Rosa davurica Pall., Rosa rugosa Thumb., and Rosa acicularis Lindl. Originating from Far Eastern Russia: Screening of 146 Chemical Constituents in Three Species of the Genus Rosa

: Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. contain a large number of target analytes which are bioactive compounds. High performance liquid chromatography (HPLC), in combination with the ion trap (tandem mass spectrometry), was used to identify target analytes in MeOH extracts of R. rugosa , R. davurica , and R. acicularis , originating from the Russian Far East, Trans-Baikal Region, and Western Siberia. The results of initial studies revealed the presence of 146 compounds, of which 115 were identiﬁed for the ﬁrst time in the genus Rosa (family Rosaceae ). The newly identiﬁed metabolites belonged to 18 classes, including 14 phenolic acids and their conjugates, 18 ﬂavones, 7 ﬂavonols, 2 ﬂavan-3-ols, 2 ﬂavanones, 3 stilbenes, 2 coumarins, 2 lignans, 9 anthocyanins, 3 tannins, 8 terpenoids, 3 sceletium alkaloids, 4 fatty acids, 2 sterols, 2 carotenoids, 3 oxylipins, 3 amino acids, 5 carboxylic acids, etc. The proven richness of the bioactive components of targeted extracts of R. rugosa , R. davurica , and R. acicularis invites extensive biotechnological and pharmaceutical research, which can make a signiﬁcant contribution both in the ﬁeld of functional and enriched nutrition, and in the ﬁeld of cosmetology and pharmacy. lignans: Pinoresinol, Arctigenin; coumarins: 3,4,5–Trimethoxycoumarin, Fraxin; anthocyanins: Cyanidin 3- O -glucoside, Delphinidin O -pentoside, Pelargonidin 3- O -(6- O -malonyl-β - D -glucoside), Cyanidin (cid:48)(cid:48) -Succinyl-Glucoside), Cyanidin O -dioxayl-glucoside,


Introduction
Plants have been used as medicines since the existence of human civilization [1,2]. More than 35 thousand varieties of plants from different parts of the world are actively used for medical purposes, since they contain numerous phytocomponents that can potentially treat many diseases, including infectious ones [3]. Numerous medical systems of treatment, such as Ayurveda, Unani, homeopathy, naturopathy, Siddha, and others, rely on plants as effective remedies for various life-threatening diseases [4,5]. Due to the presence of secondary metabolites in plants, they have significant potential as antimicrobial agents. The diversity of these natural products offers an endless number of possibilities for the discovery of new drugs for the treatment of various diseases [6][7][8].
presence of secondary metabolites in plants, they have significant potential as antimicrobial agents. The diversity of these natural products offers an endless number of possibilities for the discovery of new drugs for the treatment of various diseases [6][7][8].
In recent years, traditional medicine based on oral herbal preparations has attracted the attention of both consumers and healthcare professionals. However, the use of these medicinal products requires improved knowledge of their composition and stability over time in order to support or validate these therapies in humans. Liquid preparations from medicinal plants, such as tinctures and extracts from plant buds, are typical products that are widely used but still poorly understood. Plant bud extracts are defined as extracts obtained exclusively from fresh buds, shoots, young leaves, and/or roots, which are macerated and extracted with hydro-glycerol and water-alcohol mixtures [9]. Kidney extracts represent a new category of herbal products well known and widely used in gemmotherapy, as well as in homeopathy and herbal medicine [10].
The genus Rosa (family Rosaceae) is represented on the territory of the Trans-Baikal region, Far East (Russian Federation), and Western Siberia by 3 species-Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. (Figures 1 and 2). Fresh fruits and leaves contain up to 900 mg% ascorbic acid per dry pulp weight. Fresh petals contain 0.25-0.38% essential oil. Its neutral volatile fraction contains 86.3% phenylethyl alcohol, some linalool, citronellol, geraniol, nerol, etc. Eugenol was found in the phenolic fraction, phenylacetic, benzoic, and other acids in the acid fraction. R. rugosa is a medicinal plant widely used in traditional and folk medicine. Extracts of R. rugosa have been valued for Asian culinary, cosmetic, and aromatherapy purposes, and used in herbal medicines for diabetes mellitus and osteoarthritis. [11]. The medicinal effects seem to be involved in the presence of many phytochemicals in R. rugosa extracts, for example flavonoids, phenylpropanoid, tannins, fatty acids, and terpenoids [12].  Several studies have reported that some compounds from rose hip extracts exhibit anti-inflammatory activity in vitro. The anti-inflammatory property of the crude hydroalcoholic extract of rose hip has been proven in vivo, suggesting its potential role as one of Several studies have reported that some compounds from rose hip extracts exhibit antiinflammatory activity in vitro. The anti-inflammatory property of the crude hydroalcoholic extract of rose hip has been proven in vivo, suggesting its potential role as one of the main therapies for the treatment of diseases associated with inflammation [13]. In Turkish folk medicine, a decoction of fresh rose hips is prepared and used to treat various stomach disorders [14]. Trans-Tiliroside (Tribuloside) has been found to be one of the main active components of aqueous acetone extracts from fruits and seeds that inhibit weight gain and lower plasma triglyceride levels in mice [15]. Additionally, clinical studies have demonstrated the positive effect of rose hip powder in the treatment of osteoarthritis [16]. Rose hip powder enhances in vitro anti-inflammatory and chondroprotective properties in leukocytes and primary chondrocytes of human peripheral blood [17]. Unfortunately, to date, there are few data providing information on the biological action of extracts of buds and leaves, and it should be noted that these preparations have never been used for preclinical and clinical trials.
The present investigation was designed to carry out a phytochemical study involving detailed metabolomic and comparative analysis of fruits and flowers of Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. originating from the Trans-Baikal region, Western Siberia, and Russian Far East.

Results
Approximately 300 mass spectra were assessed for each analytical replicate and MS operating condition in this comprehensive approach for a complete screening of phytochemicals ( Figure 3). Several studies have reported that some compounds from rose hip extracts exhibit anti-inflammatory activity in vitro. The anti-inflammatory property of the crude hydroalcoholic extract of rose hip has been proven in vivo, suggesting its potential role as one of the main therapies for the treatment of diseases associated with inflammation [13]. In Turkish folk medicine, a decoction of fresh rose hips is prepared and used to treat various stomach disorders [14]. Trans-Tiliroside (Tribuloside) has been found to be one of the main active components of aqueous acetone extracts from fruits and seeds that inhibit weight gain and lower plasma triglyceride levels in mice [15]. Additionally, clinical studies have demonstrated the positive effect of rose hip powder in the treatment of osteoarthritis [16]. Rose hip powder enhances in vitro anti-inflammatory and chondroprotective properties in leukocytes and primary chondrocytes of human peripheral blood [17]. Unfortunately, to date, there are few data providing information on the biological action of extracts of buds and leaves, and it should be noted that these preparations have never been used for preclinical and clinical trials.
The present investigation was designed to carry out a phytochemical study involving detailed metabolomic and comparative analysis of fruits and flowers of Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. originating from the Trans-Baikal region, Western Siberia, and Russian Far East.

Trimethoxyflavones
The flavones Nevadensin (compound 12) and Pentahydroxy trimethoxy flavone (compound 15) ( found in extracts of R. acicularis. The CID-spectrum in positive ion modes of Nevadensin from extracts of R. acicularis is shown in Figure 5.
The fragment ion with m/z 298 yielded two daughter ions at m/z 271 and m/z 227. It was identified in the bibliography in extracts of Triticum aestivum [22]; millet grains [23]; Sasa veitchii; Phyllostachys nigra [24].

Condensed Tannin
Prodelphinidin A-type (compound 83) and (S)-Flavogallonic acid (compound 84 have been already characterized as components of Vitis vinifera [45], Terminalia arjuna [46] and R. rugosa [47]. CID-spectrum in positive ion modes of (S)-Flavogallonic acid from R davurica is shown in Figure 10 [46] and R. rugosa [47].   The polyphenol composition distribution table of varieties Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. is shown below [ Table 1]. The comparison table shows the presence of some polyphenols in three types of the genus Rosa (kaempferol, ellagic acid). Some polyphenols are present in only one variety of the genus Rosa.

Materials
Aboveground phyto Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. collected during expedition work on the territory of the Russian Far East, Trans-Baikal Region, and Western Siberia during the period of ripening (July-September, 2020). Phyto mass of R. davurica was collected on the territory of Buryatia (N 52 • 21 97 E 108 • 59 84 ), in September 2020. Phyto mass of R. rugosa was collected on the territory of Primorsky Krai, Russia (N 42 • 36 10 E 131 • 10 55 ), during the period from 10 to 20 August, 2020. Phyto mass of R. acicularis was collected on the territory of Kemerovo, Western Siberia (N 55 • 21 15 E 86 • 05 23 ), in August 2020. All samples were morphologically authenticated according to the current standard of Pharmacopoeia of the Eurasian Economic Union [48].
The results were obtained using the equipment of the Center for Collective Use of Scientific Equipment of TSU named after G.R. Derzhavin.

Chemicals and Reagents
HPLC-grade acetonitrile was purchased from Fisher Scientific (Southborough, UK), MS-grade formic acid was from Sigma-Aldrich (Steinheim, Germany). Ultra-pure water was prepared from a SIEMENS ULTRA clear (SIEMENS water technologies, Germany), and all other chemicals were analytical grade.

Fractional Maceration
To obtain highly concentrated extracts, fractional maceration was applied. In this case, the total amount of the extractant (methyl alcohol of reagent grade) is divided into 3 parts and is consistently infused on potato with the first part, then with the second and third. The infusion time of each part of the extractant was 7 days.

Conclusions
The extracts of Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. contain a large number of polyphenolic complexes which are biologically active compounds. For the most complete and safe extraction, the method of maceration with MeOH was used. To identify target analytes in extracts, HPLC was used in combination with the ion trap. The results of a preliminary study showed the presence of 146 bioactive compounds, of which 115 were identified for the first time in the genus Rosa (family Rosaceae). Of these 115 chemical compounds identified for the first time in the genus Rosa, 70 compounds belonged to the polyphenolic group: 18 flavones, 7 flavonols, 3 flavan-3-ols, 2 flavanones, 14 phenolic acids, 3 stilbenes, 2 lignans, 2 coumarins, 9 anthocyanidins, 3 tannins, etc. The proven richness of the bioactive components of targeted extracts of R. rugosa, R. davurica, and R. acicularis invites extensive biotechnological and pharmaceutical research, which can make a significant contribution both in the field of functional and enriched nutrition, and in the field of cosmetology and pharmacy. It should also be noted that the variability of the genus Rosa (family Rosaceae) contributes to the selection of the most drought-resistant species and samples for household, decorative, and forest reclamation needs in the arid climatic zones of Eurasia.
It is important to note that the useful properties of the genus Rosa (family Rosaceae) are: food (R. rugosa, R. acicularis), perfumery (R. acicularis, R. ecae), nectariferous (R. canina, R. cinnamomea), decorative (R. acicularis, R. rugosa), and soil-strengthening (R. acicularis, R. rugosa, R. spinosissima). A wide variety of biologically active polyphenolic compounds opens up rich opportunities for the creation of new drugs, as well as bioactive additives based on extracts from the genus Rosa.          [112,115] * Compounds identified for the first time in genus Rosa.