Autofluorescence-Based Investigation of Spatial Distribution of Phenolic Compounds in Soybeans Using Confocal Laser Microscopy and a High-Resolution Mass Spectrometric Approach

Mayya P Razgonova; Yulia N Zinchenko; Darya K Kozak; Victoria A Kuznetsova; Alexander M Zakharenko; Sezai Ercisli; Kirill S Golokhvast

doi:10.3390/molecules27238228

Autofluorescence-Based Investigation of Spatial Distribution of Phenolic Compounds in Soybeans Using Confocal Laser Microscopy and a High-Resolution Mass Spectrometric Approach

Molecules. 2022 Nov 25;27(23):8228. doi: 10.3390/molecules27238228.

Authors

Mayya P Razgonova^{1

2}, Yulia N Zinchenko^{1

2}, Darya K Kozak³, Victoria A Kuznetsova^{1

3}, Alexander M Zakharenko⁴, Sezai Ercisli⁵, Kirill S Golokhvast^{1

2

4}

Affiliations

¹ Far Eastern Experimental Station, N.I. Vavilov All-Russian Institute of Plant Genetic Resources, 190000 Saint-Petersburg, Russia.
² SEC Nanotechnology, Polytechnic Institute, Far Eastern Federal University, 690922 Vladivostok, Russia.
³ Laboratory of Biochemistry, Blagoveshchensk State Pedagogical University, 675000 Blagoveshchensk, Russia.
⁴ Laboratory of Pesticide Toxicology, Siberian Federal Scientific Center of Agrobiotechnology RAS, 633501 Krasnoobsk, Russia.
⁵ Department of Horticulture, Agricultural Faculty, Ataturk University, Erzurum 25240, Turkey.

Abstract

In this research, we present a detailed comparative analysis of the bioactive substances of soybean varieties k-11538 (Russia), k-11559 (Russia), k-569 (China), k-5367 (China), k-5373 (China), k-5586 (Sweden), and Primorskaya-86 (Russia) using an LSM 800 confocal laser microscope and an amaZon ion trap SL mass spectrometer. Laser microscopy made it possible to clarify in detail the spatial arrangement of the polyphenolic content of soybeans. Our results revealed that the phenolics of soybean are spatially located mainly in the seed coat and the outer layer of the cotyledon. High-performance liquid chromatography (HPLC) was used in combination with an amaZon SL BRUKER DALTONIKS ion trap (tandem mass spectrometry) to identify target analytes in soybean extracts. The results of initial studies revealed the presence of 63 compounds, and 45 of the target analytes were identified as polyphenolic compounds.

Keywords: Glycine Willd; HPLC-MS/MS; flavonols; laser microscopy; polyphenols; tandem mass spectrometry.

MeSH terms

Chromatography, High Pressure Liquid
Glycine max* / chemistry
Lasers
Microscopy, Confocal
Phenols* / chemistry
Plant Extracts / chemistry
Tandem Mass Spectrometry / methods

Substances

Phenols
Plant Extracts

Grants and funding

0662-2019-0006/Ministry of Science and High Education of the Russian Federation