Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer

Jean-Baptiste Roullet, B. Lacour, A. Ulmann, M. Bailly

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We describe a simple, rapid, and fully automated technique for measuring urinary pyrophosphates with a centrifugal analyzer (the ENI GEMSAEC). This technique depends on the enzymic magnesium-dependent reaction with UDPG pyrophosphorylase (UTP: α-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) and spectrophotometry of the NADPH formed in a combined system of phosphorylation and reduction. Many samples of urine can be analyzed quickly without pretreatment, with high sensitivity (1.3 mA/μmol of substrate) and good reproducibility. The mean within-run coefficient of variation for a 50 μmol/L pyrophosphate solution was 1.4%. We determined the optimum enzyme and magnesium concentrations necessary for use in a 4-min reaction. Because there is no inhibitory effect of chloride and phosphate ions, pyrophosphate can be measured directly in urine, without prior extraction. With this technique, the mean value (and SD) for urinary pyrophosphate excretion by 30 healthy subjects was 39.3 (SD 17.2) μmol/24 h.

Original languageEnglish (US)
Pages (from-to)134-137
Number of pages4
JournalClinical Chemistry
Volume28
Issue number1
StatePublished - 1982
Externally publishedYes

Fingerprint

Magnesium
UTP-Glucose-1-Phosphate Uridylyltransferase
Urine
Uridine Triphosphate
Phosphorylation
Diphosphates
Spectrophotometry
Transferases
NADP
Chlorides
Healthy Volunteers
Phosphates
Ions
Glucose
Substrates
Enzymes
diphosphoric acid
glucose-1-phosphate

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Roullet, J-B., Lacour, B., Ulmann, A., & Bailly, M. (1982). Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer. Clinical Chemistry, 28(1), 134-137.

Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer. / Roullet, Jean-Baptiste; Lacour, B.; Ulmann, A.; Bailly, M.

In: Clinical Chemistry, Vol. 28, No. 1, 1982, p. 134-137.

Research output: Contribution to journalArticle

Roullet, J-B, Lacour, B, Ulmann, A & Bailly, M 1982, 'Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer', Clinical Chemistry, vol. 28, no. 1, pp. 134-137.
Roullet J-B, Lacour B, Ulmann A, Bailly M. Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer. Clinical Chemistry. 1982;28(1):134-137.
Roullet, Jean-Baptiste ; Lacour, B. ; Ulmann, A. ; Bailly, M. / Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer. In: Clinical Chemistry. 1982 ; Vol. 28, No. 1. pp. 134-137.
@article{dabec75da89540beb0506b6bf99d98b2,
title = "Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer",
abstract = "We describe a simple, rapid, and fully automated technique for measuring urinary pyrophosphates with a centrifugal analyzer (the ENI GEMSAEC). This technique depends on the enzymic magnesium-dependent reaction with UDPG pyrophosphorylase (UTP: α-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) and spectrophotometry of the NADPH formed in a combined system of phosphorylation and reduction. Many samples of urine can be analyzed quickly without pretreatment, with high sensitivity (1.3 mA/μmol of substrate) and good reproducibility. The mean within-run coefficient of variation for a 50 μmol/L pyrophosphate solution was 1.4{\%}. We determined the optimum enzyme and magnesium concentrations necessary for use in a 4-min reaction. Because there is no inhibitory effect of chloride and phosphate ions, pyrophosphate can be measured directly in urine, without prior extraction. With this technique, the mean value (and SD) for urinary pyrophosphate excretion by 30 healthy subjects was 39.3 (SD 17.2) μmol/24 h.",
author = "Jean-Baptiste Roullet and B. Lacour and A. Ulmann and M. Bailly",
year = "1982",
language = "English (US)",
volume = "28",
pages = "134--137",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "1",

}

TY - JOUR

T1 - Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer

AU - Roullet, Jean-Baptiste

AU - Lacour, B.

AU - Ulmann, A.

AU - Bailly, M.

PY - 1982

Y1 - 1982

N2 - We describe a simple, rapid, and fully automated technique for measuring urinary pyrophosphates with a centrifugal analyzer (the ENI GEMSAEC). This technique depends on the enzymic magnesium-dependent reaction with UDPG pyrophosphorylase (UTP: α-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) and spectrophotometry of the NADPH formed in a combined system of phosphorylation and reduction. Many samples of urine can be analyzed quickly without pretreatment, with high sensitivity (1.3 mA/μmol of substrate) and good reproducibility. The mean within-run coefficient of variation for a 50 μmol/L pyrophosphate solution was 1.4%. We determined the optimum enzyme and magnesium concentrations necessary for use in a 4-min reaction. Because there is no inhibitory effect of chloride and phosphate ions, pyrophosphate can be measured directly in urine, without prior extraction. With this technique, the mean value (and SD) for urinary pyrophosphate excretion by 30 healthy subjects was 39.3 (SD 17.2) μmol/24 h.

AB - We describe a simple, rapid, and fully automated technique for measuring urinary pyrophosphates with a centrifugal analyzer (the ENI GEMSAEC). This technique depends on the enzymic magnesium-dependent reaction with UDPG pyrophosphorylase (UTP: α-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) and spectrophotometry of the NADPH formed in a combined system of phosphorylation and reduction. Many samples of urine can be analyzed quickly without pretreatment, with high sensitivity (1.3 mA/μmol of substrate) and good reproducibility. The mean within-run coefficient of variation for a 50 μmol/L pyrophosphate solution was 1.4%. We determined the optimum enzyme and magnesium concentrations necessary for use in a 4-min reaction. Because there is no inhibitory effect of chloride and phosphate ions, pyrophosphate can be measured directly in urine, without prior extraction. With this technique, the mean value (and SD) for urinary pyrophosphate excretion by 30 healthy subjects was 39.3 (SD 17.2) μmol/24 h.

UR - http://www.scopus.com/inward/record.url?scp=0020061433&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020061433&partnerID=8YFLogxK

M3 - Article

C2 - 6276045

AN - SCOPUS:0020061433

VL - 28

SP - 134

EP - 137

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 1

ER -