Gas chromatography/mass spectrometry with chemometric methods for the determination of fatty acid profiles in herbal oils.

The conventional method for analyzing fatty acid is gas chromatography (GC) with polar capillary columns. However, owing to the high cost and the sensitivity of these columns to the presence of water and oxygen, these columns are not the best choice for separation. Also, analyzing long-chain acids (>C28) with polar columns is impossible. On the other hand, complete separation with nonpolar columns is not possible for some saturated, unsaturated and long-chain fatty acids. Therefore, in this study, with the help of chemometric methods, a method was developed using GC/mass spectrometry (MS) with a nonpolar column to resolve the peaks to completely separate and accurately identify and quantify fatty acids. Using this method, the fatty acid profiles of the seed oils of Sesamum indicum L, Nigella sativa, Pimpinella anisum, Linum asitatissimum L, Silybum marianum and Amygdalus communis L. var. Amara and var. Dulcis were identified. Through applying the multivariate curve resolution method after GC/MS, the C-18 fatty acids such as α-linolenic, linoleic, oleic and stearic acids were separated and quantified. Also, the total percentages of identified fatty acids increased by 1-6% after resolving overlapping peaks. Finally, the obtained percentages of saturated and unsaturated fatty acids were confirmed by reference reports.

Enhancement of Essential Oil Production and Expression of Some Menthol Biosynthesis-Related Genes in Mentha piperita Using Cyanobacteria.

Background: Mentha piperita L. is one of the most important aromatic crops and is cultivated worldwide for essential oils (EOs). Objectives: The aim of the present study was to investigate the potential of two cyanobacteria, Anabaena vaginicola ISB42 and Nostoc spongiaeforme var. tenue ISB65, as biological-elicitors to improve the growth and essential oil production of M. piperita. Materials and Methods: In this experiment, inoculation of M. piperita with cyanobacteria was performed by adding 1% cyanobacterial suspension to the soil of treated pots on the first time of planting and every 20 days thereafter. The experiment was performed in a randomized complete block design in an experimental greenhouse condition. After 90 days planting, the vegetative growth factors, the content of photosynthetic pigments, as well as the quantity and quality of EOs of treated and control plants were evaluated. Also, quantitative changes in the expression of some menthol biosynthesis-related genes were investigated. Results: Cyanobacterial application led to significant increases in M. piperita growth indices including root and shoot biomass, leaf number, leaf area, node number and ramification, as well as photosynthetic pigments content. The statistical analysis showed a 41-75 % increase in some of these growth indices, especially in Nostoc-treated plants. A. vaginicola and N. spongiaeforme var. tenue inoculation led to a 13% and 25% increase in the EOs content of M. piperita, respectively. The EOs components were also affected by cyanobacterial treatments. According to the statistical analysis, Nostoc-treated plants showed the highest amount of (-)-menthone and (-)-limonene, with a 2.36 and 1.87-fold increase compared to the control. A. vaginicola and N. spongiaeforme var. tenue inoculation also led to 40% and 98% increase in transcript level of (-)-limonene synthase gene, respectively. The expression of the (-)-menthone reductase gene, was also increased by 65% and 55% in response to A. vaginicola and N. spongiaeforme var. tenue application, respectively. Conclusions: Our data demonstrated that in addition to growth enhancement, these two heterocystous cyanobacteria improved the quantity and quality of EOs by up-regulating the key genes involved in the menthol biosynthetic pathway. Based on our results, these cyanobacteria can be considered valuable candidates in the formulation of low-cost and environmentally friendly biofertilizers in sustainable peppermint production.

Introduction of Adonis aestivalis as a new source of effective cytotoxic cardiac glycoside.

Cardiac glycosides are used for treatment of irregular heartbeats, cardiac arrhythmia and congestive heart failures. In this research, digitoxin as a cardiac glycoside was identified and isolated for the first time in the world from Adonis aestivalis and investigated for its cytotoxic activity against cervical cancer cell (HeLa) lines and human lymphocytes by MTT test. Digitoxin extracted from the aerial parts of the plant collected from west of Iran and purified by column and thin layer chromatographic techniques. The structure of isolated cardiac glycoside was identified by IR, 1H NMR and 13C NMR methods and so the presence of digitoxin was established. The half maximal inhibitory concentration values for cervical cancer and lymphocyte cells were obtained to be 5.62 and 412.94 µg/mL. The results of this study introduced the new resource of digitoxin which has considerable cytotoxic effects against HeLa cancer cells but did not damage normal human lymphocyte cells.

Antioxidant potential assessment of phenolic and flavonoid rich fractions of Clematis orientalis and Clematis ispahanica (Ranunculaceae).

The antioxidant activities of crude extract fractions using Hexane, Chloroform, Ethyl Acetate, Butanol and Water of Clematis orientalis and Clematis ispahanica were investigated. 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay and the ferric reducing/antioxidant potential (FRAP) were used to evaluate the antioxidant capacity. The total phenolics were found to be 4.37-9.38 and 1.32-11.37 mg gallic acid equivalents (GAE)/g in different fractions for C. orientalis and C. ispahanica, respectively. The ethyl acetate fraction of C. orientalis and chloroform fraction of C. ispahanica showed the highest DPPH and FRAP activities at a concentration of 300 µg/mL. The predominant phenolic compounds identified by HPLC in C. orientalis were Resorcinol (603.5 µg/g DW) in chloroform fraction and Ellagic acid (811.7 µg/g DW) in chloroform fraction of C. ispahanica.