Determination of pyruvic acid by using enzymic fluorescence capillary analysis

Zhao, Yuanyuan; Gao, Xiu-Feng; Li, Yongsheng; Xiang, Jun; Zhang, Jia; Zheng, Jinchuan

doi:10.1016/j.talanta.2008.02.031

Cited by 22 publications

(13 citation statements)

References 21 publications

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“…Moreover, to further evaluate the validity of the proposed method, a comparative experiment was also conducted by an enzymatic fluorometric method (Table 1). [20,21] The results from the two methods are in good agreement with each other, and no significant difference between them is found at 90 % confidence level when using the paired t test. In addition, the relative standard derivation (about 1 %) of the present method is lower than that (about 5 %) of the enzymatic fluorometric assay, indicating that the RBPH-based method has better precision.…”

Section: Linearitysupporting

confidence: 63%

“…For enzymatic fluorimetric analysis, the solutions of NADH (2 mm) and LDH (5 KUL À1 ; one unit of the enzyme activity was defined as the amount of the enzyme that reduced 1.0 mmol of pyruvate to l-lactate per min at pH 7.5 at 37 8C were prepared daily by dissolving appropriate amounts of the compounds in 0.2 m phosphate buffer (pH 7.5) and stored at 4 8C. [20,21] 1 H and 13 C NMR spectra were measured on a Bruker DMX-300 spectrometer at 300 and 75 MHz, respectively, in D 2 O. Electrospray ionization (ESI) mass spectra were recorded with a Shimadzu LCMS-2010. High-resolution SIMS analysis was performed by using a Bruker-Daltonics APEX II FT-ICR instrument.…”

Section: Methodsmentioning

confidence: 99%

“…[20] Amperometric biosensors are rapid and sensitive, but suffer from interference. [23] The mass spectrometric and enzymatic methods show good selectivity and sensitivity; both of them would be more promising if their relative high cost can be reduced.…”

Section: Linearitymentioning

confidence: 99%

“…[23] The mass spectrometric and enzymatic methods show good selectivity and sensitivity; both of them would be more promising if their relative high cost can be reduced. [20] Because of the improved spectroscopic properties and labeling conditions of the reagent RBPH, our proposed method may offer a new choice for the simple, selective, and sensitive determination of pyruvic acid. Moreover, to further evaluate the validity of the proposed method, a comparative experiment was also conducted by an enzymatic fluorometric method (Table 1).…”

Section: Linearitymentioning

confidence: 99%

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Rhodamine B Piperazinoacetohydrazine: A Water‐Soluble Spectroscopic Reagent for Pyruvic Acid Labeling

Jia

Wang

Chen

et al. 2010

Chemistry A European J

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A new water-soluble reagent, rhodamine B piperazinoacetohydrazine (RBPH), with improved spectroscopic and reaction properties, has been developed and characterized for pyruvic acid labeling. The reagent RBPH is designed and synthesized by using rhodamine B as a spectroscopic unit, and hydrazine as a carbonyl-specific labeling unit; the two units are connected by a well-chosen linker of piperazine, which prohibits the formation of the nonfluorescent spirocyclic structure of rhodamine B, thereby keeping the spectroscopic response of the reagent in a stable state. Such a design enables RBPH not only to maintain its excellent spectroscopic properties over a wide pH range, but also to exist as a stable cation with high water solubility. Moreover, the hydrazino group of RBPH is expected to react selectively with carbonyl compounds under mild conditions through the rapid formation of hydrazones. These important features make RBPH of great potential use in the labeling of aldehydes or ketones in various biosystems, and such an application of RBPH as a precolumn derivatizing reagent has been successfully demonstrated on the analysis of pyruvic acid in human serum by high-performance liquid chromatography with common UV/Vis detection.

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Section: Linearitysupporting

confidence: 63%

Section: Methodsmentioning

confidence: 99%

Section: Linearitymentioning

confidence: 99%

Section: Linearitymentioning

confidence: 99%

See 2 more Smart Citations

Rhodamine B Piperazinoacetohydrazine: A Water‐Soluble Spectroscopic Reagent for Pyruvic Acid Labeling

Jia

Wang

Chen

et al. 2010

Chemistry A European J

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“…This method is very suitable for precious and less samples (or reagents) to assay. Based on the FCA, some new determination method has been published for micro-volume alcohol, 23 DNA, 24 lactic acid, 25 pyruvate, 26 trace-level sulfated bile acid in urine 27 and blood glucose, 28 however, the feasibility of using the FCA method to quantify enzyme activity in a micro-volume sample is still not confirmed. Therefore, we carried out this research in which the LDH activity was determined by the FCA method based on fluorescent NADH (λex/λem 350 nm/460 nm) to non-fluorescent NAD + .…”

Section: Introductionmentioning

confidence: 99%

A Novel Analysis Method for Lactate Dehydrogenase Activity in Serum Samples Based on Fluorescence Capillary Analysis

Gao

2015

ANAL. SCI.

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Based on fluorescence capillary analysis technology, a method for quantitating lactate dehydrogenase (LDH) activity in a micro-volume sample was developed. Sample and reagent consumptions were merely 2 and 16 μL per time, respectively. The optimized test conditions were as follows. The reaction reagent consisted of 0.10 M phosphate buffer (pH 6.5), 0.30 mM NADH and 1.20 mM pyruvate. NADH standard was prepared with a phosphate buffer of pH 8.0, and its linear response was controlled in 0.05 -0.30 mM. LDH standards containing 2.0 mM PEG could exhibit long-term stability. Under the optimized conditions, a linear response for LDH from 50 to 1200 U L -1 and a detection limit of 31 U L -1 were obtained with good precision (RSD: 2.1 -2.2%, n = 10) and better recovery of 96 -105%. The method's characteristics was high sensitivity, low consumptions, simple operations, good precision and reliability, lending itself to the miniaturization of fluorophotometer which transformed into a bedside instrument in the hospital.

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Lactate biosensors: current status and outlook

et al. 2013

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Many research efforts over the last few decades have been devoted to sensing lactate as an important analytical target in clinical care, sport medicine, and food processing. Therefore, research in designing lactate sensors is no longer in its infancy and now is more directed toward viable sensors for direct applications. In this review, we provide an overview of the most immediate and relevant developments toward this end, and we discuss and assess common transduction approaches. Further, we critically describe the pros and cons of current commercial lactate sensors and envision how future sensing design may benefit from emerging new technologies.

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Determination of pyruvic acid by using enzymic fluorescence capillary analysis

Cited by 22 publications

References 21 publications

Rhodamine B Piperazinoacetohydrazine: A Water‐Soluble Spectroscopic Reagent for Pyruvic Acid Labeling

Rhodamine B Piperazinoacetohydrazine: A Water‐Soluble Spectroscopic Reagent for Pyruvic Acid Labeling

A Novel Analysis Method for Lactate Dehydrogenase Activity in Serum Samples Based on Fluorescence Capillary Analysis

Lactate biosensors: current status and outlook

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