Microscopic characterization, TLC fingerprinting and optimization of total lipid content from Euphorbia neriifolia (L.) using response surface methodology.

Euphorbia neriifolia (EN) is a medicinal plant used to treat a variety of ailments in traditional systems. Despite numerous studies on pharmacological activities, no information was available on the microscopic study of this plant. This is the first study that has been attempted to fill this need by performing the light and field emission scanning electron microscopy (FESEM) of leaf, stem, and latex. The powder microscopy of several organs (leaves, stem, and bark) and exudate (latex) of EN was carried out using safranine, fast green, phloroglucinol, and other standard solutions at different magnifications. The chemical fingerprinting of petroleum ether extract was accomplished by using thin layer chromatography. The optimization of total lipid content from the EN leaf under ultrasound-assisted extraction (UAE) and soxhlet extraction (SE) procedure was determined using response surface methodology (RSM). The studied factors that affect the lipid content were: solvent ratio, extraction temperature, and extraction time. Several notable characteristics observed in the leaf of EN are amphistomatic leaves with anticlinical cell walls, anomocytic stomata, spongy mesophyll cells, elongated palisade cells, angular collenchyma, and U-shaped vascular bundle. The plano-convex midrib is covered by polygonal to oval-shaped cuticles and contains anomocytic stomata. The circular petiole has no trichomes and contains laticifers, crystals, and idioblasts. The circular stem was observed with trichomes, hypodermis, collenchyma, parenchymatous cells, central pith, pentagonal stellar region, cambium, and 2-4 times more xylem that of phloem. All of the powdered plant parts and exudate under study contained trichomes, xylem vessels, wood fibers, cork cells, starch grains, calcium oxalate crystals, idioblasts, lignified cork, tannin content, stone cells, and oil globules. The blackish-green colored petroleum ether extract with semi-solid consistency showed the greatest percent (%) yield of 4% in the latex of EN. The thin layer chromatography (TLC) examination of petroleum ether extract of EN leaf produced a maximum 6 spots with Rf values of 0.16, 0.58, 0.62, 0.73, and 0.96 in the mobile phase of petroleum ether-acetone (8:2). In terms of optimization, the dark green colored UAE extract with semi-sticky consistency showed highest % yield of 4.5% whereas the yellowish green colored SE extract of sticky consistency showed the highest % yield of 4.9%. The findings showed that there were not many differences in the total lipid content between UAE (0.16%) and SE (0.11%). However, the best optimum condition for lipid content extraction analysis was obtained as follows: solvent ratio (PE:HE) 50:50, extraction temperature 50°C, extraction time 45 min for UAE, and solvent ratio (PE:HE) 60:40, extraction temperature 45°C, and extraction time of 24 h for SE. Hence, this study signifies the various noteworthy microscopic features along with the presence of different phytocompounds through TLC and best optimized condition for the extraction of lipids from different parts of EN. As no previous study has been reported, the outcomes obtained from the current study prove to be beneficial in the identification of species, quality control, and detection of any adulteration from the laboratory and commercial samples of EN. RESEARCH HIGHLIGHTS: The percent yield was found to be maximum in latex extract (4%). The leaf pet ether extract was separated into 6 bands with different Rf values. The extracted compounds from Euphorbia neriifolia leaves were categorized into non-polar heat tolerant. The highest total lipid yield (0.1119) was obtained at solvent ratios 60:40 of PE:HE (petroleum ether: petroleum hexane).

Structural characterisation of bioactive compounds of Gymnosporia senegalensis (Lam.) Loes. using advanced analytical technique like FT-IR, GC-MS and 1H-NMR spectroscopy.

The present investigation was carried out to characterise bioactive components from G. senegalensis by using Fourier-transform infra-red (FT-IR) spectroscopy, 1H-nuclear magnetic resonance spectroscopy, and gas chromatography-mass spectrometry (GC-MS). The FTIR analysis confirmed the presence of > CH2, -CH3, C = C-C, C-H, C-F, C = C, -C = N-, C-C = N-, and -OH functional groups. The 1H-NMR spectrum revealed the presence of structures of four bioactive compounds i.e. tetratetracontana derivative, ß-carotene, amyrin, and terpineol. GC-MS revealed the presence of different types of high and low molecular weight chemical entities with varying quantities including volatile and essential oil, monoterpenoid, tetraterpenoid, carotenoid, terpenoid, triterpenes, and nortriterpenes. From the results, it could be concluded that G. senegalensis contains various bioactive compounds of biological and pharmacological importance. Overall, this study will provide insight into the characterisation and development of drugs from medicinal plants.

Isolation, development and validation of HPTLC method for the estimation of ß-carotene from Gymnosporia senegalensis (Lam.) Loes.

The present study is aimed to isolate terpenoids from Gymnosporia senegalensis through analytical and preparative thin-layer chromatography (TLC) and to determine their antioxidant activity using the 2, 2-diphenyl-1- picrylhydrazyl (DPPH) assay and to find out the presence of ß-carotene through high-performance thin-layer chromatography (HPTLC). The validation included linearity, limit of detection (LOD), limit of quantification (LOQ), specificity, precision, recovery, and robustness. All the isolated compounds from TLC exhibited significant antioxidant activity. Among all, isolated compounds from leaf showed highest IC50 values. The highest total terpenoid content (TTC) was found 51.6 ± 0.06 in stem, then 49.02 ± 0.01 in bark, and 46.27 ± 0.01 in leaf. DPPH results indicated that leaf-isolated compound 1 (LIC1) showed the highest IC50 at 7.55 ± 0.02 and stem-isolated compound 2 (SIC2) showed the lowest IC50 at 0.616 ± 0.01 among all the isolated compounds of G. senegalensis. HPTLC separation was carried out on aluminium plates pre-coated with silica gel 60 F254 as the stationary phase and n-hexane: ethyl acetate (6:4, v/v) as the mobile phase. Quantification was achieved based on a densitometric analysis of ß-carotene in the concentration range of 100-500 ng/band at 254 nm. For the calibration plots, linear regression produced r2 = 0.96450 and Rf = 0.27. The LOD and LOQ were 10.15 and 30.76 ng/mL for HPTLC and relative standard deviation were 137.26 ± 2.03 and 160.43 ± 2.95 (intra-day) and 127.88 ± 2.14 and 157.27 ± 1.90 (inter-day) for 200 and 400 ng/band, respectively. The present study shows the presence of various types of terpenoids through TLC whereas the HPTLC results indicated that the developed methods were accurate and precise. It also shows that the approach is appropriate for its intended use in routine quality control testing of commercially available tablet formulations and drug assay to assist both industries and researchers in making important decisions at a reasonable cost. Moreover, due to the use of a safer and more environmentally friendly mobile phase in comparison to the toxic mobile phases used in recent analytical techniques to estimate ß-carotene, this methodology is also secure and sustainable.

Bioprospecting of endophytes in medicinal plants of Thar Desert: An attractive resource for biopharmaceuticals.

Endophytes live asymptomatically within the healthy tissues of plant parts of the host, has grabbed the attention of ecologists, chemists, and researchers to have a broad spectral of biotechnological potential. It has been proven that almost all plants harbor endophytes within themselves. Numerous studies indicated that endophytes act as chemical synthesizers of the secondary metabolites of their host plant. Various medicinal plants of the Thar Desert have been used by the local folks of the Rajasthan to treat several diseases ailments for time immemorial. On the basis of their prior knowledge of ethnopharmacological usage of medicinally important plants of Thar Desert, several researchers directed their studies in search of endophytic microflora of such medicinally important plants for the discovery of novel bioactive molecules of pharmaceutical importance, for instance, taxol producing endophytic fungus Phoma sp. isolated from Calotropis gigantea as well as Aspergillus fumigatus, an endophytic fungus reported from Moringa oleifera demonstrated an effective antibiofilm, antimicrobial and antiproliferative activity. This review sheds light on the endophytic microflora of the ethnomedicinal plants of the Thar Desert and their biopotential as a promising source of pharmaceutically important naturally derived compounds.

Metabolic Profile, Bioactivities, and Variations in the Chemical Constituents of Essential Oils of the Ferula Genus (Apiaceae).

The genus Ferula is the third largest and a well-known genus of the Apiaceae family. It is categorized in the Peucedaneae tribe and Ferulinae subtribe of the Apiaceae family. At present, about 180 Ferula species have been reported. The genus is mainly distributed throughout central and South-West Asia (especially Iran and Afghanistan), the far-East, North India, and the Mediterranean. The genus Ferula is characterized by the presence of oleo-gum-resins (asafoetida, sagapenum, galbanum, and ammoniacum) and their use in natural and conventional pharmaceuticals. The main phytochemicals present in the genus Ferula are as follows: coumarin, coumarin esters, sesquiterpenes, sesquiterpene lactones, monoterpene, monoterpene coumarins, prenylated coumarins, sulfur-containing compounds, phytoestrogen, flavonoids and carbohydrates. This genus is considered to be a valuable group of medicinal plants due to its many different biological and pharmacological uses as volatile oils (essential oils). Numerous biological activities are shown by the chemical components of the essential oils obtained from different Ferula species. Because this genus includes many bioactivities such as antimicrobial, insecticidal, antioxidant, cytotoxic, etc., researchers are now focusing on this genus. Several reviews are already available on this particular genus, including information about the importance and the uses of all the phytochemicals found in the species of Ferula. Despite this, no review that specifically provides information about the biological activities of Ferula-derived essential oils, has been published yet. Therefore, the present review has been conducted to provide important information about the chemical profile, factors affecting the chemical composition, and biological activities of essential oils of the Ferula species.