Methods for Extraction of Muscle Proteins from Meat and Fish Using Denaturing and Nondenaturing Solutions

Malva, Antonella della; Albenzio, Marzia; Santillo, Antonella; Russo, Donatella Esterina; Figliola, Lucia; Caroprese, Mariangela; Marino, R.

doi:10.1155/2018/8478471

Cited by 27 publications

(18 citation statements)

References 20 publications

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“…In this study, samples of gastrocnemius muscle were subjected to three sequential extractions, each one being analyzed by LC-MS/MS. In particular, phosphate buffer saline (PBS) was used to solubilize hydrophilic proteins (PBS extract), and then the insoluble part was treated with a combination of urea (U) and thiourea (T) to extract hydrophobic molecules such as membrane proteins, myofibrillar, and part of ECM proteins (U/T extract) [ 31 ]. Finally, the remaining insoluble fraction was treated with guanidine-HCl (GuHCl extract), one of the most efficient chaotropic agents which is known to extract poorly soluble, heavily cross-linked proteins, and proteoglycans.…”

Section: Resultsmentioning

confidence: 99%

Age-Related Changes in the Matrisome of the Mouse Skeletal Muscle

Lofaro

Cisterna

Lacavalla

et al. 2021

IJMS

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Aging is characterized by a progressive decline of skeletal muscle (SM) mass and strength which may lead to sarcopenia in older persons. To date, a limited number of studies have been performed in the old SM looking at the whole, complex network of the extracellular matrix (i.e., matrisome) and its aging-associated changes. In this study, skeletal muscle proteins were isolated from whole gastrocnemius muscles of adult (12 mo.) and old (24 mo.) mice using three sequential extractions, each one analyzed by liquid chromatography with tandem mass spectrometry. Muscle sections were investigated using fluorescence- and transmission electron microscopy. This study provided the first characterization of the matrisome in the old SM demonstrating several statistically significantly increased matrisome proteins in the old vs. adult SM. Several proteomic findings were confirmed and expanded by morphological data. The current findings shed new light on the mutually cooperative interplay between cells and the extracellular environment in the aging SM. These data open the door for a better understanding of the mechanisms modulating myocellular behavior in aging (e.g., by altering mechano-sensing stimuli as well as signaling pathways) and their contribution to age-dependent muscle dysfunction.

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Section: Resultsmentioning

confidence: 99%

Age-Related Changes in the Matrisome of the Mouse Skeletal Muscle

Lofaro

Cisterna

Lacavalla

et al. 2021

IJMS

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“…The following general settings were used: curtain gas, 30 psi; ion source gas (1), 50 psi; ion source gas (2), 55 psi; temperature 550 °C; and ion spray voltage, 5500 V. We developed a precision analysis method using LC/MS/MS to measure the myl3 concentration in beef samples as a quality test for dry aging. Myofibrillar proteins, mainly myosin and actin, are the primary components of the skeletal muscle [17]. We hypothe- We developed a precision analysis method using LC/MS/MS to measure the myl3 concentration in beef samples as a quality test for dry aging.…”

Section: Resultsmentioning

confidence: 99%

“…We hypothe- We developed a precision analysis method using LC/MS/MS to measure the myl3 concentration in beef samples as a quality test for dry aging. Myofibrillar proteins, mainly myosin and actin, are the primary components of the skeletal muscle [17]. We hypothesized that myosin isolated from muscle skeletal protein is related to beef tenderness.…”

Section: Resultsmentioning

confidence: 99%

Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef

et al. 2021

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Liquid chromatography-tandem mass spectrometry (LC/MS/MS) is a more accurate technique for detecting proteins than electrophoresis-based methods such as western blotting. Because of its convenience, western blotting is commonly used for protein analysis in beef. We developed a method for detecting myosin light chain 3 (myl3) in beef samples, particularly dry-aged beef, using LC/MS/MS for quality testing. Musculus longissimus dorsi of Holstein was aged for 0, 2, 4, 5, 9, 11, 17, 20, and 24 weeks and used to measure the myl3 concentration. Because of the high molecular weight of myl3, the limitations of LC/MS/MS were overcome by implementing immunoprecipitation and digestion steps. Ultimately, a tryptic fragment of myl3 (13-mer), generated using immunoprecipitation and digestion by a biotinylated antibody, was detected using LC-MS/MS in positive ion mode through multiple reaction monitoring and analyte separation on a C18 column. Our method showed limits of detection and quantification of less than 0.3 and 0.8 μg/kg, respectively. However, differences in the myl3 concentrations according to the aging time were not significant (p > 0.05). After 12 weeks, myl3 disappeared in tested all samples, thus our analytical method can be used for accurate measurement of muscle protein in beef samples.

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“…This is particularly true as the samples with the highest concentrations: T080 (20% w/w chicken) and B080 (20% w/w pork) always gave the best classification accuracy whereas, those with the lowest concentration T099 (1% w / w chicken) and B099 (1% w / w pork) consistently gave the worst. Extraction of meat compounds often involve using denaturing or nondenaturing solutions which, can be expensive [ 49 ]. As observed for the determination of optimal dilution, better accuracies could be achieved with this method for meat mixtures with higher adulterant ranges and when 1% w / w of mixture combination was not included.…”

Section: Discussionmentioning

confidence: 99%

Standardized Extraction Techniques for Meat Analysis with the Electronic Tongue: A Case Study of Poultry and Red Meat Adulteration

Zaukuu

Gillay

Kovács

2021

Sensors

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The electronic tongue (e-tongue) is an advanced sensor-based device capable of detecting low concentration differences in solutions. It could have unparalleled advantages for meat quality control, but the challenges of standardized meat extraction methods represent a backdrop that has led to its scanty application in the meat industry. This study aimed to determine the optimal dilution level of meat extract for e-tongue evaluations and also to develop three standardized meat extraction methods. For practicality, the developed methods were applied to detect low levels of meat adulteration using beef and pork mixtures and turkey and chicken mixtures as case studies. Dilution factor of 1% w/v of liquid meat extract was determined to be the optimum for discriminating 1% w/w, 3% w/w, 5% w/w, 10% w/w, and 20% w/w chicken in turkey and pork in beef with linear discriminant analysis accuracies (LDA) of 78.13% (recognition) and 64.73% (validation). Even higher LDA accuracies of 89.62% (recognition) and 68.77% (validation) were achieved for discriminating 1% w/w, 3% w/w, 5% w/w, 10% w/w, and 20% w/w of pork in beef. Partial least square models could predict both sets of meat mixtures with good accuracies. Extraction by cooking was the best method for discriminating meat mixtures and can be applied for meat quality evaluations with the e-tongue.

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Methods for Extraction of Muscle Proteins from Meat and Fish Using Denaturing and Nondenaturing Solutions

Cited by 27 publications

References 20 publications

Age-Related Changes in the Matrisome of the Mouse Skeletal Muscle

Age-Related Changes in the Matrisome of the Mouse Skeletal Muscle

Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef

Standardized Extraction Techniques for Meat Analysis with the Electronic Tongue: A Case Study of Poultry and Red Meat Adulteration

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