The GC-MS analysis of methanolic extract of Chlorophytum borivilianum and compounds’ activities validation at standard databases

Chlorophytum borivilianum (Safed Musli), a member of the Liliaceae family is a well-known plant in India for its aphrodisiac and adaptogenic activities. The present study was carried out to identify the novel compounds based new medicinal properties of the plant. The preliminary phytochemical screening and GC-MS based chromatographic analysis of a methanolic extract of C. borivilianum (MECB) tubors resulted the detection of Phenol, Tannins, Flavo-noids, Alkaloids, Saponins, and Glycosides in the plant. In GC-MS analysis of the MECB, more than 200 compounds were identified in different peak are-as. All of them were crosschecked at various online compound databses like PubChem, Drugbank, etc. for the identification of their known biological ac-tivities, if any. Out of them, mainly 21 compounds (falling in 70.74% peak area) like 4-mercaptophenol, 1,4-Benzenediol, 2-methoxy-, Octadecanal, Pentadecanal, 9,12-Octadecadienoic acid, Betulin, Levomenthol, etc. were characterized in this study. As per their reported medicinal activities at standard databases the plant tubor is hereby reported to have anti-microbial, analgesic, anti-inflammatory, anti-neoplastic, anti-pruritic, anti-tussive and anti-spasmodic activites. Moreover, the compounds like Cyclo-hexanol, 5-methyl-2-(1-methylethyl), Palmitoyl Chloride, Triarachine, Phytol, 4-mercaptophenol with their mentholating, food additives and other valua-ble effects have underlined the industrial value of the plant.

Intestinal microbe-dependent ω3 lipid metabolite αKetoA prevents inflammatory diseases in mice and cynomolgus macaques

Dietary ω3 fatty acids have important health benefits and exert their potent bioactivity through conversion to lipid mediators. Here, we demonstrate that microbiota play an essential role in the body’s use of dietary lipids for the control of inflammatory diseases. We found that amounts of 10-hydroxy-cis-12-cis-15-octadecadienoic acid (αHYA) and 10-oxo-cis-12-cis-15-octadecadienoic acid (αKetoA) increased in the feces and serum of specific-pathogen-free, but not germ-free, mice when they were maintained on a linseed oil diet, which is high in α-linolenic acid. Intake of αKetoA, but not αHYA, exerted anti-inflammatory properties through a peroxisome proliferator-activated receptor (PPAR)γ-dependent pathway and ameliorated hapten-induced contact hypersensitivity by inhibiting the development of inducible skin-associated lymphoid tissue through suppression of chemokine secretion from macrophages and inhibition of NF-κB activation in mice and cynomolgus macaques. Administering αKetoA also improved diabetic glucose intolerance by inhibiting adipose tissue inflammation and fibrosis through decreased macrophage infiltration in adipose tissues and altering macrophage M1/M2 polarization in mice fed a high-fat diet. These results collectively indicate that αKetoA is a novel postbiotic derived from α-linolenic acid, which controls macrophage-associated inflammatory diseases and may have potential for developing therapeutic drugs as well as probiotic food products.

GC-MS ANALYSIS PROFILE AND BIOACTIVE COMPONENTS OF FLOWERS OF Bergenia ciliata, A WEED OF ROCK CREVICES

Bergenia ciliata is a weed of hilly areas of Pakistan that generally grows in rock crevices. In the present study, flowers of this weeds were collected from Murree. The dried flowers were extracted in methanol and subjected to GC-MS analysis that showed 7 compounds in it. The predominant compound was hexanedioic acid, bis(2-ethylhexyl) ester (48.88%) followed by γ-sitosterol (22.56%). Moderately occurring compound was cyclohexane, 1,3,5-triphenyl- (12.87%). The remaining compounds namely n-hexadecanoic acid (4.97%), pentadecanoic acid, 14-methyl-, methyl ester (3.77%), 9,12-octadecadienoic acid, methyl ester, (Z,Z)- (2.94%) and 9,12,15-octadecatrienoic acid, methyl ester, (Z,Z,Z)- (2.92%) were categorized as less abundant compounds. Literature survey indicated that these compounds possess antimicrobial, larvicidal, anti-inflammatory, anticancer and/or antidiabetic properties.

Developing a platform for production of the oxylipin KODA 9-hydroxy-10-oxo-octadecadienoic acid in plants

KODA (9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid) is a plant oxylipin involved in recovery from stress. As an agrichemical, KODA helps maintain crop production under various environmental stresses. In plants, KODA is synthesized from α-linolenic acids via 9-lipoxygenase (9-LOX) and allene oxide synthase (AOS), although the amount is usually low except in the free-floating aquatic plant Lemna paucicostata. To improve KODA biosynthetic yield in other plants such as Nicotiana benthamiana and Arabidopsis thaliana, we developed a system to overproduce KODA in vivo via ectopic expression of L. paucicostata 9-LOX and AOS. The transient expression in N. benthamiana showed that the expression of these two genes is sufficient to produce KODA in leaves. However, stable expression of 9-LOX and AOS (with consequent KODA production) in Arabidopsis plants succeeded only when the two proteins were localized in plastids or the endoplasmic reticulum/lipid droplets. Although only small amounts of KODA could be detected in leaf extracts of transgenic Nicotiana or Arabidopsis plants, subsequent incubation of the extracts increased KODA abundance over time. Therefore, KODA production in transgenic plants stably expressing 9-LOX and AOS requires specific subcellular localization of these two enzymes and incubation of leaf crude extracts, which liberates α-linolenic acid via breakdown of endogenous lipids.

Soxhlet Extraction versus Hydrodistillation Using the Clevenger Apparatus: A Comparative Study on the Extraction of a Volatile Compound from Tamarindus indica Seeds

The present study aims to compare two traditional extraction techniques. A volatile compound from Tamarindus indica seed was obtained with Soxhlet extraction (SE) and hydrodistillation using the Clevenger apparatus (HDC). The extraction yield and chemical composition of the essential oil samples were compared. Both oils extracted were analyzed with GC-MS, and forty-one chemical compounds were identified in essential oil components from SE while forty chemical compounds were found in the HDC-extracted oil sample. The major essential oil components present in both the SE and HDC method are cis-vaccenic acid, 2-methyltetracosane, beta-sitosterol, 9,12-octadecadienoic acid (Z, Z)-, and n-hexadecanoic acid in varying concentrations. Moreover, the essential oils obtained by both methods look similar quantitatively but differ qualitatively. The HDC method produced more oxygenated compounds that contribute to the fragrance of the oil. The major constituents observed in the essential oil extracted by SE methods include cis-vaccenic acid (17.6%), beta-sitosterol (12.71%), 9,12-octadecadienoic acid (Z, Z)- (11.82%), n-hexadecanoic acid (8.16%), 9,12-octadecadienoic acid, methyl ester (5.84%), oleic acid (4.54%), and 11-octadecenoic acid and methyl ester (3.94%). However, in the hydrodistillation technique, the oil was mostly composed of 9,12-octadecadienoic acid (Z, Z)- (23.72%), cis-vaccenic acid (17.16%), n-hexadecanoic acid (11.53%), beta-sitosterol (4.53%), and octadecanoic acid (3.8%). From the data obtained, HDC seems to be a better method for extraction of Tamarindus indica essential oil compared to the Soxhlet extraction apparatus.

The Linoleic Acid Content of the Stratum Corneum of Ichthyotic Golden Retriever Dogs Is Reduced as Compared to Healthy Dogs and a Significant Part Is Oxidized in Both Free and Esterified Forms

Golden Retrievers may suffer from Pnpl1-related inherited ichthyosis. Our study shows that in the stratum corneum (SC) of ichthyotic dogs, linoleic acid (LA) is also present in the form of 9-keto-octadecadienoic acid (9-KODE) instead of the acylacid form as in normal dogs. The fatty acids purified from SC strips (LA, acylacids) were characterized by liquid chromatography-tandem mass spectrometry (LC-MS) and atmospheric pressure chemical ionization (APCI). Electrospray ionization (ESI) and MS2(MS/MS Tandem mass spectrum/spectra)/M3 (MS/MS/MS Tandem mass spectrum/spectra) fragmentation indicated the positions of the double bonds in 9-KODE. We showed that ichthyotic dogs have a threefold lower LA content in the form of acylacids. The MS2 fragmentation of acyl acids showed in some peaks the presenceof an ion at the m/z 279, instead of an ion at m/z 293 which is characteristic of LA. The detected variant was identified upon MS3 fragmentation as 9-keto-octadecadienoic acid (9-KODE), and the level of this keto-derivative was increased in ichthyotic dogs. We showed by the APCI that such keto forms of LA are produced from hydroperoxy-octadecadienoic acids (HpODE) upon dehydration. In conclusion, the free form of 9-KODE was detected in ichthyotic SC up to fivefold as compared to unaffected dogs, and analyses by HPLC (High performance liquid chromatography) and ESI-MS (Electrospray Ionization-Mass Spectrometry) indicated its production via dehydration of native 9-HpODE.

GC-MS, FTIR and Antiulcer Screening of Aqueous Seed Extract and Oil of Nigella sativa in Wistar Rats

Peptic ulcer disease (PUD) is a sore in the lining of the stomach or duodenal mucosa. This study was aimed at evaluating the antiulcer activity of the aqueous extract of Nigella sativa (black seed) and its constitutents. The pharmacognostic properties of the dry seeds were determined. The seeds were extracted using two methods digestion and hydro-distillation. The acute toxicity, phytochemical constituents and the antiulcer evaluation were performed on ethanol-induced ulcer in wistar rats using a standard method. FTIR and GC-MS analysis of the aqueous seed extract was also determined using standard methods. Nigella sativa seed shows the presence of starch grains, lignified tissues, tannins, cellulose, protein and oil globules. The aqueous extract has a high safety margin. The phytochemical studies revealed the presence of saponins, flavonoids, alkaloids, tannins, glycosides, fats and oil. The black seed aqueous and oil extract at 500 mg/kg significantly reduced the acidity, total acidity, and ulcer index, and pH of gastric content when compared with the positive control (Famotidine). The FTIR analysis identified the presence of the following functional groups chloro, ether, amine, carboxylic acid, nitriles, methylene, alcohol, while the GC-MS identified five compounds such as glycerin, n-Hexadecanoic acid, 9, 12-octadecadienoic acid-methyl ester, 9, 12-octadecadienoic acid and 9, 12-octadecadienoyl chloride. The pharmacognostic properties can act as a reliable tool for the standardization of the plant part. This study suggests that aqueous and oil extract possess antiulcer properties. Thus the aqueous and oil extract of black seed can be considered as antiulcer medication traditionally. Keywords: Nigella sativa, Black seed, Pharmacognostic, Phytochemical, Acute toxicity (LD50), Antiulcer.

Sorghum extract: Phytochemical, proximate, and GC-MS analyses

Introduction. Sorghum is available cereal seeds of African origin belonging to the Poaceae family. However, its metabolites and proximate composition have not studied well, which led to the under-utilization of this cereal. This research aimed to investigate the classes of phytochemical and proximate compositions of sorghum extract in order to assess its nutraceutical potential for food chemistry and dietary formulations. Study objects and methods. We studied the sorghum seed oil extract obtained with the help of a Soxhlet extractor. Sorghum was purchased in Ota, Nigeria. The bioactive compounds were identified by standard methods of phytochemical screening, the nutritional content was investigated with proximate analysis, and the secondary metabolites in the sorghum extract were determined using gas chromatography – mass spectrometry (GC-MS). Result and discussion. The phytochemical screening showed the presence of steroids, saponins, terpenoids, alkaloids, cardiac glycosides, and quinones in the sorghum extract. The oil yield obtained was 11.00 ± 0.18%. The proximate analysis revealed 5.94% moisture content, 3.05% ash, 0.20% crude fiber, 11.00% fat, 5.54% protein, and 74.27% carbohydrates. The selected physicochemical parameters measured in the sorghum extract included cloud point (0.40°C), specific gravity at 25°C (0.81), and refractive index (1.46). The GC-MS analysis revealed the presence of 9,12-octadecadienoic acid (Z,Z)-, stigmasterol, 8-dodecen-1-ol, acetate, (Z)-, vitamin E, linoleic acid ethyl ester, and 9,12-octadecadienoic acid, methyl ester, which accounted for about 85% in the sorghum composition. Other constituents, presented at lower amounts, included 12-heptadecyn-1-ol, 1H-Imidazole-5-ethanamine, 1-methyl-, and cyclononene. Conclusion. The findings of this study revealed high nutritive potential of sorghum, which make it a rich source of energy for humans and animals.

Chemical Datasets, Antioxidant, Free Radicals Scavenger activities estimate in Aqueous Garlic (Allium sativum) extract

Garlic is one of many medicinal plants that used over thousands of years due to its organosulfur compounds. Materials and methods: commercial garlic was used for the extraction, phytochemicals detections and Gas Chromatography-Mass (GC- Mass) analysis utilized to determine the main compounds in the aqueous extract. Antioxidant activity was evaluated in the garlic extract using total antioxidant capacity (TAC) and DPPH for detecting the concentration that takes half of the free radicals. The phytochemical tests revealed several compounds in the extract, included alkaloid, glycosides, tannins, and flavonoid and phenol while saponin compound was not detected. GC-Mass spotted 15 different compounds included allyl trisulfide with the concentration (1.89%) followed by ascorbic acid 2, 6-dihexadecanoate (22.24%), 9, 12-Octadecadienoic acid (Z, Z) (11.44), and 6-Octadecenoic acid, (Z) (40, 28). Total Antioxidant Activity of garlic aqueous extract reached 60 mg/ml compared to that of ascorbic acid. IC50 examination observed that the aqueous extract of garlic had 308.57µg/ml value.

BIOCHEMICAL PROFILE OF Calotropis procera FLOWERS

Calotropis procera is a medicinal weed of family Asclepiadaceae. This study was carried out to explore the biochemical profile of C. procera flowers collected from Southern Punjab region of Pakistan. Methanolic flower extract of C. procera was subjected to GC-MS analysis. There were 30 compounds identified in this extract. The predominant compound was γ-sitosterol with 15.39% peak area. Other abundantly occurring compounds included stigmasterol (9.22%), 9,12-octadecadienoic acid (Z,Z)-, methyl ester (9.01%), campesterol (8.63%), α-amyrin acetate (8.25%), β-amyrin (8.09%), hexadecanoic acid, methyl ester (7.91%), 11-octadecenoic acid, methyl ester (6.15%), and 2-methoxy-4-vinylphenol (5.66%). Moderately abundant compounds included nonacos-1-ene (2.83%), methyl stearate (1.57%), pentacosane (1.44%), phytol (1.33%), heptacos-1-ene (1.20%), heneicosane (1.19%), and 1-hexacosene (1.09%). The remaining less abundant compounds were present with peak areas less than 1%. Literature survey showed that the major compounds identified in the flower extract of C. procera possess various bioactivities including antidiabetic, anticancer, antihyperglycemic, antioxidant, antimicrobial and anti-inflamatory.