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A L Yergey

Publications and source records attributed to A L Yergey.

85 records · Page 5Linked to original sources
PubMed1986

Separation and quantification of choline and acetylcholine by thermospray liquid chromatography/mass spectrometry.

The development of sensitive analytical techniques for use in the identification and qualification of molecules of biological origin remains a constant challenge. Often the compounds of interest are present in a complex mixture and require extensive sample preparation prior to analysis. Thermospray liquid chromatography/mass spectrometry (LC/MS) has been shown to be a method which allows for both separation of target molecules from complex mixtures and sensitive specific detection using a mass spectrometer. In this work thermospray LC/MS is used for the direct detection of intact, underivatized choline and acetylcholine from mouse brain homogenate. The results of our analysis corroborate previous analyses using a variety of indirect measurement techniques and thereby show that this simple direct analysis has wide potential applicability.

Acetylcholine↗
PubMed1985

Valproylcarnitine: a novel drug metabolite identified by fast atom bombardment and thermospray liquid chromatography-mass spectrometry.

Urine samples from three children at different stages of chronic valproate therapy were partially purified using a cation exchange column. A signal consistent with either valproylcarnitine or octanoylcarnitine was observed in one of these extracts by direct fast atom bombardment-mass spectrometry analysis. These isomeric acylcarnitines were synthesized, separated and characterized by thermospray high performance liquid chromatography-mass spectrometry. This new technique was then employed to positively identify intact valproyl-carnitine in the patients' urine samples. The implications of this finding with regard to a mechanism to account for carnitine deficiency in patients receiving valproate are discussed.

Adolescent↗
PubMed1985

Dynamics of calcium metabolism in infancy and childhood. I. Methodology and quantification in the infant.

The dynamics of calcium metabolism in the human infant were determined using stable isotopes of calcium as tracers. Two isotopes, one given intravenously and the other orally were used. Tracer time-dependent dilutions were measured with high accuracy and precision using thermal ionization mass spectrometry. Similar studies were performed in infant rhesus monkeys using radio tracers. Analysis of the isotope dilutions using a compartmental model method showed quite similar results for the infants of both species. These similarities support the validity both of using stable isotopes in human studies, and the use of the rhesus monkey as a model for human calcium dynamics. It was found that the classical steady state compartmental model was inadequate for use in the rapidly growing infant, and the model was modified to account for the expansion of the body fluid calcium compartment during the time of the study. Comparison with adult human calcium metabolic parameters showed that urinary calcium flows are much higher in the adult than in the newborn, relative to other calcium metabolic parameters. Other parameters of infant calcium metabolism are all consistent with the growth and more rapid metabolism usually seen in the young.

Absorption↗
PubMed1984

The biosynthesis of hypusine (N epsilon-(4-amino-2-hydroxybutyl)lysine). Alignment of the butylamine segment and source of the secondary amino nitrogen.

The unusual amino acid hypusine is produced in a single protein of mammalian cells by a novel posttranslational event in which a lysine residue is conjugated with the four-carbon moiety from the polyamine spermidine to form an intermediate deoxyhypusine, and in which this intermediate is subsequently hydroxylated. Specifically isotopically labeled precursors of hypusine were used to identify the biosynthetic origin of some of the atoms of hypusine and thus to provide further insight into the mechanism of this in vivo chemical modification reaction. Radiolabel from [1,4-3H] putrescine, [1,8-3H]spermidine, and [5-3H]spermidine entered hypusine during growth of Chinese hamster ovary cells. The occurrence of this label at positions 1 and 4, at position 4, and at position 1, respectively, in the 4-amino-2-hydroxybutyl portion of hypusine revealed an alignment of atoms identical to that in the butylamine segment of spermidine. Growth of cells with [epsilon-15N]lysine as the source of lysine yielded hypusine enriched in 15N, whereas only isotope-free hypusine during growth by [4-15N]spermidine. These was found in cells whose spermidine was replaced during growth by [4-15N]spermidine. These findings are in accordance with a proposal that the first phase of hypusine biosynthesis, the production of intermediate deoxyhypusine, occurs through transfer of the butylamine moiety from spermidine to the epsilon-amino nitrogen of protein-bound lysine. The technique of thermospray high-performance liquid chromatography/mass spectrometry provided positive identification of 15N in hypusine through final separation and on-column direct analysis of this amino acid. Methods of preparation are given for spermidine of high specific radioactivity, labeled specifically at position 5 with 3H, and for spermidine with 15N at the 4-position.

Animals↗
PubMed1984

Thermospray liquid chromatography/mass spectrometry for the analysis of L-carnitine and its short-chain acyl derivatives.

A method utilizing thermospray high-performance liquid chromatography/mass spectrometry for the separation and direct analysis of carnitine, acetylcarnitine, and propionylcarnitine is described. On-column analysis of mixtures of the acylcarnitines with their corresponding stable, isotope-labeled analogs at nanomolar concentrations has indicated that isotope dilution assays can be applied towards the analysis of carnitine and short-chain acylcarnitines present in biological samples.

Acetylcarnitine↗
PubMed1981

Quantitative analysis of disaturated lecithins in human plasma by ammonia chemical ionization mass spectrometry.

Three physiologically important disaturated lecithins have been analyzed quantitatively as the acetate derivatives using a solids inlet probe and ammonia chemical ionization. Concentration independent response factors have been determined over a tenfold range that brackets human plasma levels. The results obtained serve as an independent corroboration of gas chromatographic analyses.

Chromatography, Gas↗
PubMed1978

Monitoring normal and malignant human white blood cells by the use of linear programmed thermal degradation mass spectrometry.

A new methodology has been applied to the analysis of human white blood cells obtained both from donors with normal white cells and from donors having one of several cytologically identified leukemias. The analytical process makes use of evolution patterns of molecular fragments generated during the well controlled degradation of the cells. The process has been shown to be effective both in grouping together malignant cells that originate from the same diseases and in distinguishing between malignant cells originating from different diseases. The reproducibility of the method has been demonstrated.

Computers↗
PubMedDate not supplied

Furosemide 1-O-acyl glucuronide. In vitro biosynthesis and pH-dependent isomerization to beta-glucuronidase-resistant forms.

A biosynthetic acyl-type glucuronic acid conjugate of furosemide was isolated from in vitro incubation of pregnenolone-16 alpha-carbonitrile-induced rat liver microsomes containing UDP-glucuronyltransferase activity, furosemide, and UDP-glucuronic acid. Furosemide 1-O-acyl glucuronide (FG) was specifically hydrolyzed by beta-glucuronidase (BG) and was also labile to alkaline hydrolysis. FG concentration decreased at an apparent first order rate when incubated at 37 degrees C in buffer solution of pH values greater than 6.0 with only moderate hydrolysis of the conjugate at pH values less than 8.5. Formation of rearrangement forms of FG that were resistant to BG but labile to alkaline hydrolysis accounted for most of the disappearance of FG at this pH range. Radiochemical labeling of the conjugate with either 14C-furosemide or 14C-UDP-glucuronic acid was detected in the BG-resistant isomerization products of FG as they were separated by HPLC. The structure of FG and its isomerization products was further verified by negative ion thermospray liquid chromatography/mass spectrometry. The abundant (M - 1)-ion at mass 505, the aglycone fragment at m/z 329, and the characteristic sugar fragment ion of mass 175 were found in the spectra of FG and three additional isomers. An ion at m/z 221 was noted only in the case of the parent conjugate and thus may prove to be a characteristic ion for 1-O-acyl-linked glucuronides under negative ion thermospray. In vivo as well as in vitro rearrangement of FG to BG-resistant forms might affect the results of furosemide disposition studies which use BG hydrolysis to determine FG formation.

Animals↗