Among known the biologically active substances of plant, α-, β- and γ-pirons are very important and have physiological activities in various directions. They are the basis for the different drugs, which using the practice medicine as spasmolytic, anticoagulant, expanding capillaries, hypotensive, anticarcinogen, anti-inflammatory, bile repellent and etc. From ammifurin, psoralen, beroxanum, meladinine, methoxsalen, visnadin, dihydrosamidin, atamantin and pterixin that is widely used in medicine for treatment leucoderma, vitiligo. Various derivatives of 4-Oxycoumarin are the basis for the drugs dicoumarol and pelentan having anticoagulant effects. The substance 4.7-dioxy-3-amino-8- metylcoumarin obtained from 4.7-dioxy-3-amino-8-metylcoumarin Sreptomyces spheroids is highly active against the microorganisms and is a base for the antibiotic drug “novobiosin” [1,2].

The mix of extractive substances was obtained by finely cutting the body of Johrenia paucijuga, then drying in the room conditions (300 g) and extracting by acetone 3 times (3 days for each time). To obtain the extractive substances the mix was chromagraphed by the chromatography method in the glass column filled by Al₂O₃. The obtained substances were compared based on the ¹H and ¹³C NMR information provided in literature on the structure of the individual items.

It was identified 6350 higher plants, including 2500 regional endemic species in Caucasian flora [3]. About 900 medicinally important plants, 850 species with essential oil, 200 plants with rubber, 200 with vaccinations ingredients, 460 colored, 600 with alhoids, more than 400 plants with vitamins are known in Azerbaijan flora [4].

Medicinal plants are very important for human health and its strengthening. The drugs produced from these plants do not lead to dependence. More than 50% of the drugs used in the world are of vegetable origin. Johrenia paucijuga species belongs to the genus Umbelliferae. About 400 species and 3500 families are known in the temperate and subtropical regions of the Northern Hemisphere of the Earth [5].

The family of Umbelliferae are noticeably plants. They play a major role in the formation of vegetation and give the unique charms to the landscapes. Representatives of this family are perennial, biennial and annual herbs, rarely shrubs, bushes and small trees [6-8].

Bodies of them are usually empty, often make a good progress. Presence of the secretoranbars in all parts is very specific. Johrenia paucijuga blossoms in June in the western region of Azerbaijan (Gedebey region). For now, no research is available on the biological advantages of this family and its biochemistry. Current research is devoted to study the chemical composition of Johrenia paucijuga.

The research materials and methods

As an object of study is taken the body of the Johrenia paucijuga in the flowering phase (end of June) in the Gadabey region of Azerbaijan. The substance was obtained by finely cutting of the Johrenia paucijuga body, then drying in the room conditions (300 g) and extracting by acetone 3 times (3 days for each time). Column chromatography method was used to get the individual items. Thus, the sum of 10 g ingredients was chromatographed in the glass column (h=60.0; d=2.5 cm) filled with Al₂O₃ (neutral, with III-IV degrees of activity). The volume of each fraction was taken 100 ml. The column chromatography was eluated by Hexane (36 fractions), hexane+benzolla (43 fractions), benzolla (41 fractions), benzolla+xloroforumla (28 fractions), xloroforumla (5 fractions) and xloroforum+alcohol (95: 5) [9].

The individuality of the substances was defined using thin layer chromatography method (Silufol UV 254, solvent-benzol+xloroforum 1:1), the melting temprature on the Boytius table.

IR-spectrums were drawn in the UR-20 spectrometer on the vaseline oil.

¹H, ¹³C, ¹³C Dept 135, Dept 90 spectrums were registered in the Bruker 300 MHz spectrometer for the isotope ¹H with frequency 300 MHz and for the ¹³C isotope with 75 MHz frequency using DMSO-d₆ as a solvent. Chemical replacements are given in the S-scale (internal standard-TMS).

Two individual substances were obtained from the plant material mix using chromatography method. The characterizations of the β-sitosterol and β-stigmasterol were defined on the basis of discovering the IR and NMR spectrums of the substances. The substance was obtained from the 73-75-th fractions by deluation by the (2:1) mixture of the chromatographic column. The NMR data of β-sitosterol (a) and β-stigmasterol (b) are presented in Tables 1 and 2.

(a)
1HNMR	13C NMR
1	37.23
2	31.6
33.52 (m, 1H)	71.83
4	42.26
5	140.72
65.34 (s, 1H)	121.74
7	31.8
8	31.8
9	50.11
10	36.15
11	21.07
12	39.26
13	42.26
14	56.75
15	24.30
16	28.25
17	56.03
180,68 (s, 3H)	11.86
191.01 (s, 3H)	19.40
200,92 (d, J=6.5 HZ, 3H)	35.88
21	18.25
22	33.80
23	26.02
24	45.81
25	29.11
26           0.83 (m, 3H)	19.83
27           0.84 (m, 3H)	19.02
280,84 (m, 2H)	23.04
290,84 (m, 3H)	11.98
1. beta-sitosterol

Table 1: ¹H NMR and ¹³C NMR chemical shift values for β-sitosterol (a) recorded in CDCl₃.

(b)
1HNMR	13C NMR
1	37.23
2	31.6
33.51 (tdd,1H, J=4.5,4.2 and 3.8Hz)	71.83
4	42.26
531 (t,1h,J=6.1 Hz)	140.72
6	121.74
7	31.8
8	31.8
9	50.11
10	36.15
11	21.07
12	39.66
13	42.26
14	56.85
15	24.36
16	28.24
17	55.93
180.71 (s, 3H)	12.04
191.03 (s, 3H)	19.40
20	40.52
210.91 (d, 3H, J=6.2 Hz)	21.21
224,98 (m, 1H)	138..33
235,14 (m, 1H)	129.25
24	51.24
25	31.8
260,82 (d, 3H,J=6.6 Hz)	19.07
27   0.80(d, 3H, J=6.6 Hz)	18.77
28	25.42
290.83 (t, 3H, J=7,1 Hz)	12.26
2.beta-stigmasterol

Table 2: ¹H NMR and ¹³C NMR chemical shift values for β-stigmasterol (b) recorded in CDCl₃.

The isolated compounds were identified as β-sitosterol and β-stigmasterol. The two formers are phytosterol with chemical structure similar to cholesterol. They reduce blood levels of cholesterol and are sometimes used in treating hypercholesterolemia. β-stigmasterol may be useful in prevention of certain cancers, including ovarian, prostate, breast and colon cancers [10-13].

Further research is recommended for isolation of other phytochemicals and also investigation of pharmacological and biological properties.

Two individual substances were obtained from the plant material mix using chromatography method. The characterizations of the β-sitosterol and β-stigmasterol were defined on the basis of discovering the IR and NMR spectrums of the substances. The isolated compaunds were identified as β-sitosterol and β-stigmasterol. The two formers are phytosterol with chemical structure similar to cholesterol. They reduce blood levels of cholesterol and are sometimes used in treating hypercholesterolemia. β-stigmasterolmay be useful in prevention of certain cancers, including ovarian, prostate, breast and colon cancers.