Simultaneous determination of lactate and pyruvate in human sweat using reversed‐phase high‐performance liquid chromatography: a noninvasive approach

Biagi, Simona; Ghimenti, Silvia; Onor, Massimo; Bramanti, Emilia

doi:10.1002/bmc.2713

Cited by 79 publications

(49 citation statements)

References 47 publications

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“…For lactate determination the dynamic linear range (DLR) was 0.03–350 mM, 0.03 mM LOD, slope 47.9 ± 0.62 (au mM -1 ), R 2 = 0.9987. The figures of merit were comparable with those obtained previously [ 47 ]. The coefficient of variation (CV%) of measurements performed on the same vial was < 1%; the coefficient of variation (CV%) of measurements performed on different vials was < 2%.…”

Section: Methodssupporting

confidence: 89%

Thallium stimulates ethanol production in immortalized hippocampal neurons

et al. 2017

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Lactate and ethanol (EtOH) were determined in cell culture medium (CCM) of immortalized hippocampal neurons (HN9.10e cell line) before and after incubation with Thallium (Tl). This cell line is a reliable, in vitro model of one of the most vulnerable regions of central nervous system. Cells were incubated for 48 h with three different single Tl doses: 1, 10, 100 μg/L (corresponding to 4.9, 49 and 490 nM, respectively). After 48 h, neurons were “reperfused” with fresh CCM every 24/48 h until 7 days after the treatment and the removed CCM was collected and analysed. Confocal microscopy was employed to observe morphological changes. EtOH was determined by head space—solid phase microextraction -gas chromatography -mass spectrometry (HS-SPME-GCMS), lactate by RP-HPLC with UV detection. Tl exposure had significant effects on neuronal growth rate and morphology. The damage degree was dose-dependent. In not exposed cells, EtOH concentration was 0.18 ± 0.013 mM, which represents about 5% of lactate concentration (3.4 ± 0.10 mM). After Tl exposure lactate and EtOH increased. In CCM of 100 and 10 μg/L Tl-treated cells, lactate increased 24 h after reperfusion up to 2 and 3.3 times the control value, respectively. In CCM of 10 and 100 μg/L Tl-treated cells 24 h after reperfusion, EtOH increased up to 0.3 and 0.58 mmol/L. respectively. These results are consistent with significant alterations in energy metabolism, despite the low doses of Tl employed and the relatively short incubation time.

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Section: Methodssupporting

confidence: 89%

Thallium stimulates ethanol production in immortalized hippocampal neurons

et al. 2017

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“…The color changes occurred on timescales of <1 min (movie S4). The concentration of lactate in sweat is an indicator of exercise intolerance and tissue hypoxia (44, 45). Enzymatic reactions between lactate and co-factor NAD + (nicotinamide adenine dinucleotide) by lactate dehydrogenase and diaphorase induce a change in color of a chromogenic reagent (that is, formazan dyes).…”

Section: Resultsmentioning

confidence: 99%

A soft, wearable microfluidic device for the capture, storage, and colorimetric sensing of sweat

et al. 2016

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Capabilities in health monitoring via capture and quantitative chemical analysis of sweat could complement, or potentially obviate the need for, approaches based on sporadic assessment of blood samples. Established sweat monitoring technologies use simple fabric swatches and are limited to basic analysis in controlled laboratory or hospital settings. We present a collection of materials and device designs for soft, flexible and stretchable microfluidic systems, including embodiments that integrate wireless communication electronics, which can intimately and robustly bond to the surface of skin without chemical and mechanical irritation. This integration defines access points for a small set of sweat glands such that perspiration spontaneously initiates routing of sweat through a microfluidic network and set of reservoirs. Embedded chemical analyses respond in colorimetric fashion to markers such as chloride and hydronium ions, glucose and lactate. Wireless interfaces to digital image capture hardware serve as a means for quantitation. Human studies demonstrated the functionality of this microfluidic device during fitness cycling in a controlled environment and during long-distance bicycle racing in arid, outdoor conditions. The results include quantitative values for sweat rate, total sweat loss, pH and concentration of both chloride and lactate.

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“…Pyruvate, lactate NAD + , and NADH were quantified by HPLC according to the published procedure51 with some modifications. An HPLC gradient pump (L-2310, Hitachi, Tokyo, Japan) was coupled with a Rheodyne 7125 injector equipped with a 20 μL sample loop (Rheodyne, Cotati, CA, USA), an L-4000 UV detector (Hitachi), and a 655A-52 column oven (Hitachi).…”

Section: Methodsmentioning

confidence: 99%

Identification of lactate dehydrogenase as a mammalian pyrroloquinoline quinone (PQQ)-binding protein

Akagawa

Minematsu

Shibata

et al. 2016

Sci Rep

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Pyrroloquinoline quinone (PQQ), a redox-active o-quinone, is an important nutrient involved in numerous physiological and biochemical processes in mammals. Despite such beneficial functions, the underlying molecular mechanisms remain to be established. In the present study, using PQQ-immobilized Sepharose beads as a probe, we examined the presence of protein(s) that are capable of binding PQQ in mouse NIH/3T3 fibroblasts and identified five cellular proteins, including l-lactate dehydrogenase (LDH) A chain, as potential mammalian PQQ-binding proteins. In vitro studies using a purified rabbit muscle LDH show that PQQ inhibits the formation of lactate from pyruvate in the presence of NADH (forward reaction), whereas it enhances the conversion of lactate to pyruvate in the presence of NAD+ (reverse reaction). The molecular mechanism underlying PQQ-mediated regulation of LDH activity is attributed to the oxidation of NADH to NAD+ by PQQ. Indeed, the PQQ-bound LDH oxidizes NADH, generating NAD+, and significantly catalyzes the conversion of lactate to pyruvate. Furthermore, PQQ attenuates cellular lactate release and increases intracellular ATP levels in the NIH/3T3 fibroblasts. Our results suggest that PQQ, modulating LDH activity to facilitate pyruvate formation through its redox-cycling activity, may be involved in the enhanced energy production via mitochondrial TCA cycle and oxidative phosphorylation.

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Simultaneous determination of lactate and pyruvate in human sweat using reversed‐phase high‐performance liquid chromatography: a noninvasive approach

Cited by 79 publications

References 47 publications

Thallium stimulates ethanol production in immortalized hippocampal neurons

Thallium stimulates ethanol production in immortalized hippocampal neurons

A soft, wearable microfluidic device for the capture, storage, and colorimetric sensing of sweat

Identification of lactate dehydrogenase as a mammalian pyrroloquinoline quinone (PQQ)-binding protein

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