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. 2023 May 29;28(11):4409.
doi: 10.3390/molecules28114409.

LC-MS Profiling of Kakkonto and Identification of Ephedrine as a Key Component for Its Anti-Glycation Activity

Kaori Ito  1 , Takashi Kikuchi  1 , Kanako Ikube  1 , Kouharu Otsuki  1 , Kazuo Koike  1 , Wei Li  1
Affiliations

LC-MS Profiling of Kakkonto and Identification of Ephedrine as a Key Component for Its Anti-Glycation Activity

Kaori Ito et al. Molecules. .

Abstract

A total of 147 oral Kampo prescriptions, which are used clinically in Japan, were evaluated for their anti-glycation activity. Kakkonto demonstrated significant anti-glycation activity, prompting further analysis of its chemical constituents using LC-MS, which revealed the presence of two alkaloids, fourteen flavonoids, two but-2-enolides, five monoterpenoids, and four triterpenoid glycosides. To identify the components responsible for its anti-glycation activity, the Kakkonto extract was reacted with glyceraldehyde (GA) or methylglyoxal (MGO) and analyzed using LC-MS. In LC-MS analysis of Kakkonto reacted with GA, the peak intensity of ephedrine was attenuated, and three products from ephedrine-scavenging GA were detected. Similarly, LC-MS analysis of Kakkonto reacted with MGO revealed two products from ephedrine reacting with MGO. These results indicated that ephedrine was responsible for the observed anti-glycation activity of Kakkonto. Ephedrae herba extract, which contains ephedrine, also showed strong anti-glycation activity, further supporting ephedrine's contribution to Kakkonto's reactive carbonyl species' scavenging ability and anti-glycation activity.

Keywords: Kakkonto; Kampo; advanced glycation end products; anti-glycation activity; ephedrine.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Anti-glycation activity of Kakkonto (A), Ephedrae herba extract (B), and aminoguanidine (positive control) (C) at different concentrations in the BSA–GA assay.
Figure 2
Figure 2
Total ion chromatograms of Kakkonto in positive ion mode (A) and negative ion mode (B).
Figure 3
Figure 3
Structures of identified compounds in Kakkonto.
Figure 4
Figure 4
MS/MS fragmentation pathway of compounds 1 and 2.
Figure 5
Figure 5
MS/MS fragmentation pathway of compounds 13, 18, and 20.
Figure 6
Figure 6
MS/MS fragmentation pathway of compounds 9, 10, and 17.
Figure 7
Figure 7
MS/MS fragmentation pathway of compounds 3, 4, 6, 7, 11, and 14.
Figure 8
Figure 8
MS/MS fragmentation pathway of compounds 12 and 16.
Figure 9
Figure 9
MS/MS fragmentation pathway of compounds 5, 8, 15, 22, and 23.
Figure 10
Figure 10
MS/MS fragmentation pathway of compounds 24–27.
Figure 11
Figure 11
The total ion chromatogram of Kakkonto reacted with glyceraldehyde (GA) and methyl glyoxal (MGO) in positive ion mode. (A) Superposition of Kakkonto extract reacted with GA (red) and without GA (black). (B) Superposition of Kakkonto extract reacted with MGO (red) and without MGO (black).
Figure 12
Figure 12
Reaction between ephedrine (1) and glyceraldehyde (GA), and MS/MS fragmentation pathway of the reaction products.
Figure 13
Figure 13
Reaction between ephedrine (1) and methylglyoxal (MGO), and MS/MS fragmentation pathway of the reaction products.
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References

    1. Welsh K.J., Kirkman M.S., Sacks D.B. Role of glycated proteins in the diagnosis and management of diabetes: Research gaps and future directions. Diabetes Care. 2016;39:1299–1306. doi: 10.2337/dc15-2727. - DOI - PMC - PubMed
    1. Lapolla A., Fedele D., Reitano R., Bonfante L., Guizzo M., Seraglia R., Tubaro M., Traldi P. Mass spectrometric study of in vivo production of advanced glycation endproducts/peptides. J. Mass Spectrom. 2005;40:969–972. doi: 10.1002/jms.842. - DOI - PubMed
    1. Lapolla A., Fedele D., Seraglia R., Traldi P. The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes: An update. Mass Spectrom. Rev. 2006;25:775–797. doi: 10.1002/mas.20090. - DOI - PubMed
    1. Zhang Q., Ames J.M., Smith R.D., Baynes J.W., Metz T.O. A Perspective on the maillard reaction and the analysis of protein glycation by mass spectrometry: Probing the pathogenesis of chronic disease. J. Proteome Res. 2009;8:754–769. doi: 10.1021/pr800858h. - DOI - PMC - PubMed
    1. Sjoblom N.M., Kelsey M.M.G., Scheck R.A. A Systematic study of selective protein glycation. Angew. Chem. Int. Ed. 2018;57:16077–16082. doi: 10.1002/anie.201810037. - DOI - PubMed

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