Fatty Acid Composition and Phytosterols from Pyrola Rotundifolia
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FATTY ACID COMPOSITION AND PHYTOSTEROLS FROM Pyrola rotundifolia
I. G. Nikolaeva,1,2* L. P. Tsybiktarova,2 Zh. A. Tykheev,2,3 V. V. Taraskin,2,3 L. D. Radnaeva,2,3 and G. G. Nikolaeva1,2
Pyrola rotundifolia L. (Pyrolaceae) is a perennial herbaceous plant distributed over the Arctic, Europe, Caucasus, western and eastern Siberia, the Far East, China, and Mongolia [1, 2]. Aqueous and EtOH extracts and tincture of leaves exhibited antibacterial, immunomodulating, antioxidant, tonic, and diuretic activity [1–3]. Infusions, decoctions, and tincture of the aerial part of the plant are used in Tibetan medicine for liver diseases and bone tuberculosis and as a cholagogue and sedative [2]. Leaves of the plant are used in folk medicine for rheumatism, stomachache, hemorrhage, purulent kidney diseases, and female diseases, especially those associated with inflammation [1, 4]. The lipid composition of this plant has not been previously studied. The aerial part of P. rotundifolia collected during flowering (July 2019) in Pribaikalsky District, Republic of Buryatia, was studied. The quantitative content of the lipid fraction was preliminarily determined [5]. The method was based on extraction of lipids by petroleum ether in a Soxhlet apparatus. The content of the lipid fraction from the aerial part was 1.43% of the air-dried raw material mass (average of three determinations). Total lipids were isolated by a modified Bligh–Dyer method [6]. Raw material was extracted by CHCl3–MeOH (1:2, v/v). The mixture was homogenized (2×) and filtered. The filtrate was treated with CHCl3 and H2O. The CHCl3 layer was diluted with C6H6 and concentrated under vacuum. Fatty acid methyl esters (FAME) were prepared using HCl (2 N) in MeOH for 2 h at 90°C. FAME were extracted (3×) with hexane. The extract was evaporated and treated with bis(trimethylsilyl)trifluoroacetamide-N,O (BSTFA) to produce volatile derivatives of sterols and free acids (45 min at 80°C). The obtained reaction mixture was dissolved in hexane and analyzed by GC-MS on an Agilent Packard HP 6890 gas chromatograph with an HP MSD 5973N quadrupole mass spectrometer as the detector. FAME were chromatographed using a column (30 m × 0.25 mm × 0.25 μm) of HP-5ms (copolymer of 5% diphenyl- and 95% dimethylsiloxane), He carrier gas (constant flow rate 1.5 mL/min), column temperature 125°C (isothermal for 0.5 min), 125–320°C (7°C/min, isothermal for 0.5 min), vaporizer temperature 280°C, ion source 230°C. The sample volume was 1 μL with 40:1 flow division. The percent composition of the mixture was calculated from gas chromatographic peak areas without using correction factors. Qualitative analysis was based on comparison of retention times and total mass spectra of the corresponding pure compounds using the NIST11.L library and standard mixtures (Bacterial Acid Methyl Esters, CP Mix, Supelco, Bellefonte, PA, USA; FAME mixture, Supelco 37 comp. FAME Mix, 10 mg/mL in CH2Cl2). A total of 22 compounds were identified in the sample. Table 1 presents the GC-MS analytical results and shows const
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