LC and LC-MS/MS Studies for the Separation and Identification of Impurities and Degradation Products of Betaxolol.

Betaxolol (Bx) is a selective ß1 receptor blocker used in the treatment of hypertension and glaucoma. The aim of the present work was to demonstrate an approach involving use of liquid chromatography (LC) and liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS) for the simultaneous separation, identification and characterization of impurities and of degradation products of betaxolol without their isolation from the reaction mixtures. At optimum condition, and according to ICH guidelines, the limit of detection (LOD) and limit of quantification (LOQ) for Bx are found to be 5.46 and 16.54 µg mL-1, respectively. However, the LOD and LOQ for the major degradation product P6 were 2.15 and 6.53 µg mL-1. Betaxolol was subjected to hydrolytic (acidic and basic) and oxidative, stress conditions according to International Conference on Harmonization (ICH) guideline Q1A (R2), and as results, the drug was found to be labile in acidic, basic and oxidative stress conditions. Based on LC-ESI/MS analysis, the found results revealed that Bx decomposes in acidic, basic and oxidizing environments. All degradation products were identified with the help of their fragmentation pattern and the masses obtained upon the MS analysis.

Assessment of trace levels of aflatoxins AFB1 and AFB2 in non-dairy beverages by molecularly imprinted polymer based micro solid-phase extraction and liquid chromatography-tandem mass spectrometry.

A selective molecularly imprinted polymer (MIP) adsorbent was synthesised and used in a batch micro-solid phase extraction format for isolating aflatoxins (AFB1, and AFB2) from non-dairy beverages before liquid chromatography-tandem mass spectrometry determination. MIP synthesis (precipitation polymerization in 3 : 1 acetonitrile/toluene as a porogen) was performed with 5,7-dimethoxycoumarin (DMC), methacrylic acid (MAA) and divinylbenzene-80 (DVB) as a dummy template, functional monomer and cross-linker, respectively (1 : 4 : 20 molar ratio). 2,2'-Azobisisobutyronitrile (AIBN) was used as a polymerization initiator. The adsorbent MIP (50 mg) was enclosed in a cone-shaped polypropylene membrane (porous membrane protected molecularly imprinted micro-solid phase extraction), and parameters such as sample pH, mechanical (orbital-horizontal) shaking, the extraction time (loading stage), the composition of the eluting solution, and the desorption time were optimised. The highest extraction yields were obtained by using 5 mL of non-dairy beverages (pH adjusted at 6.0), and mechanical shaking (150 rpm) for 15 min. Elution was performed with 5 mL of an acetonitrile/formic acid (97.5 : 2.5) mixture under ultrasound (325 W, 35 kHz) for 15 min. After eluate evaporation to dryness and re-dissolution in 150 µL of the mobile phase, the pre-concentration factor of the method was 33.3, which yields limits of detection within the 0.085-0.207 µg L-1 range. In addition, the current proposal was shown to be an accurate and precise method through relative standard deviation of intraday and inter-day assays below 18% and analytical recoveries in the range of 91-104%. However, the method was found to suffer from matrix effects.

Analytical strategy for the detection of ecdysterone and its metabolites in vivo in uPA(+/+)-SCID mice with humanized liver, human urine samples, and estimation of prevalence of its use in anti-doping samples.

Ecdysteroids are of interest as potential sport performance enhancers, due to their anabolic effects. The current study aimed to analyze levels of the most abundant ecdysteroid, ecdysterone (20-hydroxyecdysone, 20-OHE) in easily available dietary supplements, and, outline an analytical strategy for its detection, and that, of its metabolites, (1) following administration of pure 20-OHE to uPA(+/+)-SCID mice with humanized liver, (2) in a human volunteer after ingestion of two supplements, one with a relatively low, and the other a high, concentration of 20-OHE, and, (3) to estimate the prevalence of use of 20-OHE in elite athletes (n = 1000). Of the 16 supplements tested, only five showed detectable levels of 20-OHE, with concentrations ranging from undetectable up to 2.3 mg per capsule. Urine of uPA(+/+)-SCID urine showed the presence of 20-OHE and its metabolite, 14 deoxy ecdysterone, within 24 hours (hr) of ingestion. In humans, both the parent and the metabolite were detectable within 2 to 5 hr of ingestion, with the metabolite being detectable for longer than the parent. After ingestion of a low dose supplement, the parent and metabolite were detectable for 70 and 48 hr, while following the higher dose it was 96 and 48 hr, respectively. Analysis of urines from athletes (n = 1000) confirmed four positives for 20-OHE, suggesting a prevalence of use of 0.4%. Prevalence of its use by elite athletes was relatively low, however, this needs to be confirmed in other populations, and with other related ecdysteroids.

Aflatoxins screening in non-dairy beverages by Mn-doped ZnS quantum dots - Molecularly imprinted polymer fluorescent probe.

Synthesized molecularly imprinted polymer (MIP) materials have been anchored on the surface of PEG-Mn-doped ZnS quantum dots (QDs) to develop a fluorescence probe for aflatoxins (AFs) recognition/determination in non-dairy beverages. MIP synthesis used 5,7-dimethoxycoumarin (DMC) as a dummy template molecule, and methacrylic acid (MAA) as a functional monomer. Under optimized conditions (1.25 mL of 5 mg L-1 MIP-PEG-ZnS QDs solution, pH 5.0, and 12 min delay time before scanning), the prepared MIP-QDs composite was found to offer high affinity and selectivity for AFs (AFB1, AFB2, AFG1 and AFG2). A fast fluorimetric screening method (total AFs assessment) was therefore feasible. The limits of detection (LOD) and quantification (LOQ) were 0.016 and 0.053 mg L-1, respectively. Analytical recoveries (inter- and intra-day assays) were from 99 ±â€¯4% to 107 ±â€¯5%, with RSD (intra-day assay) lower than 13, and RSD (inter-day) lower than 7%. The optimized method was applied for total AFs assessment in several non-dairy beverage samples.

Organochlorine pesticides and polychlorinated biphenyls in human adipose tissue from northern Tunisia: Current extent of contamination and contributions of socio-demographic characteristics and dietary habits.

The aims of the present study were to investigate the current exposure levels of persistent organochlorine compounds (OCs) in adipose tissues intraoperatively collected from 40 patients over 20 years undergoing non-cancer-related surgery residing in Northern region of Tunisia (Bizerte), which constitutes an exemplary case, and examined association between levels of contamination and both socio-demographic characteristics and dietary habits. Concentration of hexachlorobenzene (HCB), hexachlorocyclohexane isomers (α-HCH, ß-HCH, γ-HCH and δ-HCH), dichlorodiphenyltrichloroethane isomers (p,p'-DDT and o,p'-DDT) and metabolites (p,p'-DDE, o,p'-DDE, p,p'-DDD and o,p'-DDD) and 12 polychlorinated biphenyls (PCBs) congeners were measured using capillary gas chromatography with electron capture detector. Overall, residue levels of OCs followed the decreasing order of DDTs > PCBs > HCB > HCHs. DDTs levels ranged from 74.49 to 1834.76ngg-1 lipid and contributing to more than 90% to the sum of organochlorine pesticides (OCPs). p,p'-DDE was the most abundant in all samples and the p,p'-DDT/p,p'-DDE ratio (range between 1.85% and 58.45%) suggesting recent and ongoing exposure to banned commercial DDT products. PCB concentrations varied from 29.27 to 322.58ngg-1 lipid and PCB-180, PCB-153 and PCB-138 were the dominant congeners accounting for 70% of total PCBs. We did not find significant correlations between OC exposure levels and sex, parity, habitat areas and smoking habits. In females, the adipose tissue concentrations of DDTs, HCB and PCB-118 were positively correlated with age. There was statistically significant relationship between body mass index (BMI) changes and the adipose tissue levels of HCB and HCHs. No association was found between OCPs levels and dietary factors. However, our study suggests that fish consumption may be an important contributor of PCBs adipose tissue content of PCBs in Tunisian people. The presented work is highly significant, being the first study pointing out the chronic exposure to OCs in Bizerte.

Study of a conformational equilibrium of lisinopril by HPLC, NMR, and DFT.

The isomerization of lisinopril has been investigated using chromatographic, NMR spectroscopic, and theoretical calculations. The NMR data, particularly the NOEDIFF experiments, show that the major species that was eluted first is the trans form. The proportion was 77% and 23% for the trans and cis, respectively. The thermodynamic parameters (ΔH, ΔS, and ΔG) were determined by varying the temperature in the NMR experiments. The interpretations of the experimental data were further supported by DFT/B3LYP calculations.