Hemp seed (Cannabis sativa L.) contain large amounts of nutrients, e.g. protein, dietary fiber, minerals, and unsaturated fatty acids, which make them a good fortifying component in food production. The aim of the present study was to determine the effect of hemp addition on the physicochemical properties, cooking quality, texture parameters and sensory properties of durum wheat pasta. The samples were fortified with 5–40% of commercially available hemp flour or 2.5–10% of hemp cake obtained from hemp seed oil pressing. Our study showed that the addition of hemp seed raw materials led to an increase in the protein, total dietary fiber (TDF), ash and fat content in the pasta samples. Due to its lower granulation and higher nutritional value, hemp flour was found to be a better raw material for the fortification of pasta than hemp cake. Pasta enriched with hemp flour at the level of 30–40% contains 19.53–28.87% d.m. of protein and 17.02–21.49% d.m. of TDF and according to the EU, a definition can be described as a high-protein and high-fiber products. All enriched pasta samples were also characterized by safe Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) content, and their sensory properties were accepted by consumers.

Background: Identification and quantitative determination of cytisine, especially in biological samples and pharmaceutical formulations, is still a difficult analytical task. Cytisine is an alkaloid with a small and very polar molecule. For this reason, it is very weakly retained on reversed phase (RP) stationary phases, such as commonly used alkyl-bonded phases. The very weak retention of cytisine causes it to be eluted together with the components of biological matrices. Objective: Comparison and evaluation of various chromatographic systems for analysis of cytisine in different matrices—serum, saliva and pharmaceutical formulation—by high performance liquid chromatography (HPLC) with diode array (DAD), fluorescence (FLD) and mass spectrometry (MS) detection. Methods: The analyses were performed using HPLC in reversed phase (RP), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) modes. Different sample pre-treatment methods were tested: Protein precipitation (with acetone, methanol (MeOH) or acetonitrile (ACN), and solid phase extraction (SPE) using cartridges with octadecyl (C18), hydrophilic-lipophilic balanced copolymer (HLB) or strong cation exchange sorbents (Strata X-C). Conclusion: Significant differences were observed in retention parameters with a change of the used chromatographic system. The various properties of stationary phases resulted in differences in analyte retention, peaks’ shape and systems’ efficiency. The weakest retention was observed using RP systems; however, the use of the Polar RP phase can be an alternative for application in green chromatography. In the strongest retention was observed using a strong cation exchange (SCX) phase. The most optimal systems were chosen for the analysis of cytisine in the pharmaceutical preparation, serum and saliva after sample pre-treatment with the new SPE procedure. Due to the sensitivity, the use of HPLC-DAD or HPLC-FLD is the most optimal for drug analysis in pharmaceutical preparations, whereas HPLC-MS is suitable for analysis of cytisine in biological samples.

The routine techniques currently applied for the determination of nicotine and its major metabolites, cotinine, and trans-3′-hydroxycotinine, in biological fluids, include spectrophotometric, immunoassays, and chromatographic techniques. The aim of this study was to develop, and compare two new chromatographic methods high-performance liquid chromatography coupled to triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS), and RP-HPLC enriched with chaotropic additives, which would allow reliable confirmation of tobacco smoke exposure in toxicological and epidemiological studies. The concentrations of analytes were determined in human plasma as the sample matrix. The methods were compared in terms of the linearity, accuracy, repeatability, detection and quantification limits (LOD and LOQ), and recovery. The obtained validation parameters met the ICH requirements for both proposed procedures. However, the limits of detection (LOD) were much better for HPLC-QQQ-MS/MS (0.07 ng mL−1 for trans-3′-hydroxcotinine; 0.02 ng mL−1 for cotinine; 0.04 ng mL−1 for nicotine) in comparison to the RP-HPLC-DAD enriched with chaotropic additives (1.47 ng mL−1 for trans-3′-hydroxcotinine; 1.59 ng mL−1 for cotinine; 1.50 ng mL−1 for nicotine). The extraction efficiency (%) was concentration-dependent and ranged between 96.66% and 99.39% for RP-HPLC-DAD and 76.8% to 96.4% for HPLC-QQQ-MS/MS. The usefulness of the elaborated analytical methods was checked on the example of the analysis of a blood sample taken from a tobacco smoker. The nicotine, cotinine, and trans-3′-hydroxycotinine contents in the smoker’s plasma quantified by the RP-HPLC-DAD method differed from the values measured by the HPLC-QQQ-MS/MS. However, the relative errors of measurements were smaller than 10% (6.80%, 6.72%, 2.04% respectively).

Retention, separation selectivity and system efficiency of selected basic psychotropic drugs (clozapine, aripiprazole, vortioxetine and zolpidem) and drug metabolites (desmethylclozapine, clozapine N-oxide and dehydroaripiprazole) on Hydro RP, Phenyl-Hexyl and Polar RP columns were studied. Mobile phases containing methanol or acetonitrile as organic modifiers, acetate buffer at pH 3.5 and addition of diethylamine (DEA) as a silanol blocker were applied. Significant differences in the retention, peak shapes and systems' efficiency of the investigated compounds were obtained depending on the tested chemically bonded stationary phases with various ligands. Based on the obtained results the Phenyl-Hexyl column was selected for analysis of the drugs and their metabolites in human serum and saliva samples.Solid phase extraction (SPE) was applied for sample pretreatment. The best SPE-HPLC-DAD procedure was used for simultaneous analysis of clozapine, aripiprazole and their metabolites in body fluids. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was applied for confirmation of the presence of the investigated compounds in biological samples. The lower limit of quantification (LLOQ) of clozapine obtained using the proposed method was 10 ng/mL. The validated method for determining the presence of clozapine and its main metabolite was successfully applied in therapeutic drug monitoring.

Selection of livestock has not only led to changes in the level of their performance but also modified their behavior. As a result, within a single species, we have to deal with different behaviors of different breeds. In our study, we assumed that the different behaviors within a species are due to differences in the morphology and physiology of behavior-related systems. Two breeds of hens were used as a model: the highly reactive, fearful and high-performance Leghorn breed and proactive, unselected Green-legged Partridge breed. The higher reactivity and fearfulness of Leghorn hens in comparison to the Green-legged Partridge breed may be related to the greater number of neurons in the paraventricular nucleus and anterior hypothalamus and the higher content of zinc and iron in the brain, as these elements are involved in neuronal conduction and myelination processes. The reactive behaviours of Green-legged Partridge hens may be associated with the lower number of neurons in the paraventricular nucleus and the anterior hypothalamus and the higher concentration of dopamine and copper ions in the brain. The analyses confirmed the hypothesis of the existence of interbreed differences in the morphology and physiology of behaviour-related systems, which most probably emerged through unintentional and correlated selection towards high production performance. Consequently, attention should be drawn that the selection of a given genotype (breed) towards a specific environment could lead to creation of highly specialised lines that may not achieve homeostasis in every maintenance system.