Analysis of the Composition and Quantitative determination of Hydroxycinnamic acids in Quince fruits cultivated in the Russian Federation

Nesterova N. V.¹, Teplov I. S.², Bokov D. O.², Sergunova E. V.², Luferov A. N.²,

Samylina I. A.², Averseva I. N²

¹Peoples ' Friendship University, of Russia 117198, Moscow Miklukho-Maklaya str.6.

²Sechenov First Moscow State Medical University, 8 Trubetskaya St.,

bldg. 2, Moscow, 119991, Russian Federation.

*Corresponding Author E-mail: nestero-nadezhda@yandex.ru

ABSTRACT:

Objective: Considering the growing interest of researchers in a wide range of pharmacological effects characteristic of quince raw materials due to the presence of various groups of biologically active substances (BAS). The object of the study was fruits Cydonia oblonga Mill obtained in the Botanical Garden of Sechenov University, the Botanical Garden of Moscow State University, the Moscow and Krasnodar regions, harvested during the fruiting phase. Materials and methods: The analysis was performed on a GILSON high-performance liquid chromatograph, model 305 (France). Detection was carried out using a GILSON UV/VIS model 151 UV detector at a wavelength of 370 nm. The total content of polyphenols in the studied raw materials was estimated by Folin-Chicalteu spectrophotometry, the quantitative content of hydroxycinnamic acids in quince fruits was determined by spectrophotometry in terms of caffeic acid Results and discussions: The authors studied the composition of polyphenolic compounds and carried out their quantitative assessment, using HPLC analysis, the presence of polyphenolic substances represented by caffeic, gallic, chlorogenic, neochlorogenic, cinnamic acids, as well as catechin, quercetin, floretin and apigenin was established. A quantitative assessment of the total content of polyphenolic substances in fruits was carried out using the Folin-Chicalteu method, during which it was revealed that the content of the polyphenolic complex ranges from 8,775 to 9,221 in fresh raw materials and from 6,799 to 7,676 in dried raw materials. Losses of phenolic compounds during drying range from 16.8 to 22.5%. The total content of hydroxycinnamic acids in terms of caffeic acid was carried out by spectrophotometric analysis. The results obtained showed the content of analyzed substances in the determined raw materials from 2,021 to 2,344 in dried raw materials and from 3,126 to 3,526 in fresh. Conclusion: The analysis makes it possible to identify phenolic derivatives in extracts from quince fruits, which simplifies the standardization of raw materials in the preparation of appropriate regulatory documentation

KEYWORDS: Common quince, Quince fruit, Polyphenolic Compounds, Hydroxycinnamic acids, HPLC, Spectrophotometry.

INTRODUCTION:

The Caucasus and Transcaucasia are considered to be the birthplace of quince, from where it migrated to Asia Minor and then to the ancient Greeks. According to the literature, the first mention of common quince (Cydonia oblonga Mill) was found in ancient Greek manuscripts dating back to the 1st century BC. e. The ancient Greek physician Hippocrates recommended quince fruits to improve the gastrointestinal tract. Pliny the Elder, an ancient Roman philosopher and scientist, described several varieties of quince in his scientific treatises, drawing attention to the possibility of using sweet varieties for fresh food, and more tart ones for baking and use for medicinal purposes. In those days, a decoction of sour-tart wild quince fruit was used by healers to improve bowel function, jaundice, diarrhea, and palpitations.

Even in the Middle Ages, it was noticed that quince promotes digestion, so they stewed quince together with meat. Thus, they pursued another goal, improving the quality of the dish, because this fruit is very fragrant. If the quince is cooked, baked or stewed for a long time, its flesh becomes a beautiful rich red color. Since it is rich in pectin substances, it is used for making jams and jellies. In the literature, there are interesting data that the first marmalade was made in ancient times from quince.

In India, the fruit is used for stomach and intestinal diseases, as it improves digestion. The best remedy for dysentery is considered to be unripe fruits, especially in young children. Mature fruits have the properties of a gentle laxative. Unripe fruits have a milky effect. The fruit is widely used both as a dessert fruit, and for the preparation of aromatic and refreshing soft drinks.

In Tibetan medicine, quince was used for “hot diarrhea”, “Bad-kan” diseases associated with metabolic disorders in the body (“Bad-kan” in Tibetan medicine is a regulator of fat and water metabolism). In modern therapeutic practice, a decoction of quince fruit is widely used in eastern medicine, noting the presence of antiemetic, anti-rheumatic, antispasmodic, anti-inflammatory, analgesic, astringent effects.

The wide experience of traditional medicine and the recommendations of a number of phytotherapists on the use of representatives of the Rosaceae family, including quinces for the prevention and treatment of various diseases, determine the constant growing interest of researchers both in the pharmacological action of this raw material and in a comprehensive study of the chemical composition. Scientists have established the thrombolytic activity of Prunus avium extract¹, the antimicrobial activity of Pyrus communis against Staphylococcus aureus, Bacillus cereus, Staphylococcus subtilis and Escherichia coli², and the anti-inflammatory effect of Rubus ulmifolius³and the adsorption capacity of Prunus cerasus⁴. The relationship between the phenolic fraction and the antimicrobial activity of Prunus domestica has been revealed.⁵ The relationship between the phenolic fraction and the antimicrobial activity of P. domestica has been revealed. The scientific review provides data on the phytochemical composition of Prunus amygdalus and the possibilities of its pharmacological use.⁶ The composition of biologically active substances Rubus racemosus⁷ and Malus domestica i.e Gala and fuji apple has been revealed.⁸ An interesting area of research is the use of representatives of the Rosaceae family in functional nutrition products in combination with other plant objects.⁹ The analysis of the phytochemical properties of the Rosaceae family uses techniques used for other plant objects.¹⁰

According to the literature data, ethanol extracts of quince fruits have an antimicrobial effect, inhibiting the growth of Escherichia coli, and Staphylococcus aureusbacteria, as well as an antifungal effect against Aspergillus niger.¹¹ Cardioprotective action was revealed, which is associated with the presence of kaempferol-3-O-glycoside (astragalin) in raw materials. It was experimentally established that the use of captopril at a dosage of 25mg/kg led to a decrease in blood pressure in rats after two weeks, while ethanol extraction of quince fruits and leaves reduced blood pressure after four weeks at doses of 80 and 160mg/kg of body weight. In addition, water quince extracts improved thrombolysis compared to aspirin. Quince has been shown to have the potential to prevent thrombosis and reduce the risk of cardiovascular diseases. A number of scientific studies are devoted to the study of the antioxidant effect of extracts from quince fruits, and the presence of an antioxidant effect exceeding the effect of a number of previously studied fruits has been experimentally established.

It should be noted that most researchers associate the therapeutic activity of quince fruits with the polyphenol complex contained in them Considering the above, the aim of our work is to compare the composition and quantitative content of polyphenolic compounds of fresh and dried quince fruits.

MATERIAL AND METHODS:

The object of our research was quince fruits harvested from cultivated plants growing in the Botanical Gardens of Sechenov University, Lomonosov Moscow State University, harvested during the fruiting phase (September 2023).

All reagents used in the study had a degree of purity of ch. d. a. The qualitative composition of biologically active substances was studied in extracts from quince fruits obtained by extraction with 70% ethyl alcohol, using a high-performance liquid chromatograph from GILSON, model 305 (France), manual injector, model RHEODYNE 7125 USA, followed by computer processing of the study results using the Multichrom program for Windows. A metal column of 4.6 x 250mm KROMASIL C18 with a particle size of 5 microns was used as a stationary phase tbe. A mixture of acetonitrile – water – phosphoricacid, concentrated in the ratio 20: 80: 0.05 was used as the mobile phase. The analysis was performed at room temperature. The eluent feed rate is 1.0ml/min. The analysis duration is from 40 to 60 minutes. Detection was performed using a “GILSON” UV/ VIS model 151 UV detector at a wavelength of 370 nm.

The total content of polyphenols in the studied raw materials was estimated by the Folin-Chicalteu spectrophotometry method, which is widely used in the analysis of plant raw materials for medicinal and food purposes.¹²

Determination of the total content of hydroxycinnamic acids (HAs) in quince fruits was carried out by spectrophotometry in terms of caffeic acid (3,4-dioxycinnamic acid, 3 - (3,4-dihydrophenyl) - 2-propenic acid), which is due to the immunomodulating, antibacterial, antimycotic and antioxidant activity proven for this compound.^13-23 Pre-ground to the size of particles, passing through a sieve with a hole diameter of 2mm, raw materials weighing 2g (exact weight) were placed in a flask with a capacity of 200ml and 70ml of distilled water was added. The flask was attached to a return refrigerator and heated in a water bath for 15 minutes. Extraction was repeated. The resulting extracts were cooled at room temperature and filtered through a paper filter. The extracts were quantitatively transferred to a 200ml volumetric flask and the volume of the distilled water solution was adjusted to the mark. Then, 1ml of the previously obtained quince fruit extract was transferred to a 50ml volumetric flask and the volume of the solution was adjusted to sign using 20% ethyl alcohol. The optical density of the resulting solution was measured at a wavelength of 325nm, since this value is analytical for caffeic acid. 20% ethyl alcohol was used as the reference solution.

The content of the sum of HAs (X, %) in the leaves of a forest apple tree in terms of coffee acid was calculated by the formula:

Ax x 200 x 50

X= ---------------------------------------------------- x 100

E^1%_1cm x m x V_vax (100-W)

Where Ax is the optical density of the test solution; E^1%_1cm is the specific absorption index of caffeic acid at 325 nm, which is 782; m is the weight of the suspension of crushed quince fruits; V_va– aliquot volume, ml; W – humidity, %.

Statistical processing of the results of the study was carried out in accordance with the requirements of GPM 42-0111-09 “Statistical processing of the results of a chemical experiment”. Validation of the method of quantitative assessment of HAs in the leaves of the forest apple tree was carried out in accordance with the recommendations for the following indicators: correctness, precision, specificity, linearity, analytical area (range of application).

RESULTS AND DISCUSSION:

Chromatographic analysis of ethanol extraction from quince fruits harvested during the fruiting phase showed the identity of biologically active substances of all the studied samples, the composition of which is presented in Table 1 and Figure 1.

Figure 1: Chromatogram of alcohol extraction from quince fruits by HPLC

Table 1: Results of the study of phenolic compounds in dry and fresh raw materials-quince fruits, by HPLC.

No	Time, min	Formula	Compound name
1	4.456		Caffeicacid(3,4-dioxicinnamic acid)
2	5.214		Gallic acid (3,4,5-trihydroxybenzoic acid)
3	6.161		Catechin ((2R, 3S) - 2- (3,4-dihydroxyphenyl) - 3,4-dihydro-2H-chromene-3,5,7-triol)
4	7.904		Chlorogenic acid ((1S, 3R, 4R, 5R) -3-{[(2E) - 3- (3,4-dihydroxyphenyl) prop-2-enoyl] oxy} - 1,4,5 trihydroxycyclohexane-1-carboxylic acid)
5	9.957		Non chlorogenic acid ((1R, 3R, 4S, 5R) -3 - {[(2E) - 3 - (3,4-dihydroxyphenyl) prop-2-enoyl] oxy} - 1,4,5-trihydroxycyclohexane-1-carboxylic acid)
6	18, 523		Phloretin (3-(4-hydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)propane-1-one)
7	20.355		Quercetin (2-(3,4-dihydroxyphenyl) - 3,5,7-trihydroxy-4H-chromene-4-one)
8	21.113		Cinnamiс acid (E) - 3-phenylpropenic acid)
9	22.959		Apigenin (4H-1-Benzopyran-4-one, 5,7-dihydroxy-2 - (4-hydroxyphenyl) - 5,7-Dihydroxy-2-(4-hydroxyphenyl) - 4H-1-benzopyran-4-one)

Table 3. Results of quantitative determination of hydroxycinnamic acids in quince fruits in terms of coffee acid in fresh and dried raw materials

Sample No	Test object	Content of hydroxycinnamic acids in terms of caffeic acid %	Content of hydroxycinnamic acids in terms of caffeic acid in fresh raw materials, %
1	Quince fruit (Sechenov University Botanical Garden)	2.09±0.89	3.13±0.93
2	Quince fruit (MSU Botanical Garden)	2.14±0.85	3.37±0.89
3	Quince fruit (Moscow region)	2.02±0.94	3.41±0.91
4	Quince fruit (Krasnodar Region)	2.34±0.70	3.53±0.78


Figure 2: Results of quantitative determination of the total content of polyphenolic compounds in quince fruits in terms of fresh and dried raw materials by the Folin-Chicalteu method	Figure 3: Results of quantitative determination of hydroxycinnamic acids in quince fruits in terms of coffee acid in fresh and dried raw materials

As can be seen from the table, the content of the polyphenol complex ranges from 8.775 to 9.221 in fresh raw materials and from 6.799 to 7.676 in dried raw materials. Losses of phenolic compounds during drying range from 16.8 to 22.5%

The results of quantitative determination showed the content of analytes in the determined raw materials from 2.021 to 2.344 in dried raw materials and from 3.126 to 3.526 in fresh ones.

CONCLUSIONS:

The results of the study were a detailed analysis of the composition of polyphenolic compounds and their quantitative assessment. The authors used the method of HPLC analysis, and found the presence of polyphenolic substances represented by caffeic, gallic, chlorogenic, non-chlorogenic, cinnamic acids, as well as catechin, quercetin, phloretin and apigenin.

The total content of polyphenolic substances in fruits was quantified by the Folin-Chicalteu method. The authors found that the content of the polyphenol complex ranges from 8.775 to 9.221 in fresh raw materials and from 6.799 to 7.676 in dried raw materials. Losses of phenolic compounds during drying range from 16.8 to 22.5%.

Quantitative assessment of the total content of HAs in terms of caffeic acid was carried out by spectrophotometric analysis. The results obtained showed the content of analytes in the determined raw materials from 2.021 to 2.344 in dried raw materials and from 3.126 to 3.526 in fresh ones.

CONFLICTS OF INTEREST:

None.

ACKNOWLEDGEMENTS:

This research has been supported by the RUDN University Strategic Academic Leadership program grant number 033323-2-000.

References:

1. Patel V. Manek RA. Sheth DB. Evaluation of in-vitro Thrombolytic activity of methanolic extract of Prunus avium L. Asian Journal of Research in Pharmaceutical Sciences. 2021; 11(1): 41-4, doi: 10.5958/2231-5659.2021.00007.2

2. Kiran Garg V. Dhiman A. Evaluation of Antimicrobial Activity of Peel and Fruits of Pyrus communis. Research Journal of Pharmacy and Technology. 2020; 13(1): 293-6, doi: 10.5958/0974-360X.2020.00059.1

3. Pandey R. Pandey R. Shukla SS. Anti-inflammatory potential of ethanol extract of Rubus ulmifolius (Schott). Research Journal of Pharmacy and Technology. 2013; 6(3):300-3.

4. Kale AA. Kinetic and Thermodynamic Study of Adsorption Methylene Blue by Nitrated Biomass of Prunus Cerasus. Asian Journal of Research in Chemistry. 2021; 14(4): 242-6, doi: 10.52711/0974-4150.2021.00041

5. Upyr TV. Lenchyk LV. Komisarenko MA. Navruzzoda GF. Komisarenko AM. Osolodchenko TP. Ponomarenko SV. Study of Phenolic Compounds of Prunus Domestica Presscake and Antimicrobial Activity of Its Extract. Research Journal of Pharmacy and Technology. 2022; 15(1): 375-80, doi: 10.52711/0974-360X.2022.00061

6. Singh D. Gohil KJ. Rajput RT. Sharma V. Almond (Prunus amygdalus Batsch.): A Latest Review on Pharmacology and Medicinal uses. Research Journal of Pharmacy and Technology. 2022; 15(7): 3301-8, doi: 10.52711/0974-360X.2022.00553

7. Kumar PR. Vaidhyalingam V. Pharmacognostical and Preliminary Phytochemical Studies on the Aerial Parts of Rubus racemosus (Roxb). Research Journal of Pharmacognosy and Phytochemistry. 2010; 2(5):381-5.

8. Ahmad S. Mahmood T. Kumar R. Bagga P. Ahsan F. Shamim A. Ansari A. et al A Contrastive Phytopharmacological Analysis of Gala and Fuji Apple. Research Journal of Pharmacy and Technology. 2020; 13(3): 1527-37, doi: 10.5958/0974-360X.2020.00278.4

9. Thiruvengadam S. Naresh B. Nivedhaa GK. Ivoromauld S. Preparation of Fruit Leather and Fortification with Moringa oleifera. Research Journal of Pharmacy and Technology. 2020; 13(4): 1619-22, doi: 10.5958/0974-360X.2020.00293.0.

10. Raj A. Vinnarasi J. Venkataraman R. Augustin M. HPTLC Fingerprinting Analysis of Tannin Profile on Canthium coromandelicum and Flueggea leucopyrus willd. Research Journal of Pharmacy and Technology. 2018; 11(12): 5355-8. doi: 10.5958/0974-360X.2018.00975.7

11. Herrera-Rocha KM. Rocha-Guzmán NE. Gallegos-Infante JA. González-Laredo RF. Larrosa-Pérez M. Moreno-Jiménez MR. Phenolic Acids and Flavonoids in Acetonic Extract from Quince (Cydonia oblonga Mill.): Nutraceuticals with Antioxidant and Anti-Inflammatory Potential. Molecules. 2022; 27(8): 2462, doi: 10.3390/molecules27082462

12. Nikolaeva TN. Lapshin PV. Zagoskina NV. Method for Determining the Total Content of Phenolic Compounds in Plant Extracts with Folin–Denis Reagent and Folin–Ciocalteu Reagent: Modification and Comparison. Chemistry of plant raw material. 2021; (2):291-9, doi: 10.14258/jcprm.2021028250

13. Ňorbová M. Vollmannová A. Fedorková S. Musilová J. Lidiková J. The forgotten fruit (Cydonia oblonga Mill.) and its chemical composition: a review. European Food Research and Technology. 2024; 250:2093–102, doi: 10.1007/s00217-024-04543-7

14. Dalkılıç LK. Investigation of Antioxidant, Antimicrobial and Cytotoxic Activity of Cydonia oblonga Leaf on Breast Cancer (MCF-7) and Liver Cancer (HepG2) Cell Lines. Turkish Journal of Science and Technology. 2024; 19(1): 13-23, doi: 10.55525/tjst.1331063

15. Al-Zughbi I. Krayem M. Quince fruit Cydonia oblonga Mill nutritional composition, antioxidative properties, health benefits and consumers preferences towards some industrial quince products: A review. Food Chemistry. 2022; 393: 133362, doi: 10.1016/j.foodchem.2022.133362

16. Lukashov RI. Moiseev DV. Stolyarova VN. Makarenko MN. Pharmacological activity of caffeic acid. Bulletin of Pharmacy. 2012; 57(3): 61-5.

17. Abed SN. Bibi S. Jan M. Talha M. Islam NU. Zahoor M. Al-Joufi FA. Phytochemical Composition, Antibacterial, Antioxidant and Antidiabetic Potentials of Cydonia oblonga Bark. Molecules. 2022; 27(19):6360, doi: 10.3390/molecules27196360

18. Najman K. Adrian S. Sadowska A. Świąder K. Hallmann E. Buczak K. Waszkiewicz-Robak B. Szterk A. Changes in Physicochemical and Bioactive Properties of Quince (Cydonia oblonga Mill.) and Its Products. Molecules. 2023; 28(7): 3066. doi: 10.3390/molecules28073066

19. Ivashev M. N., Chuklin R. E. Effect of oxycinnamic acids on the cerebral circulatory system. Pharmacy and pharmacology. 2013; 1(1): 44-48. doi: 10.19163/2307-9266-2013-1-1-44-48

20. Altuntas S. Korukluoglu M. Biological activity of optimized phenolic extracts of quince (Cydonia oblonga Miller) parts before and after simulated in vitro gastrointestinal digestion. Food Chemistry. 2024; 437: 137846, doi: 10.1016/j.foodchem.2023.137846

21. Golafshani MG. Tavakoli H. Hosseini SA. Akbari M. MD and DFT computational simulations of Caffeoylquinic derivatives as a bio-corrosion inhibitor from quince extract with experimental investigation of corrosion protection on mild steel in 1M H2SO4. Journal of Molecular Structure. 2023; 1275: 134701, doi: 10.1016/j.molstruc.2022.134701

22. Urbanavičiūtė I. Liaudanskas M. Bobinas Č. Šarkinas A. Rezgienė A. Viskelis P. Japanese Quince (Chaenomeles japonica) as a Potential Source of Phenols: Optimization of the Extraction Parameters and Assessment of Antiradical and Antimicrobial Activities. Foods. 2020; 9(8):1132, doi: 10.3390/foods9081132

23. Rather JA. Yousuf S. Ashraf QS. Mir SA. Makroo HA. Majid D. Barba FJ. Dar BN. Nutritional and bioactive composition, nutraceutical potential, food and packaging applications of Cydonia oblonga and its byproducts: A review. Journal of Food Composition and Analysis. 2023; 115: 105000, doi: 10.1016/j.jfca.2022.105000

24. Nesterova NV. Matveenko VN. Samylina IA. Kondrashev SV. Bobkova NV. Suleymanova FS. The Detection of Floretin and Floridzin in Eastern Apple Tree Fruits (Malus orientalis Uglitzk. ex Juz.) and Assessment of the Quantitative Content of Phenolic Compounds. Moscow University Chemistry Bulletin. 2021; 76(1): 71-4. doi: 10.3103/S0027131421010107

Received on 25.09.2024 Revised on 15.01.2025

Accepted on 30.03.2025 Published on 02.05.2025

Available online from May 07, 2025

Research J. Pharmacy and Technology. 2025;18(5):2291-2296.

DOI: 10.52711/0974-360X.2025.00328

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Sample No	Sample Name	Polyphenol content in fresh raw materials, %	Polyphenol content in dried raw materials, %
1	Quince fruit (Sechenov University Botanical Garden)	8.93±0.96	6.98±0.85
2	Quince fruit (MSU Botanical Garden)	8.77±0.74	6.79±0.92
3	Quince fruit (Moscow region)	8.82±0.92	6.91±0.88
4	Quince fruit (Krasnodar Region)	9.22±0.72	7.67±0.75