Abstract
It was studied that making conditions of a micro-volume fluorescence capillary biosensor for determining pyruvate (PA) and lactate (LA). The biosensor made under the optimized conditions could be used for sequential quantifications of LA in the range 0.10–1.2 mM and PA in 4–120 μM, and its recovery for PA and LA was in a satisfactory range 97–106% for human serum samples, with detection limits of 0.023 mM for LA (RSD < 1.89%, n = 11) and 0.87 μM for PA (RSD < 1.70%, n = 11). The new assay possessed these advantages that the LDH immobilizing on capillary realized the reuse of expensive enzyme in fluorospectrophotometry, and the consumption of serum samples or chemical reagents decreased to 9 μL in per assay, and the analytes no needed to preseparation, and it also are accurate and reliable. Consequently, the fluorescence capillary biosensor should have a good prospect in assaying PA and LA or LA/PA ratios for clinical medicines or biology field. The optimization conditions and parameters obtained in this study have also a certain guiding significance for the development of biochip based on glass substrate.
Similar content being viewed by others
References
Burtis CA, Ashwood ER, Bruns DE (2011) Tietz Textbook of clinical chemistry and molecular diagnostics. 5nd Edit, Saunders, Elsevier Medicine
Okada H, Araga S, Takshima T, Nakashima K (1998) Plasma lactic acid and pyruvic acid levels in migraine and tension-type headache. Headache 38(1):39–42
Tsao YS, Cardoso AG, Condon RGG, Voloch M, Lio P, Lagos JC, Kearns BG, Liu Z (2005) Monitoring Chinese hamster ovary cell culture by the analysis of glucose and lactate metabolism. J Biotechnol 118:316–327
Hofstede HJM, Willems HL, Koopmansa P (2004) Serum L-lactate and pyruvate in HIV-infected patients with and without presumed NRTI-related adverse events compared to healthy volunteers. J Clin Virology 29:44–50
Mackay N, Robinson BH (2007) Measurement of the ratio of lactate to pyruvate in skin fibroblast cultures. Methods Cell Biol 80:173–178
Low JA, Pancham SR, Worthington D, Boston RW (1974) Acid-base, lactate, and pyruvate characteristics of the normal obstetric patient and fetus during the intrapartum period. Am J Obstet Gynecol 120:862–867
Kala G, Hertz L (2005) Ammonia effects on pyruvate/lactate production in astrocytes--interaction with glutamate. Neurochem Int 47:4–12
Hatherill M, Salie S, Waggie Z, Lawrenson J, Hewitson J, Reynolds L, Argent A (2007) The lactate: pyruvate ratio following open cardiac surgery in children. Intensive Care Med 33:822–829
Persson L, Hillered L (1992) Chemical monitoring of neurosurgical intensive care patients using intracerebral microdialysis. J Neurosurg 76:72–80
Wu YS, Tsai TH, Wu TF, Cheng FC (2001) Determination of pyruvate and lactate in primary liver cell culture medium during hypoxia by on-line microdialysis–liquid chromatography. J Chromato A 913:341–347
Chuang CK, Wang TJ, Yeung CY, Lin DS, Lin HY, Liu HL, Ho HT, Hsieh WS, Lin SP (2009) A method for lactate and pyruvate determination in filter-paper dried blood spots. J Chromatog A 1216:8947–8952
Gu YH, Kodama H, Ogawa E, Izumi Y (2014) Lactate and pyruvate levels in blood and cerebrospinal fluid in patients with Menkes disease. J Pediatr 164:890–894
Hallstrom A, Carlsson A, Hillered L, Uncerstedt U (1989) Simultaneous determination of lactate, pyruvate, and ascorbate in microdialysis samples from rat brain, blood, fat, and muscle using high-performance liquid chromatography. J Pharmacol Methods 22(2):113–124
Cheng FC, Yang LL, Yang DY, Tsai TH, Lee CW, Chen SH (2000) Monitoring of extracellular pyruvate, lactate, and ascorbic acid during cerebral ischemia: a microdialysis study in awake gerbils. J Chromatog A 870:389–394
Minniti G, Cerone R, De Toni E (2001) Determination of lactic acid, pyruvic acid, and ketone bodies in serum and cerebrospinal fluid by HPLC. Am Clin Lab 20:21–23
Paik MJ, Cho EY, Kim H, Kim KR, Choi S, Ahn YH, Lee G (2008) Simultaneous clinical monitoring of lactic acid, pyruvic acid and ketone bodies in plasma as methoxime/tert-butyldimethylsilyl derivatives by gas chromatography-mass spectrometry in selected ion monitoring mode. Biomed Chromatogr 22:450–453
Yao T, Yano T (2004) On-line microdialysis assay of L-lactate and pyruvate in vitro and in vivo by a flow-injection system with a dual enzyme electrode. Talanta 63:771–775
Baena B, Garcia-Martinez D, Barbas C (2004) Evaluation of diabetes-related short-chain organic acids in rat plasma by capillary electrophoresis. J Chromatogr A 1051:199–205
Xue QF, Yeung ES (1994) Indirect fluorescence determination of lactate and pyruvate in single erythrocytes by capillary electrophoresis. J Chromatogr A 661:287–295
Olsen C (1971) An enzymic fluorometric micromethod for the determination of acetoacetate, β-hydroxybutyrate, pyruvate, and lactate. Clin Chim Acta 33:293–300
Neville JF, Gelder RL (1971) Modified enzymatic methods for the determination of L-(+)-lactic and pyruvic acids in blood. Am J Clin Pathol 55:152–158
Maughan RJ (1982) A simple, rapid method for the determination of glucose, lactate, pyruvate, alanine, 3-hydroxybutyrate and acetoacetate on a single 20-μl blood sample. Clin Chim Acta 122:231–240
Artuch R, Vilaseca MA, Farre C, Ramon F (1995) Determination of lactate, pyruvate, beta-hydroxybutyrate and acetoacetate with a centrifugal analyzer. Eur J Clin Chem Clin Biochem 33:529–533
Hansen JL, Freier EF (1978) Direct assays of lactate, pyruvate, β-hydroxybutyrate, and acetoacetate with a centrifugal analyzer. Clin Chem 24:475–479
Kopperschlager G, Kirchberger J (1996) Methods for the separation of lactate dehydrogenases and clinical significance of the enzyme. J Chromatogr B 684:25–49
Li YS, Gao XF (2007) A new method for chemical analysis: fluorescence capillary analysis. Spectrosc Spect Anal 27:1565–1569
Li YS, Ju X, Gao XF, Zhao YY, Wu YF (2008) Immobilization enzyme fluorescence capillary analysis for determination of lactic acid. Anal Chim Acta 610:249–256
Li QJ, Gao XF, Du YD, Chen TM, Li YS (2011) Determination of blood glucose by fluorescence capillary analysis based on glucose/ glucose oxidase/H2O2/L-tyrosine/ horse radish peroxidase reaction system. Chinese J Anal Chem 39:1181–1185
Gao XF, Li YS, Jiang YX (2006) Research on fluorescence capillary analysis of DNA marked by using Cy-5 reagent. Chinese J Anal Chem 34:S220–S222
Wang YJ, Li YS, Yang QY, Huang Y, Tang J, Gao XF (2009) Research on DNA fluorescence capillary biosensor marked by goldview. Spectrosc Spect Anal 29:165–168
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Li, YS., Li, QJ., Yang, W. et al. Research on a New Micro-Volume Fluorescence Capillary Biosensor Assay for Sequentially Quantifying Pyruvate and Lactate. J Fluoresc 27, 883–894 (2017). https://doi.org/10.1007/s10895-017-2024-3
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1007/s10895-017-2024-3