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  • Profiling proteins in nutraceutical formulations: characterization of the constituents.

Profiling proteins in nutraceutical formulations: characterization of the constituents.

Food chemistry (2015-10-17)
Alessia Bellomaria, Ridvan Nepravishta, Mario Marchetti, Maurizio Paci
ABSTRACT

Several nutraceutical preparations containing proteins, amino acids and other small molecules are nowadays present on the market. In this work we propose NMR spectroscopy such as (1)H NMR, (1)H-(1)H TOCSY and DOSY for their constituents characterization, identification and profiling, comparing these results with those obtained by electrophoretic technique such as SDS-PAGE. The (1)H NMR spectroscopy was applied for measurements of the amino acids and other small compounds added from the manufacturer. Further the autocorrelation function obtained from the one dimensional spectrum was used without the complete assignment of the resonances of the NMR spectrum of proteins for the evaluation of the folding quality and stability. Finally the DOSY NMR technique was performed on the samples for the characterization of the mean molecular weight range of proteins. All this features considered together create an important set of data useful for the evaluation of the protein profiling and the characterization of such formulations.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%
Sigma-Aldrich
Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Sodium hydrogencarbonate, JIS special grade, 99.5-100.3%
Sigma-Aldrich
Sodium hydrogencarbonate, SAJ first grade, 99.5-100.3%
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Sodium bicarbonate, ReagentPlus®, ≥99.5%, powder
Sigma-Aldrich
Sodium bicarbonate, ACS reagent, ≥99.7%
Sigma-Aldrich
Sodium bicarbonate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium bicarbonate, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E500, 99.0-100.5%, powder
Sigma-Aldrich
Sodium hydrogencarbonate, −40-+140 mesh, ≥95%
Sigma-Aldrich
Sodium bicarbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.7%
Sigma-Aldrich
L-Isoleucine, 99%, FCC, FG
Sigma-Aldrich
L-Isoleucine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
Taurine, BioUltra, ≥99.5% (T)
SAFC
Taurine
Sigma-Aldrich
Sodium bicarbonate-12C, 99.9 atom % 12C
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Taurine, suitable for cell culture, meets USP testing specifications
Sigma-Aldrich
Taurine, ≥99%
Sigma-Aldrich
L-Isoleucine, reagent grade, ≥98% (HPLC)
Supelco
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Sigma-Aldrich
L-Isoleucine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
SAFC
L-Isoleucine
Sigma-Aldrich
Taurine, ≥98%, FG