Quantification of bovine α-lactalbumin in infant milk formula using LC-MS

Kleinnijenhuis, Anne J.; Gool, Martine P. van; Holthoon, Frédérique L. van; Noort, Maarten van den; Huppertz, Thom

doi:10.1016/j.idairyj.2020.104899

Cited by 2 publications

(5 citation statements)

References 52 publications

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“…In recently published studies selected marker peptides have been used for quantifying β-LG and α-LA in various milk products, such as raw milk, milk powder, infant formulas, and fresh cheese [24][25][26][27][28][29]. For example, the marker peptide TPEVDDEALEK has been used to quantify β-LG in milk powder [24] and fresh cheese [27].…”

Section: Quantification Of α-Lactalbumin and β-Lactoglobulin In Model...mentioning

confidence: 99%

“…For example, the marker peptide TPEVDDEALEK has been used to quantify β-LG in milk powder [24] and fresh cheese [27]. The peptides VGINYWLAHK, LDQWLcEK, and cEVFR have been used to quantify α-LA in raw milk [25] and infant milk formula [26]. Based on a bottom-up proteomic approach using isotopically labeled peptides as internal standards, Bär et al [28] presented a multiple-reaction monitoring liquid chromatography-mass spectrometry (MRM-LC-MS/MS) method that enables the simultaneous quantification of twenty milk proteins in various dairy products, e.g., in Swiss Emmental (which is a raw milk cheese).…”

Section: Quantification Of α-Lactalbumin and β-Lactoglobulin In Model...mentioning

confidence: 99%

“…Specific marker peptides for individual milk proteins were used for species identification [19,20] and allergen detection [21,22]. Furthermore, studies regarding the quantification of individual milk proteins in milk [23], milk powder [24], infant formulas [25,26], or fresh cheese [27] were published. Moreover, an initial bottom-up approach for the quantification of milk proteins in various milk products, including Emmental-type cheese, was presented [28].…”

Section: Introductionmentioning

confidence: 99%

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Identification of Marker Peptides for the Whey Protein Quantification in Edam-Type Cheese

Oesen

Treblin

Clawin‐Rädecker

et al. 2023

Foods

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Several technologies are available for incorporating whey proteins into a cheese matrix. However, there is no valid analytical method available to determine the whey protein content in matured cheese, to date. Consequently, the aim of the present study was to develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of individual whey proteins based on specific marker peptides (‘bottom-up’ proteomic approach). Therefore, the whey protein-enriched model of the Edam-type cheese was produced in a pilot plant and on an industrial scale. Tryptic hydrolysis experiments were performed to evaluate the suitability of identified potential marker peptides (PMPs) for α-lactalbumin (α-LA) and β-lactoglobulin (β-LG). Based on the findings, α-LA and β-LG appeared to be resistant to proteolytic degradation during six weeks of ripening and no influence on the PMP was observed. Good levels of linearity (R2 > 0.9714), repeatability (CVs < 5%), and recovery rate (80% to 120%) were determined for most PMPs. However, absolute quantification with external peptide and protein standards revealed differences in model cheese depending on the PMP, e.g., 0.50% ± 0.02% to 5.31% ± 0.25% for β-LG. As protein spiking prior to hydrolysis revealed differing digestion behavior of whey proteins, further studies are required to enable valid quantification in various cheese types.

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Section: Quantification Of α-Lactalbumin and β-Lactoglobulin In Model...mentioning

confidence: 99%

Section: Quantification Of α-Lactalbumin and β-Lactoglobulin In Model...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification of Marker Peptides for the Whey Protein Quantification in Edam-Type Cheese

Oesen

Treblin

Clawin‐Rädecker

et al. 2023

Foods

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“…10,11 Furosine, formed by the hydrolysis of Amadori compounds, is usually detected by high-pressure liquid chromatography (HPLC), 12 ultra-pressure liquid chromatography (UPLC), or gas chromatography (GC) combined with fluorescence detection or mass spectrometry (MS). 7,13 Lactosylation sites can be analyzed directly by MS 14 or MS coupled with separation techniques, such as liquid chromatography (LC), 2,4,6,15,16 two-dimensional (2D) gel electrophoresis, 17,18 or capillary electrophoresis (CE). 3,19 The immunoassay techniques 12,19 are also applied.…”

Section: Introductionmentioning

confidence: 99%

“…Commercially available boronate-affinity materials (BAMs) are used to enrich samples in lactosylated peptides, , which allows the detection of low-concentration components of samples containing sugar moieties. Recently, Kijewska and co-workers have developed a functionalized resinPhB-Lys(PhB)-ChemMatrix Rink resinthat selectively and efficiently captures glycated peptides. , Furthermore, tags containing quaternary ammonium salt and phenylboronic acid derivatives, appropriately designed and synthesized, were used to increase the ionization efficiency of glycoconjugates in mass spectrometry analysis. , Furosine, formed by the hydrolysis of Amadori compounds, is usually detected by high-pressure liquid chromatography (HPLC), ultra-pressure liquid chromatography (UPLC), or gas chromatography (GC) combined with fluorescence detection or mass spectrometry (MS). , Lactosylation sites can be analyzed directly by MS or MS coupled with separation techniques, such as liquid chromatography (LC), ,,,, two-dimensional (2D) gel electrophoresis, , or capillary electrophoresis (CE). , The immunoassay techniques , are also applied. The advantage of mass spectrometry is its high sensitivity and specificity, which allows the identification and quantification of protein modification products.…”

Section: Introductionmentioning

confidence: 99%

High-Temperature, Solid-Phase Reaction of α-Amino Groups in Peptides with Lactose and Glucose: An Alternative Mechanism Leading to an α-Ketoacyl Derivative

Kijewska

Zawadzka

Stefanowicz

2023

J. Agric. Food Chem.

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The reaction of proteins with reducing sugars results in the formation of Amadori products, which involves the Nterminal group and/or ε-amino group of the lysine side chain. However, less attention has been given to the reactivity of the Nterminus of a peptide chain under similar conditions. In our work, we focused on the reaction of the α-amino group of peptides in the presence of a reducing sugar, specifically lactose. We optimized the reaction conditions of model peptides with lactose in the solid phase and showed that temperatures above 120 °C lead to the deamination of the N-terminal amino acid moiety, ultimately resulting in α-ketoacids. We carried out detailed studies to confirm the structure of the deaminated product using analytical methods such as ESI-MS and LC−MS/MS, as well as chemical methods that involved the reduction of the carbonyl group combined with isotopic exchange and the reactivity of the carbonyl group with the hydroxylamine derivative. The structure of the reaction product was also confirmed by chemical synthesis. We suggested plausible mechanisms for the formation of the deaminated product and considered the probable path of its formation. Our aim was to determine whether the reaction proceeds according to the Streckerbased mechanism and direct imine isomerization by carrying out reactions of model peptides in the presence of lactose under aerobic and anaerobic conditions and comparing the results obtained. KEYWORDS: LC−MS/MS analysis, lactosylated peptides, α-ketoacid derivatives, deamination catalyzed by a reducing sugar ■ HIGHLIGHTS• LC−MS/MS analysis of lactosylated peptides.• Deamination of the N-terminal amino acid by a reducing sugar. • Chemoselective reaction of the hydroxylamine derivative with the deamination product.

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Quantification of bovine α-lactalbumin in infant milk formula using LC-MS

Cited by 2 publications

References 52 publications

Identification of Marker Peptides for the Whey Protein Quantification in Edam-Type Cheese

Identification of Marker Peptides for the Whey Protein Quantification in Edam-Type Cheese

High-Temperature, Solid-Phase Reaction of α-Amino Groups in Peptides with Lactose and Glucose: An Alternative Mechanism Leading to an α-Ketoacyl Derivative

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