Development of a Method for Producing Purified Spinach Extract with High Content of 20-Hydroxyecdysone and Polyphenols

Authors

  • S. N. Zorin Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation
  • N. A. Petrov Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation
  • I. B. Perova Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation
  • A. D. Malinkin Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation
  • D. O. Bokov Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation
  • V. V. Bessonov Federal Research Centre of Nutrition, Biotechnology and Food Safety, Moscow, 109240, Russian Federation

Abstract

Adaptogens are biologically active substances providing state unspecific increased resistance in stressful situations. Spinach (Spinacia oleracea L.) is one of the most promising food plants containing adaptogens (ecdysteroids and polyphenols complexes). This research aims to obtain the final purified spinach extract (FPSE) with a high content of 20-hydroxyecdysone (20E), flavonoids, and oxalic acid-free from food raw materials – spinach leaves. This work is necessary to further use spinach as part of functional food ingredients in specialized food products. Total polyphenols were determined by the Folin-Ciocalteu method, oxalic acid content – by permanganate titration. The 20E content was determined by HPLC-MS using an Agilent 1100 chromatograph with an Agilent 6410 mass detector. Individual flavonoid content and profile were determined using an ultimate 3000 liquid chromatography system with a diode array detector (DAD) and a TSQ Endura triple quadrupole mass spectrometric detector (MSD). FPSE was obtained by water extraction of freeze-dried powdered spinach leaves (FDPSL), ultrafiltration, and sorption on a C18 column. In the FPSE, the 20E content was 12,17 ± 1,24 mg/g; total polyphenols – 19.3 ± 1.6%; flavonoids – 41.9 ± 4.1%, oxalic acid was absent. Flavonoid profile included patuletin-3-glucosyl-(1→6)-apiosyl-(1→2)-glucoside, patuletin-3-glucosyl-(1→6)-glucoside, patuletin-3-(2”feruloylglucosyl)-(1→6)-apiosyl-(1→2)-glucoside, patuletin-3-(2"feruloylglucosyl)-(1→6)-glucoside, axilyarin-4'-glucuronide (spinatoside), 5,3',4'-trihydroxy-3-methoxy-6:7-methylenedioxyflavone-4'-glucuronide, 5,4′-dihydroxy-3-methoxy-6:7-methylenedioxyflavone-4'-β-D-glucuronide, 5,4'-dihydroxy-3,3'-dimethoxy-6:7-methylenedioxyflavone-4'-glucuronide. As a result of the study, a laboratory method of producing functional food ingredient (adaptogen spinach extract) perspective for further industrial scaling and producing biologically active food supplements has been developed.

Keywords:

20-hydroxyecdysone, Flavonoids, Spinach, Spinacia oleracea L.

References

1. Jaremenko KV. Lazarev's theory of state unspecific increased resistance (SUIR) and adaptogens as a basis of preventive medicine. Psychopharmacology and Biological Narcology. 2005; 5(4): 1086-1092. Available from: https://clinpharm-journal.ru/articles/

2. Zabrodin ON. Conception of N.V. Lazarev about adaptogens in aspect of teaching about of nervous trophism. Psychopharmacology and Biological Narcology. 2005; 5(4): 1108-1112. Available from: https://clinpharm-journal.ru/articles/

3. Molinos DÁ. Effects of adaptogen supplementation on sport performance. A recent review of published studies. Journal of Human Sport and Exercise. 2013; 8(4): 1054-1066. Available from: doi.org/10.4100/jhse.2013.84.15

4. Volodin VV, Sidorova YuS, Mazo VK. 20-Hydroxyecdysone plant adaptogen: an anabolic effect, possible use in sports nutrition. Voprosy Pitaniia. 2013; 82(6): 24-30. Available from: https://www.voprosy-pitaniya.ru/en/jarticles_diet/233.html

5. Sidorova YuS, Selyaskin KE, Zorin SN, Vasilevskaya LS, Volodin VV, Mazo VK. In-vivo study of Serratula coronata L. extract on biomarkers of general adaptation syndrome. Traditional medicine 2014; 1 (36): 57-62. Available from: http://www.tradmed.ru/n36_9.shtml

6. Volodin VV, Pchelenko LD, Volodina SO, Kudryashova AG, Shevchenko OG, Zagorskaya NP. Pharmacological estimate of new containing ecdysteroid substance “Serpisten". Rastitel'nye resursy. 2006; 42(3): 113-130. Available from: https://www.elibrary.ru/item.asp?id=9275187

7. Kudyasheva AG, Andreeva LI, Volodin VV, Volodina SO. Biochemical parallels of cellular adaptive reactions in chronic low-intensity irradiation and the action of the phytoecdysteroid drug Serpisten. Radiation biology. Radioecology. 2015;55(1):43-50.

8. Shirshova TI, Politova NK, Beshlei IV, Volodin VV, Burtseva SA. Antimicrobial activity of natural ecdysteroids from Serratula coronata L. and their acyl derivatives. Pharmaceutical Chemistry Journal. 2006; 40(5): 268-271. Available from:

9. Selyaskin KE, Sidorova YS, Zorin SN, Volodin VV, Mazo VK. Effect of Serratula coronata extract on apoptosis activity in rats. Pharmaceutical Chemistry Journal. 2016; 50 (5): 315-319. Available from: doi.org/10.1007/s11094-006-0106-7

10. Odinokov VN, Galyautdinov IV, Nedopekin DV, Khalilov LM, Shashkov AS. Phytoecdysteroids from the juice of Serratula coronata L. (Asteraceae). Insect Biochemistry and Molecular Biology. 2002; 32(2): 161-165. Available from: 10.1016/S0965-1748(01)00106-0

11. Zharikov Ya A, Volodina SO, Volodin VV, Kaneva LA. Effect of infusion of Serratula coronata on the metabolism and growth young sheep. Russian Agricultural Sciences. 2019; 3: 51-53. Available from: doi.org/10.31857/S2500-26272019351-53

12. Bezmaternykh KV, Volodin VV, Volodina SO, Smirnova GV, Oktyabrsky ON. Study of antioxidant activity and adaptogenic activity of plant extracts containing ecdysteroids and polyphenols. In the book: Phenolic compounds: fundamental and applied aspects. Proceedings of the IX International Siposium. 2015; 499-502.

13. Bakrim A, Maria A, Sayah F, Lafont R, Takvorian N. Ecdysteroids in spinach (Spinacia oleracea L.): biosynthesis, transport and regulation of levels. Plant Physiology and Biochemistry. 2008; 46(10): 844-854. Available from: doi.org/10.1016/j.plaphy.2008.06.002

14. Edenharder R, Keller G, Platt K L, Unger K K. Isolation and characterization of structurally novel antimutagenic flavonoids from spinach (Spinacia oleracea). Journal of Agricultural and Food Chemistry. 2001; 49(6): 2767-2773. Available from: doi.org/10.1021/jf0013712

15. Koh E, Charoenprasert S, Mitchell AE. Effect of organic and conventional cropping systems on ascorbic acid, vitamin C, flavonoids, nitrate, and oxalate in 27 varieties of spinach (Spinacia oleracea L.). Journal of Agricultural and Food Chemistry. 2012; 60(12): 3144-3150. Available from: doi.org/10.1021/jf300051f

16. Shirley Navis M, Subila S. Study on the presence of oxalate ions in guava and sapota fruits. International Journal of Advanced Science and Research. 2017; 2(1): 15-17. Available from: http://www.allsciencejournal.com/archives/2017/vol2/issue1/1-12-17

17. Silgleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 1999; 299:152-178. Available from: doi.org/10.1016/S0076-6879(99)99017-1

Published

2021-09-14
Statistics
Abstract Display: 0
PDF Downloads: 0

Issue

Vol. 12 No. 3 (2021): Volume 12,Issue3

Section

Articles