Effect of olive leaf phytochemicals on the anti-acetylcholinesterase, anti-cyclooxygenase-2 and ferric reducing antioxidant capacity.

In this study, the phytochemical profile of fifty olive leaves (OL) extracts from Spain, Italy, Greece, Portugal, and Morocco was characterized and their anti-cholinergic, anti-inflammatory, and antioxidant activities were evaluated. Luteolin-7-O-glucoside, isoharmnentin, and apigenin were involved in the acetylcholinesterase (AChE) inhibitory activity, while oleuropein and hydroxytyrosol showed noteworthy potential. Secoiridoids contributed to the cyclooxygenase-2 inhibitory activity and antioxidant capacity. Compounds such as oleuropein, ligstroside and luteolin-7-O-glucoside, may exert an important role in the ferric reducing antioxidant capacity. It should be also highlighted the role of hydroxytyrosol, hydroxycoumarins, and verbascoside concerning the antioxidant activity. This research provides valuable insights and confirms that specific compounds within OL extracts contribute to distinct anti-cholinergic, anti-inflammatory, and anti-oxidative effects.

In vitro anticholinergic and antiglycaemic properties of frost-hardy Actinidia fruit extracts and their polyphenol profile, L-ascorbic acid content and antioxidant capacity.

The aim of this study was to investigate the inhibitory effects of Actinidia arguta ('Weiki', 'Skarlet September Kiwi') and Actinidia kolomikta ('Lande') fruit extracts against advanced glycation end-products (AGEs) formation and acetylcholinesterase (AChE) activity. The extracts were also tested regarding polyphenol profile and Lascorbic acid content (UHPLC-DAD-MS), and antioxidant capacity (DPPH, ABTS). 'Scarlet September Kiwi' showed the strongest anti-AGEs activity studied with BSAGLU (IC50 = 2.68) and BSA-MGO (IC50 = 18.06) models. The highest anti-AChE activity was found for the 'Lande' extract (IC50 = 4.56). 'Lande' showed the highest L-ascorbic acid content (8271.96 µg/g dw), ABTS (312.42 µmol TE/g dw) and DPPH (282.01 µmol TE/g dw) values. 'Scarlet September Kiwi' revealed the highest individual phenolics concentration (2321.43 µg/g dw). The contents of (+)-catechin and L-ascorbic acid were significantly correlated with anti-AChE activity. This research sheds new light on the bioactivity of Actinidia arguta and Actinidia kolomikta fruit elucidating the role of (+)-catechin and L-ascorbic acid in prevention of Alzheimer's disease.

Variables Affecting Delivery of Glycopyrronium Tosylate Through Human Skin In Vitro.

BACKGROUND: Hyperhidrosis is a condition characterized by excessive sweating beyond what is required for normal thermal regulation. It can involve multiple body areas including the axillae, palms, soles, or craniofacial regions. Glycopyrronium tosylate (GT) is a topical anticholinergic approved by the FDA (2018) for treatment of primary axillary hyperhidrosis in patients 9 years and older. OBJECTIVE: Gain insight into variables (anatomical sites, occlusion, exposure time) affecting GT delivery into human skin. METHODS: Human skin from different anatomical regions (palmar, plantar, axillary, and abdominal skin) was mounted into flow-through diffusion cells (MedFlux-HT®). GT solution (2.4%) was applied at 10 mg/cm2 and the receiving fluid was collected every 2 hours, for 24 hours. GT penetration was determined using LC/MS/MS. The effect of occlusion was assessed by covering the skin with either parafilm or saran wrap, and the effect of exposure time was assessed by incubating the skin for 5, 15, or 60 minutes before washing off the GT from the surface. RESULTS: GT delivery through palmar and plantar skin was up to 40-fold lower compared to delivery through axillary or abdominal skin. Occlusion increased GT delivery up to 10-fold. Reducing exposure time from 24 hours to either 5, 15, or 60 minutes, decreased GT flux by 90%. However, occlusion during these varied exposure times was able to restore GT delivery to levels found in the 24-hour exposed, non-occluded control group. CONCLUSION: These in vitro skin penetration studies showed that skin thickness, exposure time, and occlusion substantially influenced GT delivery, potentially impacting clinical trial design. J Drugs Dermatol. 2020;19(11): doi:10.36849/JDD.2020.5062.

Simple validated method of LC-MS/MS determination of BZ agent in rat plasma samples.

Agent BZ (3-quinuclidinyl benzilate) is a centrally acting synthetic anticholinergic agent, considered as a potential military incapacitating chemical warfare agent. Despite its significance as a model compound in pharmacological research and its potential misuse in chemical attacks, few modern analytical methods for BZ determination in biological samples have been published. The goal of the present work is to develop and validate a sensitive and rapid LC-MS/MS method for the determination of agent BZ in rat plasma. The sample preparation was based on solid-phase extraction on C-18 cartridges. The reversed-phase HPLC coupled with the mass spectrometer with electrospray ionization in the positive ion-selective reaction monitoring mode was employed in the BZ analysis. Atropine was used as an internal standard. The presented method is selective, accurate, precise, and linear (r2 = 0.9947) in a concentration range from 0.5 ng/mL to 1 000 ng/mL and sensitive enough (limit of detection 0.2 ng/mL; limit of quantification 0.5 ng/mL) to determine the BZ plasma levels in rats exposed to 2 mg/kg and 10 mg/kg of BZ. The highest level of BZ in plasma was observed 5 minutes after intramuscular administration (154.6 ± 22.3 ng/mL in rats exposed to 2 mg/kg of BZ and 1024 ± 269 ng/mL in rats exposed to 10 mg/kg). After 48 h, no BZ was observed at detectable levels. This new method allows the detection and quantification of BZ in biological samples after exposure of an observed organism and it will be further optimized for other tissues to observe the distribution of BZ in organs.

Preliminary phytochemical analysis and evaluation of in vitro antioxidant, antiproliferative, antidiabetic, and anticholinergics effects of endemic Gypsophila taxa from Turkey.

The phenolic contents and antioxidant, anticancer, antidiabetic, and anticholinergic potentials of four endemic Gysophila taxa (G. pallida, G. arrosti, G. tuberculosa, and G. eriocalyx) were investigated. The HPLC analysis showed that methanol extracts of all the tested species were richer in phenolics than water extracts. 3,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, vanillin, syringic acid, and p-coumaric acid were detected in all extracts. In parallel to the phenolic contents, methanol extracts displayed comparatively higher antioxidant activity than water extracts. Additionally, all extracts exhibited dose-dependent antiproliferative activity on the cancer cell lines with lower IC50 values changing from 0.170 to 1.805 mg/ml. Moreover, the extracts impressively inhibited the acetylcholinesterase (0.63-26.04), butyrylcholinesterase (3.66-10.73), and α-glycosidase (98.52-235.55) enzymes with very low IC50 (mg/ml) values. Together, the present results indicate that Gysophila taxa have various biological activities together with higher phenolic contents. Hence, these species hold good potential for use in the pharmaceutical industry. PRACTICAL APPLICATIONS: Gypsophila taxa having numerous biological activities have been used for different purpose in folk medicine as well as their use in the food industry. The obtained results of the current study indicated that the extracts of Gypsophila taxa are rich in phenolics and flavonoids with powerful antioxidant and antiproliferative activity against different type of cancer cell lines. In addition, the extracts obtained from these taxa showed notable antidiabetic and anticholinergics effects. Gypsophila taxa could be used as a natural material to develop anticancer, antidiabetic, and anticholinergic drugs.

Anticholinergic effects of Actinidia arguta fruits and their polyphenol content determined by liquid chromatography-photodiode array detector-quadrupole/time of flight-mass spectrometry (LC-MS-PDA-Q/TOF).

This study discusses polyphenolic compounds identified and quantified in Actinidia arguta fruits by LC-MS-PDA-Q/TOF method and in vitro anticholinergic activity. Notably, of 31 compounds, including 16 flavonols, 7 flavanols, 7 phenolic acids, and 1 anthocyanin were identified or tentatively identified on the basis of their retention times, accurate mass measurements and subsequent mass fragmentation data, or by comparison with reference substances and literature. Among the detected compounds, 27 were reported for the first time in A. arguta fruits. The content of total polyphenols equal 845.54 mg/100 g dry weight (dw), and flavanols predominat (92% of total phenolic compounds). Flavonol derivatives, mainly glycosylated and acetylated forms of quercetin (22.64 mg/100 g dw) and kaempferol (18.40 mg/100 g dw) were quantified. The total content of phenolic acids was 29.63 mg/100 g dw, and neochlorogenic acid predominant. This anticholinergic activity effect of A. arguta fruits can be explained by the Pearson's correlation found between flavonols (r = 0.709 and 0.678), phenolic acids (r = 0.513 and 0.487), flavan-3-ols (r = 0.466 and 0.443) and anthocyanins (r = 0.312 and 0.301) for acetylcholinesterase (AChE) or butylcholinoesterase (BuChE), respectively. The data compiled from the quantitative polyphenol indicate that A. arguta fruits could be regarded as a promising source of bioactive functional food.

[Anticholinergic syndrome caused by contaminated herbal tea; acting swiftly to identify the source]. / Anticholinerg syndroom door verontreinigde kruidenthee.

BACKGROUND: Despite good manufacturing practice and quality control, consumer products can become contaminated. In some cases, this can result in severe and life-threatening intoxication with potentially fatal consequences. CASE DESCRIPTION: A 27-year-old man and a 28-year-old pregnant woman presented to the Emergency Department with severe anticholinergic syndrome after using a marshmallow root (Althaea officinalis) herbal remedy, mixed into hot chocolate drink, to reduce symptoms of common cold. After a short stay in Intensive Care, the symptoms diminished and the patients could be released from hospital. The herbs were found to be contaminated with atropine, most probably derived from deadly nightshade (Atropa belladonna). Analyses of the contaminated product indicated that the patients were exposed to 20-200 mg atropine, while a dose of 2 mg is already considered mildly toxic. CONCLUSION: Consultation of the Dutch National Poisons Information Center resulted in rapid detection of the contamination; close collaboration with the Netherlands Food and Consumer Product Safety Authority and the manufacturer of the product allowed rapid identification of the source of contamination and facilitated the prevention of an epidemic.

Toxicological results in a fatal and two non-fatal cases of scopolamine-facilitated robberies.

The use of scopolamine as an incapacitating drug, in sexual crimes and robberies, has been known for many decades. However, blood concentrations and doses of scopolamine in those cases are largely unknown. Here we present the toxicological results of one fatal and two non-fatal cases in a series of scopolamine-facilitated robberies. In the fatal case, the concentration of scopolamine in heart blood was 0.30mg/L, about 3000 times higher than the average therapeutic level of 0.0001mg/L (for one dermal patch). In femoral blood, the concentration of scopolamine was much lower (0.0048mg/L), but still 50 times higher than therapeutic levels. The scopolamine concentration in the stomach was very high (20mg/kg) as compared to the heart blood and femoral blood, which explains the very high concentration in heart blood by postmortem leakage from the stomach. In the non-fatal case, the scopolamine concentration in serum, obtained 23h after the incident, was 0.00035mg/L. The estimated concentration of scopolamine at the time of the incident is 0.0035mg/L. In the other non-fatal case, scopolamine was detected in urine and in hair.

Cholinesterase-based biosensors.

Recently, cholinesterase-based biosensors are widely used for assaying anticholinergic compounds. Primarily biosensors based on enzyme inhibition are useful analytical tools for fast screening of inhibitors, such as organophosphates and carbamates. The present review is aimed at compilation of the most important facts about cholinesterase based biosensors, types of physico-chemical transduction, immobilization strategies and practical applications.

Acute benztropine intoxication and fatality.

A woman was found unresponsive with an empty bottle of Cogentin(®) prescribed to another. Admitted to an area hospital, her condition steadily declined until death 29 h after admission. Following toxicological screening on hospital (admission) whole blood, the only significant compound detected was benztropine. Benztropine was confirmed at 0.28 mg/L - the highest antemortem blood concentration recorded in a case of toxicity or fatality uniquely associated with benztropine. A second serum antemortem specimen showed a benztropine concentration of 0.19 mg/L. Despite over 24 h in the hospital, benztropine was also found in the postmortem specimens collected at autopsy. Peripheral blood, central blood, liver, and gastric concentrations were 0.47 mg/L, 0.36 mg/L, 9.6 mg/kg, and 44 mg, respectively. These results indicate that benztropine exhibited a potential difference between whole-blood and serum (plasma) concentrations. Additionally, in consideration of literature data, benztropine was found indicative of a compound prone to at least some postmortem redistribution.

Fast determination of two atractylenolides in Rhizoma Atractylodis Macrocephalae by Fourier transform near-infrared spectroscopy with partial least squares.

Rhizoma Atractylodis Macrocephalae (RAM) is a commonly used food and traditional Chinese medicine (TCM), which traditionally strengthens the spleen, benefits vital energy, eliminates dampness, and promotes hidroschesis. Its primary effective constituents are polysaccharides and volatile oil, whose main components are atractylenolide I and III. Fourier transform near-infrared spectroscopy (FT-NIR) is widely used in TCM research. However, determination of atractylenolides in RAM using FT-NIR has not been described. In this study, a new method for the determination of atractylenolides I and III in RAM by NIR was established. The spectral characteristics of atractylenolides I and III were obtained by second derivative multiple scattering correction, and its chart to the original absorbance spectra. Additionally, in combination with the partial least squares (PLS) algorithm, the calibration process was performed for the quantitation of the samples. The root mean square error of cross-validation of the PLS models for atractylenolides I and III was 0.0387 and 0.0358, and the determination coefficient of quantitative models was 96.63 and 96.16, respectively. This study demonstrated that NIR spectroscopy can be used to analyze quickly and efficiently the contents of atractylenolides I and III in RAM.

Determination of bencycloquidium bromide, a novel anticholinergic compound, in rat tissues by liquid chromatography-electrospray ionization mass spectrometry.

First, a liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method for quantification of bencycloquidium bromide (BCQB) in rat tissue homogenates was developed and validated, which would support investigation on drug distribution into tissues in animal models. 1-ethyl-bencycloquidium bromide was used as the internal standard (IS). Sample preparation in tissue homogenates was achieved by using solid phase extraction on a 3 mL C(18)-cartridge column. Chromatographic separation was analyzed on a Hanbon Lichrospher 5-C(18) column. The mobile phase consisted of methanol-40 mM ammonium acetate buffer-formic acid (75:25:0.25, v/v/v) which was pumped at 1.0 mL min(-1). BCQB was determined using electrospray ionization in a single quadrupole mass spectrometer. LC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 330.2 for BCQB and m/z 344.2 for the IS. The assay was linear from 3.015 ng mL(-1) to 301.5 ng mL(-1) of BCQB in rat tissue (liver, kidney, lung, trachea, heart, spleen, stomach, intestines, brain, muscle, testicle, ovary and fat) homogenates. The lower limit of quantification (LLOQ) was 3.015 ng mL(-1) of BCQB in all tissue homogenates. Acceptable precision and accuracy were obtained for concentrations over the entire standard curve ranges for tissue homogenates. The method was used to successfully quantify BCQB in rat tissue homogenates for a tissue distribution study of BCQB in rats after intranasal administration.

Electrogenerated chemiluminescence of ruthenium complex immobilized in a highly cross-linked polymer and its analytical applications.

Electrogenerated chemiluminescence (ECL) of a ruthenium complex polymer modified carbon paste electrode and its analytical applications were investigated. The ruthenium complex polymer was prepared using bis(2,2-bipyridine) (4,4-dicarboxy-2,2-bipyridine) ruthenium(II). The ECL behaviours of ruthenium complex polymer modified carbon paste electrode were investigated in the absence and presence of tripropylamine (TPA). The modified carbon paste electrode exhibited long-term stability and fine reproducibility. The ECL intensity of the modified carbon paste electrode was linear with the concentration of TPA in the range 2.0 x 10(-6)-3.8 x 10(-3) mol/L, with a detection limit (S:N = 3) of 6 x 10(-7) mol/L. It was also found that raceanisodamine could enhance the ECL intensity of the modified electrode. The ECL intensity of the modified carbon paste electrode was linear with the concentration of raceanisodamine in the range 1.1 x 10(-5)-6.0 x 10(-4) mol/L, with a detection limit (S:N = 3) of 6 x 10(-6) mol/L. This work demonstrates that the entrapment of ruthenium complex in a highly cross-linked polymer is a promising approach to construct an ECL modified electrode with long-term stability and fine reproducibility. The modified electrode designed has a potential application in the ECL detector.

Method development and validation for the simultaneous determination of meloxicam and pridinol mesylate using RP-HPLC and its application in drug formulations.

A simple and reliable reversed-phase high-perfomance liquid chromatographic method has been developed and validated for the simultaneous determination of meloxicam and pridinol mesylate in their synthetic mixtures and combined tablet formulations. Both drugs were separated on a 250 mm x 4.6mm C18 column packed with 5 microm particles. The mobile phase, optimized through an experimental design, was a 51:9:40 (v/v/v) mixture of methanol, isopropanol and 50mM potassium phosphate buffer (pH 5.9), pumped at a flow rate of 1.0 ml min(-1). UV detection was performed at 225 nm. The method was validated in the sample concentration ranges of 33.7-61.8 mg l(-1) for meloxicam and 8.8-16.8 mg l(-1) for pridinol mesylate, where it demonstrated good linearity with r=0.9989 and 0.9987 (n=15), respectively. The assay was shown to be repeatable at concentration levels of 70%, 100% and 130%, with relative standard deviation values of 1.09% and 0.82% for meloxicam and pridinol, respectively. For independent 100% level samples, the intra-day precision was 0.4% and 1.0% while the intermediate precision was 0.7% and 1.0% for the drugs. The method demonstrated to be robust, resisting to small deliberate changes in pH, flow rate and composition (organic:aqueous ratio) of the mobile phase. The LOD values were 0.22 and 0.20 mg l(-1), while the LOQ were 1.7 and 1.1 mg l(-1), for meloxicam and pridinol, respectively. The applicability of the method was demonstrated by determining the drug content of two commercial pharmaceutical formulations, where it exhibited good performance.

A sensitive spectrophotometric method for the determination of desloratadine in tablets.

A simple, rapid, and sensitive visible spectrophotometric method was developed, for the first time, for analysis of desloratadine (DE) in tablets. The method is based on the deep-blue colored TCNQ*- radical anion formed by interaction of the drug (n-donor) with 7,7,8,8-tetracyanoquinodimethane (TCNQ, pi-acceptor) in acetonitrile at ambient temperature. Optimum conditions for the reaction were investigated, absorbances were read at 843 nm, and the linearity range for concentrations of DE was found to be 1.5-13 microg/mL. The reaction product remains stable up to 8 h when kept at room temperature in the dark. The developed method was validated and successfully applied to the determination of DE in tablets. The tablets were also analyzed with a column liquid chromatography method reported in literature. The results from both methods were statistically compared by t- and F-tests. No significant difference was found for the means and standard deviations at 95% confidence level. Accuracy was examined through recovery studies. Being very simple and reliable, the method can be recommended for routine quality control analysis of DE in tablets.

New ipratropium formulation to decrease nebulization time.

A new anticholinergic aerosol containing 0.5mg ipratropium bromide dissolved in 1mL of solution has been produced with the purpose of decreasing nebulization time for patients compared to the traditional formulation which is twice as voluminal (0.5mg/2mL, Boehringer-Ingelheim, France). The aim of this study was to compare aerosol characteristics (inhaled mass, particle size distribution and nebulization time) of these two formulations of ipratropium bromide, nebulized alone and with terbutaline (5mg/2mL, Astra Zeneca, Sweden), to determine whether the new formulation was equivalent to the old one. Four different jet nebulizers were used: PariLC+, Atomisor NL9M, Sidestream and Mistyneb. Statistical analysis of the results showed that for all types of nebulizer, the inhaled mass of ipratropium bromide 0.5mg/1mL was significantly lower than the inhaled mass of ipratropium bromide 0.5mg/2mL, and that there was no statistical difference between the inhaled mass of ipratropium bromide 0.5mg/1mL+terbutaline 5mg/2mL and the inhaled mass of ipratropium bromide 0.5mg/2mL+terbutaline 5mg/2mL. The study also showed that the new formulation of ipratropium bromide (0.5mg/1mL) mixed with terbutaline allowed a 26% decrease in nebulization time compared to the old formulation (0.5mg/2mL) mixed with terbutaline without changing aerosol characteristics (inhaled mass and particle size distribution). This leads to the conclusion that a 2mL minimum volume is required for nebulization, and that nebulization of ipratropium bromide 0.5mg/1mL alone must be avoided.

Determination of thiencynonate by liquid chromatographic-mass spectrometry and its application to pharmacokinetics in rats.

A sensitive and specific high-performance liquid chromatography-tandem mass spectrometry method (LC/ESI/MS) was developed and validated for the identification and quantification of the novel lead compound of anticholinergic drug thiencynonate in rat plasma. The analytes were determined using positive electrospray ionization mass spectrometry in the selected reaction ion monitoring (SRM). The chromatography separation was on BetaBasic-18 column (150 mm x 2.1 mm i.d., 3 microm). The mobile phase was composed of methanol-water (70:30, v/v), containing 0.5 per thousand formic acid, which was pumped at a flow rate of 0.2 ml/min. Phencynonate was selected as the internal standard (IS). Simultaneous MS detection of thiencynonate and IS was performed at m/z 364.4 (thiencynonate), m/z 358 (phencynonate), and the SRM of the two compounds were both at 156. Thiencynonate eluted at approximately 2.8 min, phencynonate eluted at approximately 2.9 min and no endogenous materials interfered with their measurement. Linearity was obtained over the concentration range of 1-100 ng/ml in rat plasma. The lower limit of quantification (LLOQ) was reproducible at 1 ng/ml in rat plasma. The precision measured was obtained from 2.47 to 9.28% in rat plasma. Extraction recoveries were in the range of 67.63-76.76% in plasma. This method was successfully applied to the identification and quantification of thiencynonate in pharmacokinetic studies.

Chemically modified carbon paste electrode for the potentiometric determination of dicylomine hydrochloride in batch and in FIA conditions.

A carbon-paste electrode for dicyclomine hydrochloride (DcCl) was prepared and fully characterized in terms of composition, life span, usable pH range and temperature. The electrode was applied to the potentiometric determination of dicylominium ion in its pure state and in pharmaceutical preparations in batch and flow injection conditions and in biological fluids using standard additions method. The electrode is based on a mixture of two ion-exchangers, namely, dicyclominium-phosphomolybdate and dicylominium-tetraphenylborate, dissolved in dioctyl phthalate as pasting liquid. The modified electrode showed a near-Nernstian slope of 58 +/- 2 mV over the concentration range of 1.2 x 10(-5)-1.6 x 10(-2) M with an average recovery of 97-102% and a RSD of 0.090-1.00. The electrode exhibits good selectivity for DcCl with respect to a large number of inorganic cations, sugars, amino acids and organic substances of biological fluids. Potentiometric titrations of DcCl with several titrants have been monitored using this modified carbon paste electrode as an end-point indicator electrode.

A critical appraisal of the utility of the serum anticholinergic activity assay in research and clinical practice.

The serum anticholinergic activity (SAA) assay was originally designed to quantify the anticholinergic burden of drug exposure. The same assay has been used to measure the anticholinergic activity of standard drug solutions. There are limitations to the use of the assay in research and in applying these findings to clinical practice. Assays of standard drug solutions do not account for pharmacokinetic differences among drugs, which limits the interpretation of such measurements. In addition, emerging evidence has suggested that anticholinergic medications may not be the only cause of elevated SAA. Despite these limitations, elevated SAA has been consistently associated with cognitive impairment and delirium in a number of research settings. Such findings have prompted investigators to consider the potential application of the SAA assay in research and in clinical practice. Therefore, the objectives of this review are to summarize the current literature involving the SAA assay, describe the relative merits and shortfalls of the SAA assay as a research tool, and discuss the potential for use of the SAA assay as a clinical tool.