MG-H1 Quantification in Serum by LC-MS/MS

MG-H1 contents in the serum was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) as described previously.^{(16 (link))} Briefly, low molecular weight fractions (<3,000) of serum (50 µl) were obtained by VIVASPIN 500 (Sartorius Stedim Biotech, Goettingen, Germany), and reduced by NaBH₄. Standard [²H₂] MG-H1 (PolyPeptide Laboratories, Strasbourg, France) and [¹³C₆] Lysine (Cambridge Isotope Laboratories, Inc., Tewksbury, MA) were added to the reduced fractions. The sample was passed over a Strata-X-C column (Phenomenex, Torrance, CA) and assayed by LC-MS/MS using a TSQ Vantage triple stage quadrupole mass spectrometer (Thermo Fisher Scientific, Waltham, MA). The retention time for MG-H1 and Lysine were approximately 12 and 13 min, respectively. MG-H1, Lysine, and the standard were detected by electrospray positive ionization-mass spectrometric multiple reaction monitoring.

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Yamanaka M., Matsumura T., Ohno R.I., Fujiwara Y., Shinagawa M., Sugawa H., Hatano K., Shirakawa J.I., Kinoshita H., Ito K., Sakata N., Araki E, & Nagai R. (2016). Non-invasive measurement of skin autofluorescence to evaluate diabetic complications. Journal of Clinical Biochemistry and Nutrition, 58(2), 135-140.

Publication 2016

VIVASPIN 500 [13C6] Lysine Strata-X-C column TSQ Vantage triple stage quadrupole mass spectrometer

Electrospray ionization mass spectrometric Isotope Liquid chromatography Lysine Polypeptide Retention Serum Tandem mass spectrometry

Corresponding Organization :

Other organizations : Tokai University, Kumamoto University, Fukuoka University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

MG-H1 contents in the serum

control variables

Serum sample volume (50 µl)
Molecular weight fractions (<3,000)
Standard [2H2] MG-H1 and [13C6] Lysine addition
Strata-X-C column used for sample preparation
LC-MS/MS method with TSQ Vantage triple stage quadrupole mass spectrometer
Retention time for MG-H1 and Lysine (approximately 12 and 13 min, respectively)

controls

Positive control: Standard [2H2] MG-H1 and [13C6] Lysine
Negative control: Not explicitly mentioned

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