Application of a LC-MS/MS method developed for the determination of p-phenylenediamine, N-acetyl-p-phenylenediamine and N,N-diacetyl-p-phenylenediamine in human urine specimens.

Cases of poisoning by p-phenylenediamine (PPD) are detected sporadically. Recently an article on the development and validation of an LC-MS/MS method for the detection of PPD and its metabolites, N-acetyl-p-phenylenediamine (MAPPD) and N,N-diacetyl-p-phenylenediamine (DAPPD) in blood was published. In the current study this method for detection of these compounds was validated and applied to urine samples. The analytes were extracted from urine samples with methylene chloride and ammonium hydroxide as alkaline medium. Detection was performed by LC-MS/MS using electrospray positive ionization under multiple reaction-monitoring mode. Calibration curves were linear in the range 5-2000 ng/mL for all analytes. Intra- and inter-assay imprecisions were within 1.58-9.52 and 5.43-9.45%, respectively, for PPD, MAPPD and DAPPD. Inter-assay accuracies were within -7.43 and 7.36 for all compounds. The lower limit of quantification was 5 ng/mL for all analytes. The method, which complies with the validation criteria, was successfully applied to the analysis of PPD, MAPPD and DAPPD in human urine samples collected from clinical and postmortem cases.

Potential of Ficus microcarpa metabolites against SARS-CoV-2 main protease supported by docking studies.

COVID 19; an infectious disease; firstly identified in December 2019 in Wuhan, China and has since spread globally, resulting in an ongoing pandemic. Searching for protease inhibitors is a challenging task in controlling COVID 19. Genus Ficus is known to be a rich source of phenolic compounds. Metabolic profiling of leaves methanolic extract of Ficus microcarpa (Moraceae) revealed nine compounds (1-9) mainly phenolics. Docking studies concerning these compounds against SARS-CoV-2 main protease showed that quercetin 3,7-O-α-L-dirhamnoside (1) and rutin (3) possessed significant binding stability at the N3 binding site in different activity degrees, which is comparable with COVID-19 main protease inhibitor, darunavir. Our study suggests that compounds quercetin 3,7-O-α-L-dirhamnoside and rutin might be potential candidates for the development of therapies against SARS-CoV-2.

Isolation and characterization of novel acetylcholinesterase inhibitors from Ficus benghalensis L. leaves.

Metabolic profiling of the crude methanolic extract of Ficus benghalensis leaves has revealed the presence of different phenolic and nitrogenous compounds including cerebrosides and tetrapyrrole pigments. A phytochemical study of the ethyl acetate fraction resulted in the identification of three known compounds, namely carpachromene (1), alpha amyrine acetate (2), and mucusoside (3) together with one new fatty acid glycoside, named 2-O-α-l-rhamnopyranosyl-hexacosanoate-ß-d-glucopyranosyl ester (4). The compounds were identified using 1D, 2D NMR, and HR-ESIMS techniques as well as via comparison to other literature. Studies on the acetylcholinesterase inhibition potential and antioxidant activity were carried out on the total methanolic leaf extract, ethyl acetate fraction, and the isolated compounds. The results revealed the potent acetylcholinesterase inhibition of mucusoside alongside a new compound. Docking studies were also performed to confirm the possible interaction between the isolated compounds and acetylcholinesterase accompanying Alzheimer's disease progress.

GC-MS Method for Quantification of Mephedrone in Human Blood Sample.

Abuse of mephedrone (4-methylmethcathinone) has emerged among the last few years. Intoxication and fatality associated with mephedrone have been reported in the literature. The present work describes a simple and rapid gas chromatography-mass spectrometry (GC-MS) method for detection and quantification of mephedrone in human blood samples using GC-MS. Blood was deproteinated with acetonitrile and the supernatant was derivatized and extracted in a single step with ethyl chloroformate and ethyl acetate. Methamphetamine-d5 was used as an internal standard (IS). The characteristic ions at m/z 58, 119 and 130 for mephedrone and m/z 62, 92 and 134 for methamphetamine-D5 were used for quantification of the analyte. The calibration curve was linear within the concentration range 10-2,000 ng/mL. Intra- and inter-assay precisions (%RSD) were within 10.9-11.9 and 9.2-11.2, respectively. Intra-assay accuracy (%Bias) was between 1.8 and 2.8. The method was successfully applied to detect and quantify mephedrone in real blood samples.

A practical application of photogrammetry to performing rib characterization measurements in an underground coal mine using a DSLR camera.

Understanding coal mine rib behavior is important for inferring pillar loading conditions as well as ensuring the safety of miners who are regularly exposed to ribs. Due to the variability in the geometry of underground openings and ground behavior, point measurements often fail to capture the true movement of mine workings. Photogrammetry is a potentially fast, cheap, and precise supplemental measurement tool in comparison to extensometers, tape measures, or laser range meters, but its application in underground coal has been limited. The practical use of photogrammetry was tested at the Safety Research Coal Mine, National Institute for Occupational Safety and Health (NIOSH). A commercially available, digital single-lens reflex (DSLR) camera was used to perform the photogrammetric surveys for the experiment. Several experiments were performed using different lighting conditions, distances to subject, camera settings, and photograph overlaps, with results summarized as follows: the lighting method was found to be insignificant if the scene was appropriately illuminated. It was found that the distance to the subject has a minimal impact on result accuracy, and that camera settings have a significant impact on the photogrammetric quality of images. An increasing photograph resolution was preferable when measuring plane orientations; otherwise a high point cloud density would likely be excessive. Focal ratio (F-stop) changes affect the depth of field and image quality in situations where multiple angles are necessary to survey cleat orientations. Photograph overlap is very important to proper three-dimensional reconstruction, and at least 60% overlap between photograph pairs is ideal to avoid unnecessary post-processing. The suggestions and guidelines proposed are designed to increase the quality of photogrammetry inputs and outputs as well as minimize processing time, and serve as a starting point for an underground coal photogrammetry study.

Comparison of 3 Derivatization Methods for the Analysis of Amphetamine-Related Drugs in Oral Fluid by Gas Chromatography-Mass Spectrometry.

The heptafluorobutyric anhydride (HFBA), pentafluoropropionic anhydride (PFPA), and trifluoroacetic anhydride (TFAA) are compared as derivatizing reagents to use as the optimal method for the analysis of 10 amphetamines and cathinones in oral fluid. The target compounds were amphetamine (AMP), methamphetamine (MA), 4-methylamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), cathinone (CAT), methcathinone, mephedrone, and ephedrine. Amphetamine-D5, MA-D5, MDA-D5, MDMA-D5, and MDEA-D5 use as internal standards (IS). The analytes and IS were extracted from 0.5 mL of oral fluid by ethyl acetate in the presence of NaOH (0.1 N) as the base and then the dried extracts were derivatized with HFBA, PFPA, or TFAA at 70°C for 30 minutes. The limits of quantification based on signal-to-noise ratios ≥10 were ranged between 2.5 and 10 ng/mL. The calibration graphs were linear in the range of 5 or 10 to 1000 ng/mL for all analytes. Based on sensitivity, the PFPA is proved to be the best for derivatization of the target compounds prior to gas chromatography-mass spectrometry analysis.

Use of Herbal Medicines in the Treatment of Obesity in Taif, Saudi Arabia.

BACKGROUND: The prevalence of obesity is increasing worldwide. Herbal medicines are one of the most common alternative therapies for weight loss. The aim of this study was to determine the usage rate of herbal medicines in the city of Taif, Saudi Arabia, and which plants were most commonly used to treat obesity. MATERIALS AND METHODS: This is a cross-sectional study conducted using a pretested questionnaire that was randomly distributed to overweight and obese people in Taif, Saudi Arabia. RESULTS: A total of 422 cases completed the questionnaire with a response rate of 60.3%. The majority of patients (98.1%) used herbal medicines to lose weight, with green tea (88.4%) and ginger (29.5%) being the most commonly used herbs. More than one-third of respondents declared that they suffered from side effects; however, most (72%) of the respondents admitted that they would use herbal medications again. CONCLUSION: The current study shows a high usage rate of herbal medicines for the treatment of obesity in Saudi Arabia. However, further studies are needed on the risks and benefits of the use of herbal medicines to treat obesity. Pharmacists and health professionals should conduct a community-based awareness program to spread awareness of both the benefits and risks of medicinal herbs.

Phenylpropanoid and phenylethanoid derivatives from Kigelia pinnata DC. fruits.

Further phytochemical investigation of the fruits of Kigelia pinnata DC. has yielded a new phenylpropanoid derivative identified as 6-p-coumaroyl-sucrose (1) together with ten known phenylpropanoid and phenylethanoid derivatives (2-11) and a flavonoid glycoside (12). The structures of the isolated compounds were elucidated using various techniques of NMR and MS spectral analysis.

Analysis of the current rib support practices and techniques in U.S. coal mines.

Design of rib support systems in U.S. coal mines is based primarily on local practices and experience. A better understanding of current rib support practices in U.S. coal mines is crucial for developing a sound engineering rib support design tool. The objective of this paper is to analyze the current practices of rib control in U.S. coal mines. Twenty underground coal mines were studied representing various coal basins, coal seams, geology, loading conditions, and rib control strategies. The key findings are: (1) any rib design guideline or tool should take into account external rib support as well as internal bolting; (2) rib bolts on their own cannot contain rib spall, especially in soft ribs subjected to significant load-external rib control devices such as mesh are required in such cases to contain rib sloughing; (3) the majority of the studied mines follow the overburden depth and entry height thresholds recommended by the Program Information Bulletin 11-29 issued by the Mine Safety and Health Administration; (4) potential rib instability occurred when certain geological features prevailed-these include draw slate and/or bone coal near the rib/roof line, claystone partings, and soft coal bench overlain by rock strata; (5) 47% of the studied rib spall was classified as blocky-this could indicate a high potential of rib hazards; and (6) rib injury rates of the studied mines for the last three years emphasize the need for more rib control management for mines operating at overburden depths between 152.4 m and 304.8 m.

A GC-MS Method for Detection and Quantification of Cathine, Cathinone, Methcathinone and Ephedrine in Oral Fluid.

The stimulating herbal drug khat (cathine, cathinone) and its analog methcathinone are common substances of abuse in most countries. A GC-MS method was developed and validated for the detection and quantification of cathine, cathinone, methcathinone and ephedrine in oral fluid specimens. The analytes and internal standard (amphetamine-d5) were extracted from 0.5 mL oral fluids by ethyl acetate, and then the dried extracts were derivatized with heptafluorobutyric anhydride at 70°C for 30 min. The MS was used in selected ion monitoring mode. Ions monitored were m/z 117, 240 and 330 for cathine, m/z 77, 105 and 240 for cathinone, m/z 105, 210 and 254 for methcathinone, m/z 210, 254 and 344 for ephedrine and m/z 244 and 336 for amphetamine-d5 The calibration curves were linear (r(2)> 0.98) in the concentration range 20-2,000 ng/mL for all analytes. The intra- and inter-assay imprecisions were within (1.6-12.5%) and (1.5-9.5%), respectively, for all analytes. Intra-assay accuracies were between -5.9 and 6.7% for all analytes. The method was successfully applied to detect and quantify the target analytes from oral fluid specimens collected from Khat and methcathinone users.

Two new cytotoxic furoquinoline alkaloids isolated from Aegle marmelos (Linn.) Correa.

Two new cytotoxic furoquinoline alkaloids were isolated from the leaves of Aegle marmelos (Linn.) Correa; one from the total alkaloidal fraction (acid/base shake-out method) of the CHCl3 extract and identified as 7,8-dihydroxy-4-hydrofuroquinoline and named trivially as Aegelbine-A. The other new alkaloid isolated from the pet. ether extract and identified as 4-hydro-7-hydroxy-8-prenyloxyfuroquinoline and named trivially as Aegelbine-B, together with a known alkaloid; aegeline and a known phenolic acid; ρ-hydroxybenzoic acid. The structures of all the isolated compounds were established based on 1D and 2D NMR spectroscopy and HR-ESI/MS. The cytotoxic activity of the isolated compounds was evaluated in vitro against HepG-2, PC3, A549 and MCF-7 cell lines. The obtained results revealed promising activity with structure-based relationship which is discussed briefly.

Development and validation of an LC-MS/MS method for determination of p-phenylenediamine and its metabolites in blood samples.

In some developing countries, p-phenylenediamine (PPD) is used in combination with Henna as hair dye or skin decoration. A sensitive LC-MS/MS method was developed and validated for the simultaneous determination of p-phenylenediamine (PPD) and its metabolites N-acetyl-p-phenylenediamine (MAPPD) and N,N-diacetyl-p-phenylenediamine (DAPPD) in human blood. Acetanilide was used as an internal standard (IS). The LC-MS/MS was operated under multiple reaction-monitoring mode using the electrospray positive ionization technique. The transition ions m/z 109→92, m/z 151→92, m/z 193→92, and m/z 136→77 were selected for the quantification of PPD, MAPPD, DAPPD, and IS, respectively. The linear range was 10-2000ng/mL for all the compounds. The absolute recoveries were 51.94, 56.20 and 54.88% for PPD, MAPPD and DAPPD, respectively. Intra- and inter-assay imprecision were lower than 14% (RSD), and the bias of the assay was lower than 15% for all the compounds. The stability studies demonstrated that critical degradation for PPD in blood samples and autosampler occurred after 6h, while MAPPD and DAPPD were stable in blood samples and the autosampler up to 48h and 24h, respectively. This newly developed method allows for the detection of PPD and its metabolites in blood samples in the clinical and forensic setting.

Effects of fenugreek, Nigella, and termis seeds in nonalcoholic fatty liver in obese diabetic albino rats.

BACKGROUND AND STUDY AIM: Nonalcoholic fatty liver disease (NAFLD) occurs in approximately 80% of cases of type 2 diabetes mellitus (T2DM). This study investigated the effects of some plants used in Saudi Arabia as antidiabetic agents on T2DM and associated fatty liver. MATERIALS AND METHODS: A total of 150 adult male albino rats were divided into six experimental groups, each consisting of 25 rats. Twenty-five rats were considered as the control group. Experimental diabetes was induced in the remaining rats by administering a subcutaneous injection of 120 mg/kg of freshly prepared alloxan solution; these rats were classified into five groups: one group did not receive any treatment; the second group was treated with an aqueous extract of a mixture containing fenugreek, Nigella, and termis seeds; the third group was treated with an aqueous extract of Nigella sativa seeds; the fourth group was treated with an aqueous extract of fenugreek seeds; and finally the fifth group was treated with an aqueous extract of termis seeds at a dose of 100 mg/kg body weight. After 4 weeks of treatment, biochemical parameters were calculated, including blood sugar and serum insulin levels. Pancreatic and liver samples were obtained and processed for microscopic evaluation. RESULTS: The usage of each plant alone or a mixture of the plants corrected the glucose and insulin levels. Microscopically, a definite improvement in the number and diameter of ß-cells in the diabetic group was observed. Furthermore, considerable improvement in fatty changes occurring in the liver of experimental animals was observed. The activity of the mixture was the most effective. CONCLUSIONS: The aqueous extract of the seed mixture of the used plants appeared to be a useful agent in improving fatty changes in the liver texture associated with T2DM by reducing hyperglycaemia through an increase in insulin levels, regeneration of ß-cells of the pancreas, and an amelioration of associated dyslipidemia.

Iridoids from Kigelia pinnata DC. fruits.

From the fruits of Kigelia pinnata DC., a new furanone derivative formulated as 3-(2'-hydroxyethyl)-5-(2"-hydroxypropyl)-dihydrofuran-2(3H)-one (1), and four new iridoids named; 7-hydroxy viteoid II (2), 7-hydroxy eucommic acid (3), 7-hydroxy-10-deoxyeucommiol (4) and 10-deoxyeucommiol (5) have been isolated together with seven known iridoids, jiofuran (6), jioglutolide (7), 1-dehydroxy-3,4-dihydroaucubigenin (8), des-p-hydroxybenzoyl kisasagenol B (9), ajugol (10), verminoside (11) and 6-trans-caffeoyl ajugol (12). The structures of the isolated compounds were characterized by different spectroscopic methods.

Simultaneous determination of tramadol and O-desmethyltramadol in human plasma using HPLC-DAD.

A sensitive and accurate method for the extraction and quantification of tramadol (T) and its active metabolite, O-desmethyltramadol (ODT) in human plasma with high-performance liquid chromatography-diode array detection was developed and validated. The analytes were extracted from plasma samples by tert-butylmethyl ether in the presence of ammonium hydroxide as alkaline medium and back extraction with 1.0 M hydrochloric acid. Propranolol was used as internal standard. The extraction efficiencies of T and ODT were 83.51 and 78.72%, respectively. The calibration curves were linear (r(2) > 0.99) in the concentration range of 250-2000 ng/mL for T and ODT. Limits of detection and quantification were 125 and 250 ng/mL for both analytes. Intra- and interassay precision for T and ODT were ranged from 1.89 to 10.91% and 2.16 to 5.85%, respectively. Intra- and interassay accuracy for T and ODT were ranged from -13.07 to 4.99% and -2.03 to -6.98%, respectively. The method was successfully applied to quantify T and ODT from authentic plasma samples received from Hospital Sohag University. The method was completely validated and can be of interest to clinical and forensic laboratories.

Simultaneous determination of tramadol, O-desmethyltramadol and N-desmethyltramadol in human urine by gas chromatography-mass spectrometry.

Analytical procedures for the determination of tramadol (T), O-desmethyltramadol (ODT), and N-desmethyltramadol (NDT) in human urine have been developed and validated using gas chromatography-mass spectrometry (GC/MS). Sample preparation involved liquid-liquid extraction with methyl-tert-butyl ether (MTBE) and followed by back extraction with 0.1 M hydrochloric acid. Proadifen (SKF525A) was selected as internal standard (IS). Extraction efficiencies of T, ODT and NDT were 102.12, 101.30, and 98.21%, respectively. The calibration curves were linear (r(2)>0.99) in the concentration range 10-1000 ng/mL for all compounds. Limits of quantification (LOQ) were 10, 10 and 20 ng/mL for T, ODT and NDT, respectively. Intra-assay precision was within 1.29-6.48% and inter-assay precision was within 1.28-6.84% for T, ODT and NDT. Intra-assay accuracy was within 91.79-106.89% for all analytes. This method detected urine concentrations of T, ODT and NDT in six healthy volunteers for 7 days after administration of 50 mg oral doses of tramadol.

Nonclassical antifolates, part 4. 5-(2-aminothiazol-4-yl)-4-phenyl-4H-1,2,4-triazole-3-thiols as a new class of DHFR inhibitors: synthesis, biological evaluation and molecular modeling study.

A new series of compounds possessing 5-(2-aminothiazol-4-yl)-4-phenyl-4H-1,2,4-triazole-3-thiol skeleton was designed, synthesized, and evaluated for their in vitro DHFR inhibition, antimicrobial, antitumor and schistosomicidal activities. Four active compounds were allocated, the antibacterial 22 (comparable to gentamicin and ciprofloxacin), the schistosomicidal 29 (comparable to praziquantel), the DHFR inhibitor 34 (IC50 0.03 µM, 2.7 fold more active than MTX), and the antitumor 36 (comparable to doxorubicin). Molecular modeling studies concluded that recognition with key amino acid Leu4 and Val1 is essential for DHFR binding. Flexible alignment and surface mapping revealed that the obtained model could be useful for the development of new class of DHFR inhibitors.

Induction of Biologically Active Flavonoids in Cell Cultures of Morus nigra and Testing their Hypoglycemic Efficacy.

The antidiabetic activity of both leaves and MJ-treated cell cultures of Morus nigra was evaluated after their oral administration to streptozotocin-induced diabetic rats. The antidiabetic activity of extracts from leaves given to streptozotocin (STZ)-diabetic rats for 10 days increased with increasing doses of leaves extract up to 500 mg/kg/day. The administration of 500 mg/kg/day of leaves extract reduced the concentration of glucose from 370 ± 7.31 mg/dl (control) to 154 ± 6.27 mg/dl, and a significant increase in the insulin level from 11.3 ± 0.31 µU/ml (control) to 14.6 ± 0.43 µU/ml was recorded. Cell suspension cultures were established from the young leaves of Morus nigra cultivated on modified MS medium supplemented with 2.0 mg/l 1-naphthaleneacetic acid (NAA), 0.2 mg/l 6-(furfurylamino)purine (kinetin). The changes in cell weight and flavonoid content were monitored between day zero and 12. The linear increase in fresh weight was found to be parallel to flavonoids production. Cell cultures treated with 100 µM methyl jasmonate for 24 hours showed a noticeable increase in level of flavonoids and significant and more effective hypoglycemic activity than that for extract from leaves. The major flavonoids were isolated by TLC and HPLC and identified as rutin, quercetin, Morusin and cyclomorusin by co-chromatography and mass spectrometry in comparison to samples of authentic reference compounds.

Cesdiurins I-III, steroidal saponins from Cestrum diurnum L.

Phytochemical investigation of a methanolic extract of leaves of Cestrum diurnum L (Solanaceae) resulted in isolation of several furostanol steroidal saponins, named cesdiurins I-III (1-3). Their structures of the isolated compounds were determined by spectroscopic analyses, including by use of 1D and 2D NMR spectroscopic techniques as well as by mass spectral analyses.

Zoonotic cutaneous leishmaniasis: reservoir host and insect vector in north Sinai, Egypt.

Cutaneous leishmaniasis (ZCL) is a protozoan disease well documented not only in Egypt, but in nearly all the East Mediterranean Countries. Sinai Peninsula was a sparsely populated area where sporadic cases of ZCL were reported with the reconstruction of Sinai and people coming in and out, it was indicated to study the status of ZCL in North Sinai Governorate, the reservoir host(s) and insect vector(s) in Sinai. In the present study, the six species of rodents were trapped from areas or nearby areas where human ZCL cases were detected. Rodents (50) were Mus musculus (10), Rattus r. alexandrinus (18), R. norvegicus (2), Gerbillus gerbillus (4), G. pyramidum (12) and Jaculus jaculus (4). The rodents were examined clinically for any skin lesion or even nodule, particularly in head and tail. One G. pyramidumn had natural infection with L. major as indicated by smears and culture, but typing was not done. The spot light surveys for Phlebotomus were carried out by the sticky paper traps and the CDC light traps in four main centers; Al Hassanah, Nakhil, Al Arish, and Bir Al-Abd. A total of 1320 sandflies were identified. They were P. papatasi (1150) and P. sergenti (170) in a ratio of 7:1. A total of three isolates of zymodeme London 70 undistinguished from the formerly obtained human and rodent isolates were enzymatically identified in P. papatasi.