Phytochemical composition of the underground part of Paeonia tenuifolia L. and leaves of Petrocoma hoefftiana (Fisch.) Rupr.

. The phytochemical composition of plants is very diverse and includes many biologically active substances, and their combination determines the pharmacological activity. This article examines the phytochemical composition of the underground part of Paeonia tenuifolia L. and leaves of Petrocoma hoefftiana (Fisch.) Rupr. It was found that the studied objects contain glycyrrhizic acid, peucedanin, flavonoids, phloroglucides, tannins, ascorbic acid. The studied plants can be considered as a new type of plant raw materials, promising for further study.


Introduction
This research was conducted within the scope of the scientific research direction of the ecological, chemical, and biological characteristics of rare and endangered plant species that occur on the territory of Kabardino-Balkaria and are suitable for use as therapeutic raw materials and decorative purposes.
The Paeonia tenuifolia L. is a rare species listed in the Red Book of the USSR (1984) and the Kabardino-Balkarian Republic (2000).
Popular names for it include funnel, lazarus flower, red lazorik, and voronets zelenika. It has disjunctive populations in the Pre-Caucasus, Central Caucasus, and Transcaucasia. Herbaceous polycarpic grows to a height of 20-50 cm, with tuberous thickenings and roots in rhizomes. It blooms in early April to early May. Seminal and vegetative reproduction are used. The slow development of seedlings and the lengthy germination of seeds are distinguishing characteristics. It has a strong regenerative ability: tiny rhizome segments can give rise to a new plant [1]. It occurs in Kabardino-Balkaria's grassland, where practically all age groups with a predominance of generative individuals are present in populations [2]. The Paeonia tenuifolia is an ornamental species that has been introduced into many botanical gardens. This species has been grown in the botanical garden of Kabardino-Balkarian State University. In medicine, preparations based on the Paeonia tenuifolia are used to treat heart disease, pulmonary tuberculosis, anemia, and various poisonings. They are bactericidal, protistocidal, and phytoncidal. Despite its widespread use in medicine, this plant's chemical composition has received little attention. The references contain information about the ascorbic acid content of Paeonia tenuifolia leaves, flavonoids and anthocyanins in flowers and pollen, and fatty oils in seeds [3].
Petrocoma hoefftiana (Fisch.) Rupr. is an endemic species of the Central Caucasus. The Red Book of the USSR (1984) and the Kabardino-Balkarian Republic (2000) both list it. It grows on limestone ridge cliffs, on rocks, and in narrow shaded gorges from the middle mountains to the Alpine belt. The populations are insignificant. Herbaceous is a perennial with creeping, strongly branched, brittle stems that grow to be 20-25 cm long. The leaves are ovate, pointed, and up to 3 cm long. The entire plant is pubescent, with white hairs protruding from it. The flowers are single or in groups of three, small, white, and occasionally pink. Slightly swollen Calyx srostnolistnaya. The petals in the upper part are divided into two parts, forming the number 5. The fruit is a single-nest, nearly spherical capsule that opens with denticles in the upper part when ripe. [1]. It is scientifically interesting because it is a paleo-endemic [4]. There is almost no information in the literature about the phytochemical composition of Petrocoma hoefftiana, however, the content of flavonoids in the ground part of the plant is known [5].
Plant species used in medicine contain biologically active substances that have a variety of therapeutic effects. These substances have a diverse composition and belong to various chemical compound classes. One of the most important tasks is to investigate the chemical composition of plants.

Materials and methods
During mass flowering, the leaves of generative individuals of Paeonia tenuifolia and Petrocoma hoefftiana were collected for analysis. To collect analytical samples for chemical analysis, GOST 24027.0-80 [6] was used. Saponins, coumarins, flavonoids, carotenoids, alkaloids, tannins, phloroglucides, cardiac glycosides, glycoalkaloids, and ascorbic acid were found in the underground parts of Paeonia tenuifolia and Petrocoma hoefftiana leaves.
An aqueous infusion (1:10) was used to determine saponins and ascorbic acid. 1 g of crushed vegetable raw materials was heated for 10 minutes in a water bath with 10 ml of distilled water. After cooling, the infusion was filtered.
Coumarins, flavonoids, and vitamin E were identified using alcohol extraction. In a 25 ml flask, 1 g of crushed raw materials was placed, and 10 ml of ethyl alcohol was added. The flask was connected to a reverse refrigerator and boiled in a water bath for 10 minutes. After cooling, the resulting extraction was filtered through a paper filter.
Tannin levels were determined using an aqueous extraction. 1 g of crushed vegetable raw materials was added to 100 ml of water. It was filtered through cotton wool after 30 minutes in a water bath.
Acetic acid extraction was used to determine glycoalkaloids. The crushed vegetable raw materials were mixed with a 5% acetic acid solution in a 1:10 ratio, shaken on a vibrator for 40 minutes, and then filtered through a paper filter.
Alkaloids were determined in hydrochloric acid extraction. 1 g of crushed vegetable raw materials was placed in a flask with a capacity of 100 ml, 25 ml of 1% HCl was poured and heated in a boiling water bath for 5 minutes. After cooling, the extraction was filtered through a paper filter.
In chloroform extraction, phloroglucides were determined. 1g of crushed vegetable raw materials were placed in a flask with a lapped stopper, filled with four times the amount of chloroform, and shaken periodically for a day. The mixture was then filtered through a paper filter.
Qualitative and quantitative determinations were carried out using well-known methods [7].

Results
Aqueous extracts (1:10) were prepared for the saponin presence study. Qualitative reactions (with lead acetate and the Lafon reaction) confirmed the presence of steroid saponins. Glycyrrhizic acid was discovered in the objects studied. The spectrophotometric method was used to determine the quantity of glycyrrhizic acid. It was discovered that its content is 500 mg% in the rhizomes of Paeonia tenuifolia and 9715 mg% in the leaves of Petrocoma hoefftiana.
To identify flavonoids, alcohol extracts (1:10) were prepared. A reaction with basic lead acetate and a cyanidine sample confirmed the presence of flavonoids in the analyzed objects. Rutin was discovered in both of the examined objects. The spectrophotometric method was used to determine the quantitative content of the total flavonoids. The rhizomes of Paeonia tenuifolia contain 90 mg% flavonoids, while the leaves of Petrocoma hoefftiana contain 140 mg%.
The content of hydrolyzable tannins in the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana has been qualitatively established.
The hydrolyzable tannin content of the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana has been quantified.
To identify phloroglucides, chloroform extracts were prepared (1:10). Their absence in the leaves of Petrocoma hoefftiana has been established, as has the presence in the rhizomes of Paeonia tenuifolia. The amount of phloroglucides in raw filicin was determined quantitatively using the pharmacopoeia method for male fern rhizomes. Filicin content was found to be 880 mg% in the rhizomes of Paeonia tenuifolia.
The qualitative reactions to the content of ascorbic acid in the aqueous infusion of the rhizomes of the Paeonia tenuifolia and the leaves of Petrocoma hoefftiana revealed its presence in the studied objects. The rhizomes of Paeonia tenuifolia contain 3300 mg% ascorbic acid, while the leaves of Petrocoma hoefftiana contain 44 mg%.
The absence of qualitative reactions to vitamin E group substances, carotenoids, alkaloids, glycoalkaloids, anthraglycosides, and cardiac glycosides was observed in both analyzed objects. Table 1 shows the results of a study of the chemical composition of the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana.

Discussion
Saponin-containing compounds have been discovered in the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana. Saponin-containing plants are used as expectorants, diuretics, and choleretics in medicine. The presence of saponins was discovered in aqueous infusions (1:10) of Paeonia tenuifolia rhizomes and Petrocoma hoefftiana leaves via qualitative reactions (reactions to foaming, with lead acetate, and the Lafon reaction). The precipitation of a significant amount of sediment in the reaction with lead acetate, as well as the appearance of intense blue-green staining in the Lafon reaction, lead us to believe that the raw material contains a significant amount of saponins. The results of the foaming reaction indicate the presence of steroid saponins. A specific reaction to the presence of glycyrrhizic acid, a saponin of the steroid group, yielded a positive result as well. We quantified glycyrrhizic acid in the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana using the spectrophotometric method. Petrocoma hoefftiana leaves outperform licorice roots as vegetable raw materials in terms of glycyrrhizic acid content [8].
Substances from the group of flavonoids with P-vitamin activity were discovered in the composition of the studied objects, which have a bactericidal, choleretic effect, and contribute to the removal of radioactive substances from the body.
Reactions were carried out with basic lead acetate and a cyanidine sample. The presence of flavonoids is confirmed by a qualitative reaction to rutin with 1% iron (III) chloride solution. The flavonoids in the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana were quantified using the spectrophotometric method. The flavonoid content of the studied objects is comparable to the flavonoid content of sea buckthorn fruits, which is a medicinal plant raw material [9].
Coumarins, natural compounds with anticoagulant properties, were discovered in the leaves of Petrocoma hoefftiana. The studied objects are comparable to the roots of the rare medicinal plant mustard Morison, which was previously used as a raw material in the production of the drug Peucedanin [10].
The studied objects contained hydrolyzable tannins, which are esters of saccharides and phenolic carboxylic acids. In acidic or enzymatic hydrolysis, hydrolyzable tannins are broken down into their simplest components. Tannins are not poisonous. Tannin-containing plants are used as bactericidal agents in the treatment of gastrointestinal diseases. Tannins' anti-inflammatory effect is based on their interaction with protein substances, which results in the formation of a protective film on the mucous membranes, which prevents the inflammatory process from progressing further. Furthermore, tannins are used in poisoning with alkaloids and heavy metal salts.
A qualitative analysis for chloroglucides of chloroform extracts of Paeonia tenuifolia rhizomes and Petrocoma hoefftiana leaves was performed using reactions with 1% solution of iron (III) chloride, potassium hexacyanoferrate (III), and concentrated nitric acid. The rhizomes of Paeonia tenuifolia contained a significant amount of phloroglucides. Petrocoma hoefftiana leaves do not contain phloroglucides. This group of substances chemical composition has received insufficient attention. These compounds have a variety of biological effects, including choleretic, antiviral, and anti-narcotic properties. The raw filicin content of the rhizomes of Paeonia tenuifolia was quantified using the pharmacopoeia method for rhizomes of Filicis maris. A sufficient amount of raw filicin has been found in the object under investigation.
Ascorbic acid, which plays an important role as a biologically active drug, is one of the important components of the phytochemical composition of the studied plants. Vitamin C helps enzymes that catalyze the hydroxylation of lysine and proline work properly. In medicine, ascorbic acid is used to treat and prevent vitamin deficiency and hypovitaminosis. Adults require 48 to 108 mg of vitamin C per day, depending on age, gender, and work intensity. Ascorbic acid is also used as an antioxidant in the manufacture of edible fats and fruit juices to prevent the formation of carcinogenic nitrosoamines and nitrites, which are added to these products in meat and sausage products.
The presence of vitamin C (ascorbic acid) was discovered in the aqueous infusion of the rhizomes of Paeonia tenuifolia and the leaves of Petrocoma hoefftiana by qualitative reactions using known methods [7]. The amount of 2,6-dichlorophenolindophenol, which went to the oxidation of vitamin C, was used to quantify ascorbic acid in aqueous extracts. The ascorbic acid content of Petrocoma hoefftiana leaves is insignificant. The amount of vitamin C in the rhizomes of Paeonia tenuifolia is comparable to that found in the fruits of some sea buckthorn varieties.
We determined the content of extractive substances in dried samples of Paeonia tenuifolia rhizomes and Petrocoma hoefftiana leaves using the well-known Soxlet apparatus and hexane extraction [7]. The extract of the dried rhizomes of Paeonia tenuifolia is a brown mass, and the leaves of the Petrocoma hoefftiana are dark green. The amount of extractive substances that can be extracted from the studied objects was determined after hexane extraction and solvent distillation.
Furthermore, qualitative analyses were performed on carotenoids, vitamin E group substances, alkaloids, anthraglycosides, and glycoalkaloids. The results of the analyses show that these substances are not present in the studied objects.

Conclusion
The chemical composition of Paeonia tenuifolia rhizomes and Petrocoma hoefftiana leaves has been studied. The following biologically active substances were found in the studied objects: glycyrrhizic acid, peucedanin, flavonoids, phloroglucides, tannins, and ascorbic acid. The findings allow us to recommend Paeonia tenuifolia and Petrocoma hoefftiana for introduction not only as ornamental plants, but also as a new type of medicinal plant raw materials worth further investigation.