Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells

Steven L. Mitchell; Brian D. Ross; Thomas Krick; Michael Garwood

doi:10.1016/S0006-291X(89)80072-5

Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells

Steven L. Mitchell, Brian D. Ross, Thomas Krick, Michael Garwood

Radiology

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

A GC/MS method is described for monitoring the relative amount of glucose degraded to lactate via the hexose monophosphate shunt (HMPS) in neoplastic cells. C6 glioma cells were incubated in medium supplemented with [1-¹³C]glucose and medium containing [1-¹³C]glucose with 0.001mM phenazine methosulfate (PMS). The ratio of the [¹³C]lactate to [¹²C]lactate determined from the measurement of the m/z 219/220 and 117/118 ions of the trimethyl silyl derivative, was used to calculate HMPS activity. PMS increased HMPS activity in C6 glioma cells by 3.2 and 4.8 fold at 2 and 12 hours of incubation respectively. GC/MS results were compared with ¹H NMR measurements of the [3-¹³C]lactate/[3-¹²C]lactate ratio. The GC/MS method was found to require less sample size and yielded better sensitivity than the NMR method.

Original language	English (US)
Pages (from-to)	474-479
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	158
Issue number	2
DOIs	https://doi.org/10.1016/S0006-291X(89)80072-5
State	Published - Jan 31 1989

Bibliographical note

Funding Information:
This work was supported by funds from the Department of Radiology, University of Minnesota. BDR is a Fellow of the Association For Brain Tumor Research.

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title = "Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells",

abstract = "A GC/MS method is described for monitoring the relative amount of glucose degraded to lactate via the hexose monophosphate shunt (HMPS) in neoplastic cells. C6 glioma cells were incubated in medium supplemented with [1-13C]glucose and medium containing [1-13C]glucose with 0.001mM phenazine methosulfate (PMS). The ratio of the [13C]lactate to [12C]lactate determined from the measurement of the m/z 219/220 and 117/118 ions of the trimethyl silyl derivative, was used to calculate HMPS activity. PMS increased HMPS activity in C6 glioma cells by 3.2 and 4.8 fold at 2 and 12 hours of incubation respectively. GC/MS results were compared with 1H NMR measurements of the [3-13C]lactate/[3-12C]lactate ratio. The GC/MS method was found to require less sample size and yielded better sensitivity than the NMR method.",

author = "Mitchell, {Steven L.} and Ross, {Brian D.} and Thomas Krick and Michael Garwood",

note = "Funding Information: This work was supported by funds from the Department of Radiology, University of Minnesota. BDR is a Fellow of the Association For Brain Tumor Research.",

year = "1989",

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doi = "10.1016/S0006-291X(89)80072-5",

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AU - Mitchell, Steven L.

AU - Ross, Brian D.

AU - Krick, Thomas

AU - Garwood, Michael

N1 - Funding Information: This work was supported by funds from the Department of Radiology, University of Minnesota. BDR is a Fellow of the Association For Brain Tumor Research.

PY - 1989/1/31

Y1 - 1989/1/31

N2 - A GC/MS method is described for monitoring the relative amount of glucose degraded to lactate via the hexose monophosphate shunt (HMPS) in neoplastic cells. C6 glioma cells were incubated in medium supplemented with [1-13C]glucose and medium containing [1-13C]glucose with 0.001mM phenazine methosulfate (PMS). The ratio of the [13C]lactate to [12C]lactate determined from the measurement of the m/z 219/220 and 117/118 ions of the trimethyl silyl derivative, was used to calculate HMPS activity. PMS increased HMPS activity in C6 glioma cells by 3.2 and 4.8 fold at 2 and 12 hours of incubation respectively. GC/MS results were compared with 1H NMR measurements of the [3-13C]lactate/[3-12C]lactate ratio. The GC/MS method was found to require less sample size and yielded better sensitivity than the NMR method.

AB - A GC/MS method is described for monitoring the relative amount of glucose degraded to lactate via the hexose monophosphate shunt (HMPS) in neoplastic cells. C6 glioma cells were incubated in medium supplemented with [1-13C]glucose and medium containing [1-13C]glucose with 0.001mM phenazine methosulfate (PMS). The ratio of the [13C]lactate to [12C]lactate determined from the measurement of the m/z 219/220 and 117/118 ions of the trimethyl silyl derivative, was used to calculate HMPS activity. PMS increased HMPS activity in C6 glioma cells by 3.2 and 4.8 fold at 2 and 12 hours of incubation respectively. GC/MS results were compared with 1H NMR measurements of the [3-13C]lactate/[3-12C]lactate ratio. The GC/MS method was found to require less sample size and yielded better sensitivity than the NMR method.

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Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells

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