Development of a direct LC-ESI-MS method for the measurement of human serum carnosinase activity

Gilardoni, Ettore; Gervasoni, Silvia; Maspero, Marco; Dallanoce, Clelia; Vistoli, Giulio; Carini, Marina; Regazzoni, Luca

doi:10.1016/j.jpba.2020.113440

Cited by 15 publications

(38 citation statements)

References 28 publications

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“…The reason for these low to undetectable CAR concentrations is the presence of a highly active, HCD hydrolyzing, enzyme in human circulation, namely serum carnosinase (CN1). It is known that the affinity of CN1 toward ANS is, in comparison to CAR, lower due to the methylation of the imidazole-ring, resulting in a delay in breakdown of the dipeptide upon oral ingestion [ 8–12 ]. Indeed, an in vivo pharmacokinetic experiment showed that the combined ingestion of both dipeptides increases plasma accumulation of ANS beyond the level measured following ANS ingestion without CAR [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

The ergogenic effect of acute carnosine and anserine supplementation: dosing, timing, and underlying mechanism

Jager

Blancquaert

Stede

et al. 2022

Journal of the International Society of Sports Nutrition

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Background Recent studies suggest that acute-combined carnosine and anserine supplementation has the potential to improve the performance of certain cycling protocols. Yet, data on optimal dose, timing of ingestion, effective exercise range, and mode of action are lacking. Three studies were conducted to establish dosing and timing guidelines concerning carnosine and anserine intake and to unravel the mechanism underlying the ergogenic effects. Methods First, a dose response study A was conducted in which 11 men randomly received placebo, 10, 20, or 30 mg.kg −1 of both carnosine and anserine. They performed 3x maximal voluntary isometric contractions (MVC), followed by a 5 x 6 s repeated cycling sprint ability test (RSA), once before the supplement and 30 and 60 minutes after. In a second study, 15 men performed 3x MVCs with femoral nerve electrical stimulation, followed by an RSA test, once before 30 mg.kg −1 carnosine and anserine and 60 minutes after. Finally, in study C, eight men performed a high intensity cycling training after randomly ingesting 30 mg.kg −1 of carnosine and anserine, a placebo or antihistamines (reduce post-exercise blood flow) to investigate effects on muscle perfusion. Results Study A showed a 3% peak power (p = 0.0005; 95% CI = 0.07 to 0.27; ES = 0.91) and 4.5% peak torque (p = 0.0006; 95% CI = 0.12 to 0.50; ES = 0.87) improvement on RSA and MVC, with 30 mg.kg −1 carnosine + anserine ingestion 60 minutes before the performance yielding the best results. Study B found no performance improvement on group level; however, a negative correlation (r = −0.54; p = 0.0053; 95% CI = −0.77 to −0.19) was found between carnosinase enzyme activity (responsible for carnosine and anserine breakdown) and performance improvement. No effect of the supplement on neuromuscular function nor on muscle perfusion was found. Conclusions These studies reveal that acute ingestion of 30 mg.kg −1 of both carnosine and anserine, 60 minutes before a high intensity exercise, can potentially improve performance, such as short cycling sprints or maximal muscle contractions. Subjects with lower carnosinase activity, and thus a slower breakdown of circulating dipeptides, appear to benefit more from this ergogenic effect. Finally, neither the involvement of a direct effect on neuromuscular function, nor an indirect effect on recovery through increased muscle perfusion could be confirmed as potential mechanism of action. The ergogenic mechanism therefore remains elusive.

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Section: Introductionmentioning

confidence: 99%

The ergogenic effect of acute carnosine and anserine supplementation: dosing, timing, and underlying mechanism

Jager

Blancquaert

Stede

et al. 2022

Journal of the International Society of Sports Nutrition

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“…Unfortunately, considering administering carnosine systemically or orally the concern has to be raised that this may result in very low concentrations of the dipeptide in the target tissue. This has to be assumed, because of dipeptidase activities in the intestine [ 23 ] and in serum, where the dipeptide is rapidly cleaved by serum carnosinase (CN1) [ 24 ] with activity between 1.33 ± 0.08 nmol/h/µL [ 25 ] and 3.2 ± 1.1 μmol/mL/h [ 26 ]. On the other hand, several studies report beneficial effects of an oral administration of carnosine in a number of different diseases—some of them associated with neurological dysfunctions.…”

Section: Introductionmentioning

confidence: 99%

Erythrocytes Prevent Degradation of Carnosine by Human Serum Carnosinase

Oppermann

Elsel

Birkemeyer

et al. 2021

IJMS

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The naturally occurring dipeptide carnosine (β-alanyl-l-histidine) has beneficial effects in different diseases. It is also frequently used as a food supplement to improve exercise performance and because of its anti-aging effects. Nevertheless, after oral ingestion, the dipeptide is not detectable in human serum because of rapid degradation by serum carnosinase. At the same time, intact carnosine is excreted in urine up to five hours after intake. Therefore, an unknown compartment protecting the dipeptide from degradation has long been hypothesized. Considering that erythrocytes may constitute this compartment, we investigated the uptake and intracellular amounts of carnosine in human erythrocytes cultivated in the presence of the dipeptide and human serum using liquid chromatography–mass spectrometry. In addition, we studied carnosine’s effect on ATP production in red blood cells and on their response to oxidative stress. Our experiments revealed uptake of carnosine into erythrocytes and protection from carnosinase degradation. In addition, no negative effect on ATP production or defense against oxidative stress was observed. In conclusion, our results for the first time demonstrate that erythrocytes can take up carnosine, and, most importantly, thereby prevent its degradation by human serum carnosinase.

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“…By following this method, in a previous work we reported the synthesis of L-carnosine-D3 (L-1-[D3]), which was obtained from a trideuterated analog of L-histidine (L-4-[D3]) (Fig. 2a) [29]. However, although the successful result, by applying such histidine-D3-based approach the preparation of labeled D-stereoisomers of the dipeptide is not feasible and the synthesis of methylated analogs appears to be inefficient and expensive.…”

Section: Design and Synthesis Of 13 C Labeled Carnosine Analogsmentioning

confidence: 99%

“…The use of a stable isotope of carnosine as internal standard for HPLC-MS analysis of carnosine in biospecimens has been recently reported by our group [29]. Although it still retains drawbacks such as deuterium exchange in the synthesis process and metabolic instability, L-carnosine-D3 (L-1-…”

Section: Carnosine and Anserine Quantitation In Muscle Tissuementioning

confidence: 99%

“…In a previous work we have prepared a trideuterated analog of carnosine (L-carnosine-D3, L-1-[D3]) (Fig. 2a), which was successfully applied in an HPLC-MS method for the measurement of carnosinase activity in human serum [29]. Despite the result, deuterated standards present disadvantages that can limit the accuracy in quantitation by HPLC-MS.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis and characterization of 13C labeled carnosine derivatives for isotope dilution mass spectrometry measurements in biological matrices

Maspero

Gilardoni

Bonfanti

et al. 2021

Talanta

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Development of a direct LC-ESI-MS method for the measurement of human serum carnosinase activity

Cited by 15 publications

References 28 publications

The ergogenic effect of acute carnosine and anserine supplementation: dosing, timing, and underlying mechanism

The ergogenic effect of acute carnosine and anserine supplementation: dosing, timing, and underlying mechanism

Erythrocytes Prevent Degradation of Carnosine by Human Serum Carnosinase

Synthesis and characterization of 13C labeled carnosine derivatives for isotope dilution mass spectrometry measurements in biological matrices

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