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Showing 5 results for Chromatography

Roya Karamian, Fatemeh Ghasemlou,
Volume 1, Issue 2 (3-2015)
Abstract

Saponins are secondary metabolites that are found in many plants and some animals. These compounds are high molecular weight glycosides, consisting of a sugar moiety linked to a triterpene or steroid aglycone. Many saponins have detergency properties and give stable foam in water. The genus Silene L. with more than 700 species is one of the largest genera of the family Caryophyllaceae mainly distributed in northern hemisphere. Saponins are one of the important secondary metabolites in the members of the genus. Quantitative and qualitative study of the saponins in the aerial parts and roots of three Silene species, namely S. ginodioica Ghaz. subsp. Penducularis (Fenzl ex Boiss.) Melzh., S. spergulifolia (Willd.) M. Bieb. and S. swertiifolia Boiss. were carried out by spectrophotometry and TLC methods. In this study, 5 fractions were obtained from each plant part and in both parts of the three species, fraction 1 contains the highest amounts of saponins. On the other hands, root extracts have high amounts of saponins. In addition, thin layer chromatography (TLC) of the extracts revealed some saponins spots with different Rf.


Seyyedeh Madjideh Mohammadnejad Ganji, Hossein Moradi, Ali Ghanbari, Mohammad Akbarzadeh,
Volume 4, Issue 2 (9-2017)
Abstract

Secondary metabolites have great pharmaceutical value in medicine. This study evaluates the effect of alti-tude on the amount and variety of secondary metabolites in lavender plant from two sites located in Mazandaran prov-ince. Sampling was performed in a randomized design with three replications. After the plants were dried, essential oil was extracted by distillation. Isolation and identification of constituents of the oil were performed by gas chromato-graphy and gas chromatography connected to mass spectroscopy. The results of this research led to the identification of 22 (99.96% of the total constituents) and 24 (97.01% of the total constituents) constituents in Baladeh and Behshahr sa-mples, respectively. Characteristics constituents included Andoyorneol (22.36%), 1, 8 cineole (20.7%), camphor (8.69%), α-Cadinol (7.60%), Caryophyllene oxide (5.09%) and Propanol (18.4%) in Behshahr population samples and Borneol (26.78%), 1, 8 cineole (20.19%), camphor (9.59%), α-Cadinol (5.80%), caryophyllene oxide (4.99%) and pr-opanol (3.41%) in Baladeh population samples. The differences in the output and combination of the essential oils in-dicated that the environmental as well as genetic factors can affect the production and amounts of the chemical comp-ounds in medical plants. Therefore, medicinal plants should be cultured in proper areas based on the aim of cultivation and the desired type of active materials.
Seyedeh Mahdieh Sadadt, Zahra Hajihassan, Mohammad Barshan-Tashnizi, Mehri Abdi,
Volume 5, Issue 3 (12-2018)
Abstract

Nerve growth factor (NGF) is a neurotrophic factor that is functional in the survival, maintenance and differentiation of nervous system cells. This protein has three subunits, of which the beta subunit has the main activity. Its structure consists of a cysteine knot motif made up of beta strands linked by disulfide bonds. It can be used as a therapeutic agent in the treatment of many diseases. As NGF extracted from natural sources is unsuitable for therapeutic goals, many studies have attempted to produce recombinant β-NGF. In this study, Trigger Factor (TF) chaperone was expressed simultaneously with β-NGF in E. coli in order to obtain increased yield of soluble recombinant human β-NGF.  For this purpose, pET39b (+)::β-NGF and chaperone plasmid pTf16 were transferred to E.coli (DE3 strain). After the induction of each promoter, the total proteins and periplasmic proteins were extracted. To confirm the effects of TF on total protein and soluble β-NGF expression level, Bradford and Dot blot techniques and ImageJ software were used. Then, β-NGF was purified using affinity chromatography column (Ni+2-NTA). Also, the PC12 cells were treated with the protein for one week in order to study the function of purified NGF. Our data indicated that co-expression of TF could increase the soluble and periplasmic production of β-NGF but not total proteins. Also, the treatment of PC12 cell line with purified β-NGF, co-expressed with TF chaperone, showed differentiation of these cells to nerve cells. This indicated that the purified NGF is fully functional. Our data suggest that the co-expression of cytoplasmic chaperone (TF) with recombinant nerve growth factor might be an efficient approach to produce a proper quantity of soluble and active rhNGF.


Razieh Sadat Solouki Nezhad, Hanieh Asaadi, Yaser Eshaghi Milasi, Sajjad Yazdansetad,
Volume 6, Issue 1 (5-2019)
Abstract

The production of pigments from bacteria is significant due to the low cost, high yield and ease of extract compared with other sources. Carotenoids are one of the most important pigments with antioxidant properties which are the precursor of vitamin A synthesis and have antibody overproduction ability, anti-tumor activity and inhibitory effect on the cardiovascular disease. The present study aimed to isolate and identify carotenoid-producing bacteria by high-performance liquid chromatography (HPLC) analysis of their carotenoid pigments. Twenty soil samples were collected from different regions of Tehran. After serial dilution each sample was cultured on BHI agar medium and incubated at 37°C. The pigment-producing bacteria were selected for further identification and their pigments were extracted by methanol. The screening was carried out at two levels: i) selection of the strains by visual color inspection, ii) analysis of the pigment extracts by UV-VIS spectroscopy and HPLC. The isolates were identified by phenotypic methods and their 16S rDNA gene was amplified by PCR method and sequenced. Staphylococcus epidermidis, Micrococcus aloeverae, Citricoccus alkalitolerans, Rhodococcus zopfii, Arthrobacter agilis, Dietzia natronolimnaea and Rhodococcus ruber were identified as carotenoid-producing strains. The highest rate of absorption was observed using UV-VIS analysis in Staphylococcus epidermidis and Dietzia natronolimnaea. The comparison of HPLC analysis with the standard β-carotene curve showed that the carotenoids were beta-carotene. Micro-organisms are a potential source in the production of pigments. In this study we introduced two genera of bacteria (Staphylococcus epidermidis and Dietzia natronolimnaea) with carotenoid-producing ability.
 
 


Saeideh Khamushi, Fatemeh Nejadhabibvash,
Volume 11, Issue 1 (6-2024)
Abstract

     The aim of this study was to evaluate the changes in the essential oil chemical composition of Satureja bachtiarica Bunge were inoculated by spores of the arbuscular mycorrhizal inoculation Glomus caledonium and foliar application of zinc chelate levels (0, 2, 4 and 8 g/L) in 2019, at Zarrin Giah greenhouse of Urmia city in West Azerbaijan Province, in a randomized complete block design with three replications. Plants were harvested at full flowering stage. Essential oils were obtained by Clevenger apparatus and were analyzed by GC and GC/MS. Comparison of means by Duncan's multiple range test showed that inoculation with Glomus caldonium had no significant effect on the essential oil yield of Satureja bachtiarica flowering shoots, but the foliar application of zinc chelate had statistically significant effect. However, there was no difference among different concentrations of zinc chelate in terms of effect on essential oil yield. The treatments also changed the essential oil composition compared to the essential oil of the control group. These changes included the reduction of beta-caryophyllene, sabinen hydrate, borneol and carvacryl acetate compounds and the increase of citral and octyl phthalate in the plant inoculated with mycorrhiza, as well as the increase of carvacrol in the essential oil of plants treated with zinc chelate.  Also, inoculation with mycorrhizal fungi and zinc chelate foliar application increased the amount of sesquiterpene & diterpene and monoterpene compounds, respectively. 

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