areas of the mouse. 

It should be pointed out (Kulaev et al., 1974a) that in Rh. rubrum, both in the dark and in the light, the accumulation of PolyP took place not only in salt-soluble but also in alkali-soluble and hot-perchloric-acid-extractible fractions. The total amounts of PolyP... 

The effect of starvation and the subsequent addition of phosphate-containing medium on phosphorus-containing compounds was studied by31P NMR spectroscopy of perchloric acid extracts and intact cells of Heterosigma akashiro (Watanabe et al., 1987, 1888, 1989). The PolyP content and chain length decreased under starvation and rapidly increased on P restoration in the medium (Table 8.8). 

Urine (phenols and cresols) Hydrolysis with perchloric acid extraction with diisopropyl ether GC/FID NR NR NIOSH 1974... 

The perchloric acid extraction procedure described by Kraus and Rein-both (K27) involves a perchloric acid extraction followed by neutralization with a potassium hydroxide solution. Riss et al. (R3) reported recoveries ranging from 68 to 87% for the adenosine-, cytidine-, guanosine-, and uridlne-5 -nucleotides, with a mean recovery of 78%. Other investigators have used the PCA extraction procedure in the analysis of nucleotides... 

This radioimmunoassay for CEA has been widely used clinically to quantitate the level of CEA in perchloric acid extracts of plasma. The sensitivity of this assay is approximately 0.5 ng of CEA per milliliter of plasma. Multiple analyses on the same sample have established standard deviations of 0.76 to 1.03 for plasma samples with CEA titers from 0 to 5.0 ng/ml, of 1.12 to 2.31 for plasma samples with titers from 5.1 to 10 ng/ml, and of 2.29 to 5.47 for samples with higher titers. a-Fetoprotein can be quantitated in serum samples of 25 fi using the Z-gel procedure however, the accuracy and reproducibility of the procedure have not been studied extensively. The sensitivity and reproducibility of the radioim-munoassy with Z-gel for insulin are comparable to those for other published procedures. ... 

Hydrolyze sample with perchloric acid extract phenol and cresol with... 

Adriaenssens et al. (A3) have described a simple screening method for the study of amino acids in tissues using frozen slices. Efron (ElO) has described methods for the extraction of free amino acids from urine, plasma, spinal fluid, sweat, and cellular material. Saifer (SI) has made a comparative study of seven published procedures for extracting free amino acids from brain tissue and concluded that perchloric acid extraction is the simplest and gives the most consistent results.With minor modifications this extraction method can be applied to amniotic fluid (S5), brain and other tissues (All, SI), plasma, urine, cerebrospinal fluid, etc. 

Fig. 6.22. The (a) 1-D and (b) 2-D-DOSY spectrum of a mixture containing D2O, glucose, ATP and SDS micelles [50]. (Adapted with permission from ref. [50]. Copyright 1993 American Chemical Society), (c) 1-D and (d) 2D-DOSY spectrum of the perchloric acid extract of a gerbil brain in D2O. Selected assignments are ac — acetate ala — alanine cho — choline cr — creatinine etn — ethanolamine GABA — y-aminobutyric acid glu — glutamine CPC — glycerophospho-choline lac — lactate m-ino — myo-inositol NAA — N-acetylaspartate succ — succinate and tau = taurine [51]. (Adapted with permission from ref. [51]. Copyright 1997 Elsevier.)...

Fig. 7.7 Analysis of Aventis inhibitor 7500 using the cellular lipolysis assay Isolated rat adipocytes were incubated with NBD-labeled dodecanoic acid (for incorporation into neutral lipid droplets) and then challenged with 1 pM isoproterenol (to stimulate lipolysis) prior to addition of increasing concentrations of inhibitor 7500. After incubation (1 h at 37°C), the adipocytes were separated from the medium by flotation through an oil layer. Portions of the medium were enzymatically analyzed for glycerol. Portions of the perchloric acid extracts from the adipocytes were analyzed in duplicate by thin-layer chromato-...

J.M. Carcellar, R Pascual, J.M. Howells, S.L. Mazucco, R. Griffiths, J.R. Pattern Recognition Analysis of H NMR Spectra from Perchloric Acid Extracts of Human Brain Tumour Biopsies, Magn. Res. Med. 39, 869-877 (1998). 

Vaux St. Cyr (VI) studied perchloric acid extracts of normal and of pathological sera. All of them contained seromucoid acid, and the latter sera contained in addition y-globulin, PsA-globulin, and albumin, but in variable amounts. 

Some studies have been undertaken in the area of cancer diagnosis using perchloric acid extracts of various types of human brain tumor tissue [47], and the spectra could be classified using neural network software giving -85% correct classification. Tissues can be studied by metabonomics through the MAS NMR technique, and published examples include prostate cancer [16,48], renal cell carcinoma [18], breast cancer [19, 49], and various brain tumors [50]. Other recent studies include an NMR-based urinary metabonomic study of multiple sclerosis in humans and non-human primates [51]. 

NMR spectra from perchloric acid extracts of human brain tumor biopsies. Magn. Reson. Med. 39 869-877. 

Samples were deproteinized with 0.2 M HC104- Before reaction with 0PA-thiol, a known amount of 3-3 -diaminodipropylamine was added to the tissue homogenates as an internal standard and perchloric acid extracts were adjusted to pH 9.5 by addition of borate buffer. For quantitation, polyamines were compared to the internal standard. The peak height ratios of polyamine to internal standard for a known concentration of polyamine was presented in Table 1. 

CLL Cells isolated from peripheral blood by density gradient centrifugation were cultured for 12 hr in the absence or presence of 1 iM CdA in nicotinamide-free RPMI medium, supplemented with L-glutamine 2 mM and 20% autologous plasma. Neutralized perchloric acid extracts were analyzed for NAD content by an enzyme cycling assay, and for ATP content by HPLC. 

Prior to analysis, perchloric acid extracts are neutralized with KOH (KCIO4 precipitates) and those made with trichloroacetic acid are extracted with ether to remove the acid with alcoholic extracts and those made with formic or acetic acid, the volatile precipitant is removed under vacuum. 

Many laboratories have succeeded in fractionating histones by selective extraction and differential precipitation. The selective extraction procedure is based on the difference in the amino acid composition of the various types of histones. For example, a histone-rich lysine can be obtained by extraction from whole thymus with 5% perchloric acid followed by precipitation with 10% trichloroacetic acid. If the histones found in the perchloric acid extract are passed through a carboxymethyl-cellulose column and eluted from the column with borate buffer at pH 9, then different components called 1, 2, and 3 can be recovered. 

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