Two modifications of the carbazole reaction of hexuronic acids for the differentiation of polyuronides

Dische, Zacharias; Rothschild, Carl

doi:10.1016/0003-2697(67)90091-7

Cited by 27 publications

(10 citation statements)

References 4 publications

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“…The supernatant, obtained after the addition of Ba(OH)2 and ZnSO4, was lyophilized to dryness and the resulting residue, dissolved in sodium hydroxide, was applied to a Bio-Rad AG 1-X8 column in the acetate form and eluted with acetic acid (21). The fraction containing glucuronic acid was identified using carbazole (22) and evaporated to dryness. The glucuronic acid was further purified by applying it to a Aminex HPX 87H organic acid analysis column and eluting it under high pressure with water acidified to pH 2.5 with H2SO4.…”

Section: Methodsmentioning

confidence: 99%

Quantitation of the pathways of hepatic glycogen formation on ingesting a glucose load.

Magnusson

Chandramouli²,

Schumann³

et al. 1987

J. Clin. Invest.

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Diflunisal, 5-(2',4'-difluorophenyl)salicylic acid, excreted in urine as its glucuronide, was given to normal humans (n = 6) along with a glucose load specifically labeled with 14C. Glucuronide excreted by each subject was reduced to its glucoside and glucose from it degraded to yield the distribution of 14C in its six carbons. Randomization of the 14C from the specifically labeled glucose was taken as a measure of the extent to which glucose was deposited indirectly (i.e., glucose -) lactateglucose-6-P glycogen), rather than directly (i.e., glucose glucose-6-P glycogen). The maximum contribution to glycogen formation by the direct pathway was estimated to be 65±1%, on the assumption that glucuronide and glycogen are derived from the same hepatic pool of glucose-6-P in liver.Evidence that supports that assumption was obtained by comparing the randomization of 14C in the urinary glucuronide with that in glucose in blood from the hepatic vein of four of the subjects before and after they were given glucagon.

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Section: Methodsmentioning

confidence: 99%

Quantitation of the pathways of hepatic glycogen formation on ingesting a glucose load.

Magnusson

Chandramouli²,

Schumann³

et al. 1987

J. Clin. Invest.

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“…The column was washed with water and then increasing concentrations of acetic acid to 5 mol/l. The fractions in which the glucuronide eluted were identified using carbazole [35]. Most glucuronide eluted in the 5 mol/l fraction.…”

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confidence: 99%

Changes in hepatic glycogen cycling during a glucose load in healthy humans

et al. 2005

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Aims/hypothesis: Glycogen cycling, i.e. simultaneous glycogen synthesis and glycogenolysis, affects estimates of glucose fluxes using tracer techniques and may contribute to hyperglycaemia in diabetic conditions. This study presents a new method for quantifying hepatic glycogen cycling in the fed state. Glycogen is synthesised from glucose by the direct and indirect (gluconeogenic) pathways. Since glycogen is also synthesised from glycogen, i.e. glycogen→glucose 1-phosphate→glycogen, that synthesised through the direct and indirect pathways does not account for 100% of glycogen synthesis. The percentage contribution of glycogen cycling to glycogen synthesis then equals the difference between the sum of the percentage contributions of the direct and indirect pathways and 100. Materials and methods: The indirect and direct pathways were measured independently in nine healthy volunteers who had fasted overnight. They ingested 2 H 2 O (5 ml/kg body water) and were infused with [5-3 H] glucose and acetaminophen (paracetamol; 1 g) during hyperglycaemic clamps (7.8 mmol/l) lasting 8 h. The percentage contribution of the indirect pathway was calculated from the ratio of 2 H enrichments at carbon 5 to that at carbon 2, and the contribution of the direct pathway was determined from the 3 H-specific activity, relative to plasma glucose, of the urinary glucuronide excreted between 2 and 4, 4 and 6, and 6 and 8 h. Results: Glucose infusion rates increased (p<0.01) to ∼50 μmol kg −1 min −1 . Plasma insulin and the insulin : glucagon ratio rose ∼3.6-and ∼8.3-fold (p<0.001), respectively. From the difference between 100% and the sum of the direct (2-4 h, 54±6%; 4-6 h, 59±5%; 6-8 h, 63±4%) and indirect (32±3, 38±4, 36±3%) pathways, glycogen cycling was seen to be decreased (p<0.05) from 14±4% (2-4 h) to 4±3% (4-6 h) and 1±3% (6-8 h). Conclusions/interpretation: This method allows measurement of hepatic glycogen cycling in the fed state and demonstrates that glycogen cycling occurs most in the early hours after glucose loading subsequent to a fast.

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“…Besides the historic measurement of hexuronic acids (carbazole method: Dische 1947, Dische andRothschild 1967;Fig. 1 Tandem mass spectroscopy (MS/MS) triple quadrupole (Q) analyzer used in the multiple reaction monitoring (MRM) mode: Q1 selection of precursor ion, Q2 collision cell (fragmentation of precursor ion), Q3 selection of product ion(s) harmine method: Piraud et al 1993), the most widely used method in biochemical genetic laboratories for the quantification of total GAGs in urine is the 1,9-dimethylmethylene blue (DMB) spectrophotometric method involving the formation of a complex between DMB and sulfated GAGs (Whitley et al 1989a, b;deJongetal.1989deJongetal.…”

Section: Mucopolysaccharidosesmentioning

confidence: 99%

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

Piraud

Pettazzoni

Lavoie

et al. 2018

J of Inher Metab Disea

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Tandem mass spectrometry (MS/MS) is a highly sensitive and specific technique. Thanks to the development of triple quadrupole analyzers, it is becoming more widely used in laboratories working in the field of inborn errors of metabolism. We review here the state of the art of this technique applied to the diagnosis of lysosomal storage disorders (LSDs) and how MS/MS has changed the diagnostic rationale in recent years. This fine technology brings more sensitive, specific, and reliable methods than the previous biochemical ones for the analysis of urinary glycosaminoglycans, oligosaccharides, and sialic acid. In sphingolipidoses, the quantification of urinary sphingolipids (globotriaosylceramide, sulfatides) is possible. The measurement of new plasmatic biomarkers such as oxysterols, bile acids, and lysosphingolipids allows the screening of many sphingolipidoses and related disorders (Niemann-Pick type C), replacing tedious biochemical techniques. Applied to amniotic fluid, a more reliable prenatal diagnosis or screening of LSDs is now available for fetuses presenting with antenatal manifestations. Applied to enzyme measurements, it allows high throughput assays for the screening of large populations, even newborn screening. The advent of this new method can modify the diagnostic rationale behind LSDs.

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Two modifications of the carbazole reaction of hexuronic acids for the differentiation of polyuronides

Cited by 27 publications

References 4 publications

Quantitation of the pathways of hepatic glycogen formation on ingesting a glucose load.

Quantitation of the pathways of hepatic glycogen formation on ingesting a glucose load.

Changes in hepatic glycogen cycling during a glucose load in healthy humans

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

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