Investigation of the presence of pharmaceuticals and personal care products (PPCPs) in groundwater of Jazan area, Saudi Arabia

Purpose: To investigate the possible occurrence of some selected  pharmaceutical compounds in the groundwater of Jazan area, Saudi Arabia.Methods: Water samples from 46 wells were collected from different sites covering Jazan area of Saudi Arabia between February and March 2017. These samples were first analyzed to investigate the presence of eleven drugs mostly used in the study area. Thereafter, samples were subjected to liquid chromatography-mass spectrometry (LC-MS/MS) by direct injection and external standard calibration.Results: Despite the low detection limit (0.001 - 0.02 μg/L) applied to the investigated compounds with a variety chemical groups (acetylsalicylic acid, paracetamol, ibuprofen, metronidazole, caffeine, olmesartan, omeprazole, nifedipine, diclofenac sodium, glibenclamide and loratidine), none of these compounds was detected in any of the analyzed samples.Conclusion: The main source of environmental contamination with  pharmaceuticals and personal care products (PPCPs) is wastewater. The results obtained reveal the absence of groundwater contamination by these compounds in Jazan area. However, further extended investigations and monitoring are recommended.Keywords: Pharmaceuticals, Groundwater, Wastewater, Pollution, Personal care products, Liquid chromatography-mass spectrometer (LC-MS/MS

Ultrafast monolithic HPLC method for simultaneous quantification of the anticancer agents, imatinib and sorafenib: Application to tablet dosage forms

Purpose: To develop and validate a simple ultrafast monolithic high performance liquid chromatography (HPLC) method for the simultaneous quantification of two anti-cancer agents, imatinib and sorafenib, in pure form and tablet preparations.Method: Chromatographic separation was accomplished using Chromolith flash RP-18 HPLC-column (25 - 4.6 mm; macropores, 2 μm; mesopores, 13 – 15 nm). The optimum mobile phase composition of ammonium acetate buffer (10 mM, pH 8.5) and methanol at ratio of 35:65 v/v was used. Effluent flow rate was adjusted to 1.0 mL/min and the analysis was performed at 250 nm wavelength. The developed method was evaluated for specificity, linearity, precision and accuracy.Results: The method offered a linear relationship over the concentration range of 1 - 16 μg/ml (correction coefficient, R2 = 0.9999) for both analytes. Limit of detection (LOD) was 0.1891 and 0.1888 μg/ml while limit of quantification (LOQ) was 0.6303 and 0.6294 μg/ml for imatinib and sorafenib, respectively. Mean recovery was within 100 ± 2 %. The utility of the new method was demonstrated by its successful use for the analysis of commercially available tablet formulations of both drugs.Conclusion: The developed method is fast and economical, and is being recommended for routine analysis of imatinib and sorafenib in bulk drug and tablet dosage forms in quality control laboratories.Keywords: RP-HPLC, Chromolith, Imatinib, Sorafenib, Validation, Quality contro

Comparing monolithic and fused core HPLC columns for fast chromatographic analysis of fat soluble vitamins

HPLC stationary phases of monolithic and fused core type can be used to achieve fast chromatographic separation as an alternative to UPLC. In this study, monolithic and fused core stationary phases are compared for fast separation of four fat-soluble vitamins. Three new methods on the first and second generation monolithic silica RP-18e columns and a fused core pentafluoro-phenyl propyl column were developed. Application of three fused core columns offered comparable separations of retinyl palmitate, DL-α-tocopheryl acetate, cholecalciferol and menadione in terms of elution speed and separation efficiency. Separation was achieved in approx. 5 min with good resolution (Rs >5) and precision (RSD ≤ 0.6 %). Monolithic columns showed, however, a higher number of theoretical plates, better precision and lower column backpressure than the fused core column. The three developed methods were successfully applied to separate and quantitate fat-soluble vitamins in commercial products

Antidepressant effect of methanol extract of smokeless tobacco and identification of its bioactive components

Purpose: To investigate the antidepressant effect of methanol extract of smokeless tobacco and identify its bioactive compounds. Methods: Adult Wistar rats were randomly assigned to five groups of five rats each: normal control group, standard (reference) control group as well as 100, 200 and 500 mg/kg extract group. The extract, standard drug (imipramine) and normal saline were administered via the intraperitoneal (i.p.) route. The rats were subjected to forced swim test (FST) and tail suspension test (TST) to assess the antidepressant effect of methanol extract of smokeless tobacco. Gas chromatography–mass spectrometry (GC-MS) was used to identify the bioactive compounds of the extract. Results: The oral LD50 of the extract was > 2000 mg/kg. Significant decrease in immobility time was observed after single administration of imipramine (p < 0.05). The extract significantly and dosedependently decreased the immobility time, but increased climbing and swimming times, when compared with normal control group (p < 0.05). The immobility time of stressed rats regardless of sex was significantly and dose-dependently lowered, relative to normal control group (p < 0.05). Four major compounds were identified in the extract: nicotine (45.88 %); 1,5-dimethyl-2-pyrrolidinone (23.00 %), nhexadecanoic acid (11.31 %) and vitamin A aldehyde (9.38 %). Conclusion: These results demonstrate that the methanol extract of smokeless tobacco possesses antidepressant and mood-elevating effects in rats. However, its use should be discouraged since it contains a number of hazardous and carcinogenic components such as N-nitroso compounds and benzo(a)pyrene which are categorized as Class-I carcinogens

Methanol extract of smokeless tobacco alters inflammation and nociception process in animal models

Purpose: To investigate the inflammatory and nociceptive alterations due to the use of Nicotiana tabacum or smokeless tobacco (MEST) owing to the fact that it is used by some people to relieve dental pain.Methods: Hepatic biochemical indicators and thiobarbituric acid reactive substances assay were used to assess MEST toxicity and pharmacological doses selection. The effects on inflammation of different pharmacological doses (100, 200 and 500 mg/kg i.p.) of MEST were evaluated using xylene-induced ear edema and cotton pellet granuloma tests. Indomethacin (10 mg/kg, i.p.) was used as positive standard drug, whereas the vehicle 0.5 % CMC treated group was considered as negative control. Acetic acid-induced abdominal contraction test and formalin-induced hind paw licking model were utilized to assess the role of MEST in nociception. Indomethacin (10 mg/kg i.p.) and diclofenac sodium (10 mg/kg i.p.) were used as positives standard drugs. The vehicle used was 0.5% CMC which served as the negative control.Results: MEST (50 %, 200 mg/kg) and indomethacin (47.5 %) both elicited a significant (p < 0.001) anti-edematogenic effect on xylene-induced ear edema. MEST also showed a significant (p < 0.001) inhibitory effect on granuloma formation at all administered doses as compared to the untreated groups which was comparable to standard drug indomethacin. The number of acetic acid induced writhings was observed to be significantly increased (p < 0.001) by MEST at all doses, unlike diclofenac that led to significant reduction (p < 0.001) in the number of writhings, when compared to the untreated group. MEST also showed a significant (p < 0.05) dose-dependent reduction of the hind paw licking caused by formalin when compared to the vehicle control.Conclusion: These results signify that administration of MEST induces inflammatory and nociceptive alterations. However, the extract is not recommended for dental pain due to its other toxic effects that have previously been reported.Keywords: Nicotiana tabacum, Smokeless tobacco, Inflammation, Nociceptio

Phytochemical profiling of Costus (Saussurea lappa Clarke) root essential oil, and its antimicrobial and toxicological effects

Purpose: To carry out gas chromatography-mass spectrometry (GC-MS) analysis of the phytochemical content of the root essential oil of Saussurea lappa Clarke Asteraceae (Costus, SLEO), and to evaluate its physicochemical, antimicrobial and cytoxic properties. Methods: The oil was extracted from the plant’s roots by steam distillation using a Clevenger system. Various physicochemical parameters for the oil including refractive index, color, acid value, saponification number, ester and peroxide values were measured. Flavonoid content was assessed using thin layer chromatography (TLC). Thermoscientific trace ultra gas chromatograph equipped with a Thermoscientific capillary TR-5MS column was utilized to determine the volatile components of SLEO. Antimicrobial activity of SLEO was performed against various Gram (+ve) and Gram (-ve) microorganisms, viz, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans, while cytotoxic effect was monitored using Artemia salina (brine shrimp) lethality assay. Results: Essential oil yield was good (3 %). Concentration-dependent antimicrobial effects were observed on all test microorganisms and no marked difference in lethality levels was observed among the tested SLEO concentrations on brine shrimp (p < 0.05). The main component of SLEO was costunolide or eudesma-5,11(13)-dien-8,12-olide (52.01 %). Conclusion: The results indicate the promising therapeutic properties of S. lappa. However, further phytochemical and biological investigations are required to establish the mechanism of action and toxicological the extract

High performance thin-layer chromatography and in vitro cytotoxic studies on ethanol extract of Matricaria chamomilla L (Asteraceae) flowers

Purpose: To develop a high performance thin-layer chromatography (HPTLC procedure for quantitation of apigenin in ethanol extract of Matricaria chamomilla (Babunaj) flowers, and to evaluate the extract for in vitro cytotoxic effect on MCF-7 cell lines. Methods: Quantification of apigenin was carried out using a CAMAG TLC system. A combination of toluene, ethyl acetate and formic acid (4.5:3.5:0.2 v/v/v) was used as mobile phase, with densitometry detection at 336 nm. The HPTLC procedure was subjected to validation as per ICH guidelines. The cytotoxicity of the extract was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Results: A sharp apigenin band at Rf of 0.51 was obtained, and the content of apigenin in the extract was 0.062 % w/w. The detection limit (LOD) and quantification limit (LOQ) were 0.19 and 0.57 ng/band, respectively. MTT assay results indicate that M. chamomilla was cytotoxic to Michigan Cancer Foundation-7 (MCF-7) cells, with half-maximal concentration (IC50) of 74 µg/mL. Conclusion: The developed HPTLC method is linear, precise, accurate and specific for the determination of apigenin. M. chamomilla exerts cytotoxic effect on MCF-7 cell line via induction of apoptosis

Phytochemical, antimicrobial and cytotoxicity screening of ethanol extract of Acacia ehrenbergiana Hayne grown in Jazan Region of Saudi Arabia

Purpose: To explore the phytoconstituents of Acacia ehrenbergiana Hayne as well as its biological effects. Methods: Determination of phytoconstituents of ethanol extract of the plant was performed by gas chromatography-mass spectrometry (GC-MS) technique. Antibacterial screening was conducted against the isolates of Gram-positive and Gram-negative microbes while the anti-carcinogenic properties of the ethanol extract on cancerous cells were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cytotoxicity assay against breast MCF7, ovary cancer A2780 and colon cancer HT29 cells, respectively, in addition to normal MRC5 fibroblast cells. Results: GC-MS analysis identified 15 different phytochemicals in the ethanol extract. The extract exerted significant antimicrobial activity with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in the range 1.56 - 6.25 and 3.12 – 12.5 mg/L, respectively, against all test bacterial strains. Cytotoxic activity, obtained by MTT assay, was 28.81 ± 0.99, 12.50 ± 2.50, 23.90 ± 0.74 and 50.58 ± 3.24 μg/mL, against the three cancer cell lines and normal fibroblast, respectively. MTT cytotoxicity results was further confirmed by clonogenic survival assay on MCF7 cells. Conclusion: This study highlights the potential interesting ethnopharmacological applications of Acacia ehrenbergiana Hayne to treat drug-resistant pathogens as standardized extract. Keywords: Acacia ehrenbergiana, Phytochemistry, Antimicrobial, Cytotoxicit

Bioactive principles, antibacterial and anticancer properties of Artemisia arborescens L.

Artemisia arborescens is a medicinal and aromatic plant used in traditionally by the people of Saudi Arabia. This research attempts to evaluate the bioactive constituents of the plant using organic solvents, as well as the antibacterial and anticancer properties of plant extracts. The Phytochemical analysis of methanol extract revealed eleven bioactive constituents, identified by comparing their retention periods and GC-MS profiles to account for 52.45 percent of the studied extract. In the meantime, the extract of pet ether had demonstrated the presence of sixteen significant constituents, six of which were distinct sesquiterpene derivatives. In lipophilic plant extract, three higher alkanes made up 12.49% of the total. These higher alkanes were tetratriacontane (6.55%), hentriacontane (4.17%), and octacosane (1.77%). Studies on antimicrobial activity have revealed that both methanolic and petroleum ether extracts had a broad spectrum of activity against specific human pathogens. Both extracts, however, failed to exhibit any anti-Candida albicans activity. Methanolic extract not shown inhibition in the cell growth of MCF-7 cell, but petroleum ether extract had shown significant anti-cancer activity against MCF-7 cell with an IC50 of 13.49 µg/mL. the results obtained show that A. arborescens have a lot of potential for further research into variety of biological functions, against cancer and microbes.

Mapping an atlas of tissue-specific drosophila melanogaster metabolomes by high resolution mass spectrometry

Metabolomics can provide exciting insights into organismal function, but most work on simple models has focussed on the whole organism metabolome, so missing the contributions of individual tissues. Comprehensive metabolite profiles for ten tissues from adult Drosophila melanogaster were obtained here by two chromatographic methods, a hydrophilic interaction (HILIC) method for polar metabolites and a lipid profiling method also based on HILIC, in combination with an Orbitrap Exactive instrument. Two hundred and forty two polar metabolites were putatively identified in the various tissues, and 251 lipids were observed in positive ion mode and 61 in negative ion mode. Although many metabolites were detected in all tissues, every tissue showed characteristically abundant metabolites which could be rationalised against specific tissue functions. For example, the cuticle contained high levels of glutathione, reflecting a role in oxidative defence; the alimentary canal (like vertebrate gut) had high levels of acylcarnitines for fatty acid metabolism, and the head contained high levels of ether lipids. The male accessory gland uniquely contained decarboxylated S-adenosylmethionine. These data thus both provide valuable insights into tissue function, and a reference baseline, compatible with the FlyAtlas.org transcriptomic resource, for further metabolomic analysis of this important model organism, for example in the modelling of human inborn errors of metabolism, aging or metabolic imbalances such as diabetes