Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef
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 m...
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2021
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oai:doaj.org-article:1ba6dc361f5d47329be907689e8039e72021-11-25T18:59:27ZLiquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef10.3390/separations81102192297-8739https://doaj.org/article/1ba6dc361f5d47329be907689e8039e72021-11-01T00:00:00Zhttps://www.mdpi.com/2297-8739/8/11/219https://doaj.org/toc/2297-8739Liquid 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. <i>Musculus longissimus dorsi</i> 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 (<i>p</i> > 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.Heeyoung LeeYoonji HeoJong-Chan KimYou-Shin ShimMDPI AGarticlemyosin light chain 1liquid chromatography-tandem mass spectrometryfood qualitydry-aged beeftendernessPhysicsQC1-999ChemistryQD1-999ENSeparations, Vol 8, Iss 219, p 219 (2021) |
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DOAJ |
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myosin light chain 1 liquid chromatography-tandem mass spectrometry food quality dry-aged beef tenderness Physics QC1-999 Chemistry QD1-999 |
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myosin light chain 1 liquid chromatography-tandem mass spectrometry food quality dry-aged beef tenderness Physics QC1-999 Chemistry QD1-999 Heeyoung Lee Yoonji Heo Jong-Chan Kim You-Shin Shim Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| description |
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. <i>Musculus longissimus dorsi</i> 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 (<i>p</i> > 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. |
| format |
article |
| author |
Heeyoung Lee Yoonji Heo Jong-Chan Kim You-Shin Shim |
| author_facet |
Heeyoung Lee Yoonji Heo Jong-Chan Kim You-Shin Shim |
| author_sort |
Heeyoung Lee |
| title |
Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| title_short |
Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| title_full |
Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| title_fullStr |
Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| title_full_unstemmed |
Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged Beef |
| title_sort |
liquid chromatography-tandem mass spectrometry for detecting myosin light chain 3 in dry-aged beef |
| publisher |
MDPI AG |
| publishDate |
2021 |
| url |
https://doaj.org/article/1ba6dc361f5d47329be907689e8039e7 |
| work_keys_str_mv |
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