Flowers marigold fermentation influence on the flavonoids content and polyphenoloxidase activity

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Abstract

Introduction. Fermentation is a biochemical process that changes the qualitative and quantitative composition of biologically active substances (BAS) and, accordingly, may change the pharmacological activity and toxicity of medicinal plants. Fermentation usually carried out for fresh medicinal plant raw materials (MPRM). However, to date, there is no information on the relationship between enzyme activity (in particular, polyphenol oxidase) and the BAS content. In this work, calendula flowers standardized for flavonoids were used as a model object.

Purpose of the study. To assess the effect of calendula flower fermentation on the flavonoid content and polyphenol oxidase activity.

Material and methods. The object of the study was calendula flowers (Calendulae flos, Calendula officinalis L., Asteraceae). The following fermentation options were studied: hot fermentation at 40 and 60°С; fermentation under natural conditions; cold fermentation, as well as thermal and ultrasonic pre-treatment. Immediately after the completion of fermentation, heat and ultrasound treatment, the polyphenol oxidase activity was determined in the processed fresh MPRM using the method of A.N. Boyarkin.

After 6 hours of fermentation of fresh MPRM at 40 and 60°С; after 1 day of fermentation under natural conditions; after cold fermentation and ultrasound treatment, heat treatment was carried out to stop the polyphenol oxidase activity. The flavonoid content was determined spectrophotometrically by the reaction with aluminum chloride.

Results. Hot fermentation of fresh calendula flowers at 40°С for 2–3 hours leads to a significant increase in the flavonoid content; hot fermentation at 60°С reduces the its content. The flavonoid content and polyphenol oxidase activity at 40°С are higher than at 60°С. A reliable relationship was revealed between the flavonoid content and polyphenol oxidase activity. Cold fermentation reduces the flavonoid content and polyphenol oxidase activity. Natural fermentation for 3 days increases the flavonoid content.

Ultrasonic treatment significantly increases the flavonoid content. Heat treatment does not significantly increase the its content. Thermal inactivation of the enzyme after hot, cold and natural fermentation and ultrasonic treatment reduces the flavonoid content.

Conclusion. Among all the studied methods of pre-treatment of fresh calendula flowers (hot, cold and natural fermentation, heat and ultrasonic treatment with and without thermal inactivation), the most significant increase in flavonoid content was achieved with ultrasonic treatment.

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About the authors

Raman Iharavich Lukashou

Educational Institution “Belarusian State Medical University”

Author for correspondence.
Email: r_lukashov@mail.ru
ORCID iD: 0000-0001-9547-5372

PhD, Associate Professor, Head of the Pharmaceutical Chemistry Department, Candidate of Pharmaceutical Sciences

Belarus, Dzerzhinsky Ave., 83, Minsk, 220083

Natalia Sergeevna Gurina

Educational Institution “Belarusian State Medical University”

Email: nsgur@mail.ru
ORCID iD: 0009-0009-9150-5728

Professor, Dean of the Pharmaceutical Faculty, Doctor of Biological Sciences

Belarus, Dzerzhinsky Ave., 83, Minsk, 220083

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2. Fig. 1. Dependence of flavonoid content and polyphenol oxidase activity on fermentation duration at 40°С

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3. Fig. 2. Dependence of flavonoid content and polyphenol oxidase activity on fermentation duration at 60°С

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4. Fig. 3. Flavonoid content and polyphenol oxidase activity of native and pre-treated medicinal plant raw material Note. 1. 1 – native medicinal raw plant material (MRPM); 2 – heat-treated MRPM; 3 – ultrasonic-treated MRPM; 4 – cold fermentation without packaging MRPM; 5 – cold fermentation in polyethylene packaging MRPM.

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5. Fig. 4. Dependence of flavonoid content and polyphenol oxidase activity on fermentation duration under natural conditions in polyethylene packaging and during natural drying

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6. Fig. 5. Flavonoid content in native and pre-treated medicinal plant raw material after thermal inactivation of enzymes Note. 1 – native medicinal plant raw material (MRPM); 2 – MRPM processed by ultrasound and then heat-treated; 3 – MRPM subjected to cold fermentation and then heat-treated; 4 – MRPM subjected to fermentation at 60°С and then heat-treated; 5 – MRPM subjected to fermentation at 40°С and then heat-treated; 6 – MRPM subjected to fermentation under natural conditions and then heat-treated.

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7. Fig. 6. Dependence of flavonoid content and polyphenol oxidase activity of dried calendula flowers on the duration of fermentation at 40°С

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8. Fig. 7. Effect of cold fermentation on the yield of flavonoids from dried calendula flowers

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