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Clinical Chemistry 24: 1724-1729, 1978;
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Clinical Chemistry, Vol 24, 1724-1729, Copyright © 1978 by American Association for Clinical Chemistry

Enzymic fluorometric continuous-flow assays for blood glucose, lactate, pyruvate, alanine, glycerol, and 3-hydroxybutyrate

B Lloyd, J Burrin, P Smythe and KG Alberti

We describe enzymic fluorometric methods of automated analysis for glucose, lactate, pyruvate, 3-hydroxybutyrate, glycerol, and alanine in perchloric acid extracts of blood. Unmodified Technicon AutoAnalyzer II apparatus is used. The usual concentrations of all these metabolites can be measured in as little as 0.1 ml of blood from a fasting subject. Within-batch and between-batch coefficients of variation ranged from 0.4 to 4.4% for all metabolites except 3-hydroxybutyrate, for which CV's were higher for low concentrations. Analytical recovery of added metabolites ranged from 92 to 98%. Glucose, lactate, alanine, and 3- hydroxybutyrate are stable in perchloric acid extracts for at least 13 days at room temperature, and a year at -20 degrees C; pyruvate shows a 6--8% loss after 3 days and 52% by one year at -20 degrees C; glycerol concentrations were stable at -20 degrees C for at least 13 days. Blank fluorescence is found in perchloric acid extracts of blood, necessitating blank runs for pyruvate, 3-hydroxybutyrate, glycerol, and alanine. The systems are simple to use, relatively inexpensive to operate, and are recommended for any laboratory with high throughput of samples.


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P. Galassetti, A. R. Neill, D. Tate, A. C. Ertl, D. H. Wasserman, and S. N. Davis
Sexual Dimorphism in Counterregulatory Responses to Hypoglycemia after Antecedent Exercise
J. Clin. Endocrinol. Metab., August 1, 2001; 86(8): 3516 - 3524.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
D. S. Edgerton, S. Cardin, M. Emshwiller, D. Neal, V. Chandramouli, W. C. Schumann, B. R. Landau, L. Rossetti, and A. D. Cherrington
Small Increases in Insulin Inhibit Hepatic Glucose Production Solely Caused by an Effect on Glycogen Metabolism
Diabetes, August 1, 2001; 50(8): 1872 - 1882.
[Abstract] [Full Text] [PDF]

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 J. Appl. Physiol.Home page
P. Galassetti, S. Mann, D. Tate, R. A. Neill, D. H. Wasserman, and S. N. Davis
Effect of morning exercise on counterregulatory responses to subsequent, afternoon exercise
J Appl Physiol, July 1, 2001; 91(1): 91 - 99.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, S. Mann, D. Tate, R. A. Neill, F. Costa, D. H. Wasserman, and S. N. Davis
Effects of antecedent prolonged exercise on subsequent counterregulatory responses to hypoglycemia
Am J Physiol Endocrinol Metab, June 1, 2001; 280(6): E908 - E917.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
D. K. Levenhagen, J. D. Gresham, M. G. Carlson, D. J. Maron, M. J. Borel, and P. J. Flakoll
Postexercise nutrient intake timing in humans is critical to recovery of leg glucose and protein homeostasis
Am J Physiol Endocrinol Metab, June 1, 2001; 280(6): E982 - E993.
[Abstract] [Full Text] [PDF]

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 J. Clin. Endocrinol. Metab.Home page
S. N. Davis and D. Tate
Effects of Morning Hypoglycemia on Neuroendocrine and Metabolic Responses to Subsequent Afternoon Hypoglycemia in Normal Man
J. Clin. Endocrinol. Metab., May 1, 2001; 86(5): 2043 - 2050.
[Abstract] [Full Text]

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 J. Clin. Endocrinol. Metab.Home page
C. H. Gravholt, N. Møller, M. D. Jensen, J. S. Christiansen, and O. Schmitz
Physiological Levels of Glucagon Do Not Influence Lipolysis in Abdominal Adipose Tissue as Assessed by Microdialysis
J. Clin. Endocrinol. Metab., May 1, 2001; 86(5): 2085 - 2089.
[Abstract] [Full Text]

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 Am. J. Physiol. Endocrinol. Metab.Home page
C. M. Donmoyer, J. Ejiofor, D. B. Lacy, S.-S. Chen, and O. P. McGuinness
Fructose augments infection-impaired net hepatic glucose uptake during TPN administration
Am J Physiol Endocrinol Metab, May 1, 2001; 280(5): E703 - E711.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
S. Cardin, P. A. Jackson, D. S. Edgerton, D. W. Neal, C. S. Coffey, and A. D. Cherrington
Effect of Vagal Cooling on the Counterregulatory Response to Hypoglycemia Induced by a Low Dose of Insulin in the Conscious Dog
Diabetes, March 1, 2001; 50(3): 558 - 564.
[Abstract] [Full Text]

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 J. Clin. Endocrinol. Metab.Home page
M. C. Moore, A. D. Cherrington, S. L. Mann, and S. N. Davis
Acute Fructose Administration Decreases the Glycemic Response to an Oral Glucose Tolerance Test in Normal Adults
J. Clin. Endocrinol. Metab., December 1, 2000; 85(12): 4515 - 4519.
[Abstract] [Full Text]

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 Am. J. Physiol. Endocrinol. Metab.Home page
S. N. Davis, C. Shavers, and F. Costa
Differential gender responses to hypoglycemia are due to alterations in CNS drive and not glycemic thresholds
Am J Physiol Endocrinol Metab, November 1, 2000; 279(5): E1054 - E1063.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
A. E. Halseth, D. P. Bracy, and D. H. Wasserman
Limitations to basal and insulin-stimulated skeletal muscle glucose uptake in the high-fat-fed rat
Am J Physiol Endocrinol Metab, November 1, 2000; 279(5): E1064 - E1071.
[Abstract] [Full Text] [PDF]

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 JPEN J Parenter Enteral NutrHome page
S. Song Chen, C. Donmoyer, Y. Zhang, S. A. Hande, D. B. Lacy, and O. P. McGuinness
Impact of Enteral and Parenteral Nutrition on Hepatic and Muscle Glucose Metabolism
JPEN J Parenter Enteral Nutr, September 1, 2000; 24(5): 255 - 260.
[Abstract] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
M. G. Krishna, R. H. Coker, D. B. Lacy, B. A. Zinker, A. E. Halseth, and D. H. Wasserman
Glucagon response to exercise is critical for accelerated hepatic glutamine metabolism and nitrogen disposal
Am J Physiol Endocrinol Metab, September 1, 2000; 279(3): E638 - E645.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
C. M. Donmoyer, S.-S. Chen, S. A. Hande, D. B. Lacy, J. Ejiofor, and O. P. McGuinness
Hyperinsulinemia compensates for infection-induced impairment in net hepatic glucose uptake during TPN
Am J Physiol Endocrinol Metab, August 1, 2000; 279(2): E235 - E243.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
O. P. McGuinness, J. Ejiofor, D. B. Lacy, and N. Schrom
Hepatic glucose metabolism during intraduodenal glucose infusion: impact of infection
Am J Physiol Endocrinol Metab, July 1, 2000; 279(1): E108 - E115.
[Abstract] [Full Text] [PDF]

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 J. Clin. Endocrinol. Metab.Home page
S. N. Davis, C. Shavers, and F. Costa
Gender-Related Differences in Counterregulatory Responses to Antecedent Hypoglycemia in Normal Humans
J. Clin. Endocrinol. Metab., June 1, 2000; 85(6): 2148 - 2157.
[Abstract] [Full Text]

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 HypertensionHome page
F. Costa, J. Heusinkveld, R. Ballog, S. Davis, and I. Biaggioni
Estimation of Skeletal Muscle Interstitial Adenosine During Forearm Dynamic Exercise in Humans
Hypertension, May 1, 2000; 35(5): 1124 - 1128.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
R. H. Coker, D. B. Lacy, P. E. Williams, and D. H. Wasserman
Hepatic alpha - and beta -adrenergic receptors are not essential for the increase in Ra during exercise in diabetes
Am J Physiol Endocrinol Metab, March 1, 2000; 278(3): E444 - E451.
[Abstract] [Full Text] [PDF]

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 J. Clin. Endocrinol. Metab.Home page
S. Vigili de Kreutzenberg, C. Crepaldi, S. Marchetto, L. Calò, A. Tiengo, S. Del Prato, and A. Avogaro
Plasma Free Fatty Acids and Endothelium-Dependent Vasodilation: Effect of Chain-Length and Cyclooxygenase Inhibition
J. Clin. Endocrinol. Metab., February 1, 2000; 85(2): 793 - 798.
[Abstract] [Full Text]

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 J. Clin. Endocrinol. Metab.Home page
S. N. Davis, P. Galassetti, D. H. Wasserman, and D. Tate
Effects of Gender on Neuroendocrine and Metabolic Counterregulatory Responses to Exercise in Normal Man
J. Clin. Endocrinol. Metab., January 1, 2000; 85(1): 224 - 230.
[Abstract] [Full Text]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, Y. Koyama, R. H. Coker, D. B. Lacy, A. D. Cherrington, and D. H. Wasserman
Role of a negative arterial-portal venous glucose gradient in the postexercise state
Am J Physiol Endocrinol Metab, December 1, 1999; 277(6): E1038 - E1045.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
C. H. Gravholt, O. Schmitz, L. Simonsen, J. Bulow, J. S. Christiansen, and N. Moller
Effects of a physiological GH pulse on interstitial glycerol in abdominal and femoral adipose tissue
Am J Physiol Endocrinol Metab, November 1, 1999; 277(5): E848 - E854.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P.-S. Hsieh, M. C. Moore, B. Marshall, M. J. Pagliassotti, B. Shay, D. Szurkus, D. W. Neal, and A. D. Cherrington
The head arterial glucose level is not the reference site for generation of the portal signal in conscious dogs
Am J Physiol Endocrinol Metab, October 1, 1999; 277(4): E678 - E684.
[Abstract] [Full Text] [PDF]

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 Am. J. Clin. Nutr.Home page
B. E Wisse, L A. Campfield, E. B Marliss, J. A Morais, R. Tenenbaum, and R. Gougeon
Effect of prolonged moderate and severe energy restriction and refeeding on plasma leptin concentrations in obese women
Am. J. Clinical Nutrition, September 1, 1999; 70(3): 321 - 330.
[Abstract] [Full Text] [PDF]

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 J. Appl. Physiol.Home page
M. C. Lekas, S. J. Fisher, B. El-Bahrani, M. van Delangeryt, M. Vranic, and Z. Q. Shi
Glucose uptake during centrally induced stress is insulin independent and enhanced by adrenergic blockade
J Appl Physiol, August 1, 1999; 87(2): 722 - 731.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, C. A. Chu, D. W. Neal, G. W. Reed, D. H. Wasserman, and A. D. Cherrington
A negative arterial-portal venous glucose gradient increases net hepatic glucose uptake in euglycemic dogs
Am J Physiol Endocrinol Metab, July 1, 1999; 277(1): E126 - E134.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, R. H. Coker, D. B. Lacy, A. D. Cherrington, and D. H. Wasserman
Prior exercise increases net hepatic glucose uptake during a glucose load
Am J Physiol Endocrinol Metab, June 1, 1999; 276(6): E1022 - E1029.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P.-S. Hsieh, M. C. Moore, D. W. Neal, M. Emshwiller, and A. D. Cherrington
Rapid reversal of the effects of the portal signal under hyperinsulinemic conditions in the conscious dog
Am J Physiol Endocrinol Metab, May 1, 1999; 276(5): E930 - E937.
[Abstract] [Full Text] [PDF]

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 J. Appl. Physiol.Home page
D. A. Podolin, Y. Wei, and M. J. Pagliassotti
Effects of a high-fat diet and voluntary wheel running on gluconeogenesis and lipolysis in rats
J Appl Physiol, April 1, 1999; 86(4): 1374 - 1380.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
D. K. Sindelar, K. Igawa, C. A. Chu, J. H. Balcom, D. W. Neal, and A. D. Cherrington
Effect of a selective rise in hepatic artery insulin on hepatic glucose production in the conscious dog
Am J Physiol Endocrinol Metab, April 1, 1999; 276(4): E806 - E813.
[Abstract] [Full Text] [PDF]

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 J. Clin. Endocrinol. Metab.Home page
G. F. Lewis, A. Carpentier, D. Bilinski, A. Giacca, and M. Vranic
Counterregulatory Response to Hypoglycemia Differs According to the Insulin Delivery Route, But Does Not Affect Glucose Production in Normal Humans
J. Clin. Endocrinol. Metab., March 1, 1999; 84(3): 1037 - 1046.
[Abstract] [Full Text]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, K. S. Hamilton, F. K. Gibbons, D. P. Bracy, D. B. Lacy, A. D. Cherrington, and D. H. Wasserman
Effect of fast duration on disposition of an intraduodenal glucose load in the conscious dog
Am J Physiol Endocrinol Metab, March 1, 1999; 276(3): E543 - E552.
[Abstract] [Full Text] [PDF]

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 J. Appl. Physiol.Home page
A. E. Halseth, D. P. Bracy, and D. H. Wasserman
Limitations to exercise- and maximal insulin-stimulated muscle glucose uptake
J Appl Physiol, December 1, 1998; 85(6): 2305 - 2313.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
A. E. Halseth, N. Rheaume, A. B. Messina, E. K. Reed, M. G. Krishna, P. J. Flakoll, D. B. Lacy, and D. H. Wasserman
Regulation of hepatic glutamine metabolism during exercise in the dog
Am J Physiol Endocrinol Metab, October 1, 1998; 275(4): E655 - E664.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. Galassetti, M. Shiota, B. A. Zinker, D. H. Wasserman, and A. D. Cherrington
A negative arterial-portal venous glucose gradient decreases skeletal muscle glucose uptake
Am J Physiol Endocrinol Metab, July 1, 1998; 275(1): E101 - E111.
[Abstract] [Full Text] [PDF]

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 J. Appl. Physiol.Home page
P. Galassetti, F. K. Gibbons, K. S. Hamilton, D. B. Lacy, A. D. Cherrington, and D. H. Wasserman
Enhanced muscle glucose uptake facilitates nitrogen efflux from exercised muscle
J Appl Physiol, June 1, 1998; 84(6): 1952 - 1959.
[Abstract] [Full Text] [PDF]

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 Clin. Chem.Home page
R. Rej
They Use Enzymes for Everything!
Clin. Chem., June 1, 1998; 44(6): 1149 - 1153.
[Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
R. M. O'Doherty, A. E. Halseth, D. K. Granner, D. P. Bracy, and D. H. Wasserman
Analysis of insulin-stimulated skeletal muscle glucose uptake in conscious rat using isotopic glucose analogs
Am J Physiol Endocrinol Metab, February 1, 1998; 274(2): E287 - E296.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
P. A. Jackson, M. J. Pagliassotti, M. Shiota, D. W. Neal, S. Cardin, and A. D. Cherrington
Effects of vagal blockade on the counterregulatory response to insulin-induced hypoglycemia in the dog
Am J Physiol Endocrinol Metab, December 1, 1997; 273(6): E1178 - E1188.
[Abstract] [Full Text] [PDF]


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