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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| Format: | Article |
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MDPI AG
2021-11-01
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| Online Access: | https://www.mdpi.com/2297-8739/8/11/219 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T05:03:48Z |
| format | Article |
| id | doaj.art-1ba6dc361f5d47329be907689e8039e7 |
| institution | Directory Open Access Journal |
| issn | 2297-8739 |
| language | English |
| last_indexed | 2024-03-10T05:03:48Z |
| publishDate | 2021-11-01 |
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| spelling | doaj.art-1ba6dc361f5d47329be907689e8039e72023-11-23T01:30:06ZengMDPI AGSeparations2297-87392021-11-0181121910.3390/separations8110219Liquid Chromatography-Tandem Mass Spectrometry for Detecting Myosin Light Chain 3 in Dry-Aged BeefHeeyoung Lee0Yoonji Heo1Jong-Chan Kim2You-Shin Shim3Food Standard Research Center, Korea Food Research Institute, Wanju 55365, KoreaTechnical Assistance Center, Korea Food Research Institute, Wanju 55365, KoreaFood Standard Research Center, Korea Food Research Institute, Wanju 55365, KoreaFood Standard Research Center, Korea Food Research Institute, Wanju 55365, KoreaLiquid 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.https://www.mdpi.com/2297-8739/8/11/219myosin light chain 1liquid chromatography-tandem mass spectrometryfood qualitydry-aged beeftenderness |
| spellingShingle | 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 Separations myosin light chain 1 liquid chromatography-tandem mass spectrometry food quality dry-aged beef tenderness |
| title | 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_short | 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 |
| topic | myosin light chain 1 liquid chromatography-tandem mass spectrometry food quality dry-aged beef tenderness |
| url | https://www.mdpi.com/2297-8739/8/11/219 |
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