BACKGROUND: Strigolactones represent the most recently described group of plant hormones involved in many aspects of plant growth regulation. Simultaneously, root exuded strigolactones mediate rhizosphere signaling towards beneficial arbuscular mycorrhizal fungi, but also attract parasitic plants. The seed germination of parasitic plants induced by host strigolactones leads to serious agricultural problems worldwide. More insight in these signaling molecules is hampered by their extremely low concentrations in complex soil and plant tissue matrices, as well as their instability. So far, the combination of tailored isolation-that would replace current unspecific, time-consuming and labour-intensive processing of large samples-and a highly sensitive method for the simultaneous profiling of a broad spectrum of strigolactones has not been reported. RESULTS: Depending on the sample matrix, two different strategies for the rapid extraction of the seven structurally similar strigolactones and highly efficient single-step pre-concentration on polymeric RP SPE sorbent were developed and validated. Compared to conventional methods, controlled temperature during the extraction and the addition of an organic modifier (acetonitrile, acetone) to the extraction solvent helped to tailor strigolactone isolation from low initial amounts of root tissue (150 mg fresh weight, FW) and root exudate (20 ml), which improved both strigolactone stability and sample purity. We have designed an efficient UHPLC separation with sensitive MS/MS detection for simultaneous analysis of seven natural strigolactones including their biosynthetic precursors-carlactone and carlactonoic acid. In combination with the optimized UHPLC-MS/MS method, attomolar detection limits were achieved. The new method allowed successful profiling of seven strigolactones in small exudate and root tissue samples of four different agriculturally important plant species-sorghum, rice, pea and tomato. CONCLUSION: The established method provides efficient strigolactone extraction with aqueous mixtures of less nucleophilic organic solvents from small root tissue and root exudate samples, in combination with rapid single-step pre-concentration. This method improves strigolactone stability and eliminates the co-extraction and signal of matrix-associated contaminants during the final UHPLC-MS/MS analysis with an electrospray interface, which dramatically increases the overall sensitivity of the analysis. We show that the method can be applied to a variety of plant species.

• Klíčová slova
• Phosphate starvation, Phytohormones, Quantitative analysis, Solid phase extraction (SPE), Strigolactones, UHPLC–MS/MS,
• Publikační typ
• časopisecké články MeSH

Stress-induced changes in phytohormone metabolite profiles have rapid effects on plant metabolic activity and growth. The jasmonates (JAs) are a group of fatty acid-derived stress response regulators with roles in numerous developmental processes. To elucidate their dual regulatory effects, which overlap with those of other important defence-signalling plant hormones such as salicylic acid (SA), abscisic acid (ABA) and indole-3-acetic acid (IAA), we have developed a highly efficient single-step clean-up procedure for their enrichment from complex plant matrices that enables their sensitive quantitative analysis using hyphenated mass spectrometry technique. The rapid extraction of minute quantities of plant material (less than 20mg fresh weight, FW) into cold 10% methanol followed by one-step reversed-phase polymer-based solid phase extraction significantly reduced matrix effects and increased the recovery of labile JA analytes. This extraction and purification protocol was paired with a highly sensitive and validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method and used to simultaneously profile sixteen stress-induced phytohormones in minute plant material samples, including endogenous JA, several of its biosynthetic precursors and derivatives, as well as SA, ABA and IAA.

• Klíčová slova
• Abscisic acid, Arabidopsis thaliana, Indole-3-acetic acid, Jasmonates, Salicylic acid, Solid-phase extraction (SPE), Stress-induced phytohormones, Tandem mass spectrometry (MS/MS), Ultra-high performance liquid chromatography (UHPLC),
• MeSH
• chromatografie kapalinová MeSH
• cyklopentany analýza   chemie   MeSH
• extrakce na pevné fázi MeSH
• fyziologický stres MeSH
• kyselina abscisová analýza   chemie   MeSH
• kyselina salicylová analýza   chemie   MeSH
• kyseliny indoloctové analýza   MeSH
• listy rostlin chemie   MeSH
• mastné kyseliny analýza   chemie   MeSH
• molekulární struktura MeSH
• oxylipiny analýza   chemie   MeSH
• regulátory růstu rostlin chemie   izolace a purifikace   fyziologie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cyklopentany MeSH
• indoleacetic acid MeSH Prohlížeč
• jasmonic acid MeSH Prohlížeč
• kyselina abscisová MeSH
• kyselina salicylová MeSH
• kyseliny indoloctové MeSH
• mastné kyseliny MeSH
• oxylipiny MeSH
• regulátory růstu rostlin MeSH

BACKGROUND: Statins are the microsomal 3-hydroxy-3methylglutaryl-coenzyme A reductase inhibitors used for the treatment of hypercholesterolemia. Some recent studies revealed also the extra-lipid effects and anticancer activities. Due to the wide incidence of cancer diseases, the number of studies dealing with anticancer statin activities has grown in recent years. Development of one universal multistatin method will be a very convenient way of providing practical and economical multiple statin analysis. Results/methodology: Fast and sensitive methods for determination of seven clinically relevant statins, their interconversion products and metabolites (17 analytes in total) in biological samples using microextraction by packed sorbent for sample preparation and UHPLC-MS/MS for subsequent analysis were developed and validated. Three MS platforms with different ion sources, transfer optics, collision cell technologies and scan speed parameters were compared. CONCLUSION: Significant differences among the methods were observed in terms of selectivity and sensitivity. Microextraction by packed sorbent was successful in the extraction of all 17 analytes from biological matrix.

• Klíčová slova
• MEPS, SPE, UHPLC–MS/MS, bioanalysis, dwell time, interconversion, multianalyte, protein precipitation, statins, triple quadrupole systems,
• MeSH
• biochemická analýza krve metody   MeSH
• chemická precipitace MeSH
• lidé MeSH
• limita detekce MeSH
• mikroextrakce na pevné fázi metody   MeSH
• reprodukovatelnost výsledků MeSH
• statiny krev   chemie   izolace a purifikace   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• statiny MeSH

The new ultra-high performance liquid chromatography method with tandem mass spectrometry detection (UHPLC-MS/MS) has been optimized to allow fast, selective, and high-throughput analysis of two Candida albicans quorum sensing molecules (QSM), farnesol and tyrosol. The problem of the presence of the interference in the samples and system was successfully solved by careful optimization of chromatographic conditions. Charged hybrid stationary phase modified with pentafluorophenyl group and optimized gradient elution provided adequate separation selectivity and peak shapes. The impurity was identified as dibutyl phthalate and had the same m/z ions as farnesol leading to an important interference on selected reaction monitoring channel. Two different types of biological matrices originating from vaginal fluid, supernatant and sediment, were analysed. Micro-solid phase extraction in pipette tips was optimized for the selective isolation of QSM from the supernatant. The insufficient retention of farnesol on the extraction sorbent was improved when 1% of organic solvent was added prior to extraction, while the retention of tyrosol was only possible when using combined C8 and polymer sorbent type. Strong retention of farnesol had to be solved by increasing elution solvent strength and volume up to 600 μL. However, this approach did not allow the pretreatment of sediment samples due to the sorbent clogging. Therefore, our previously developed protein precipitation method was modified and validated to analyse the sediments. New developed UHPLC-MS/MS method provided suitable accuracy and precision for the determination of QSM in vaginal fluid while using only 50 μL sample volume and two different sample preparation methods.

• Klíčová slova
• Candida albicans, Farnesol, Microextraction, Quorum sensing, Tyrosol, UHPLC-MS/MS,
• MeSH
• Candida albicans izolace a purifikace   MeSH
• dospělí MeSH
• farnesol analýza   MeSH
• fenethylalkohol analogy a deriváty   analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• limita detekce MeSH
• mladý dospělý MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vagina chemie   mikrobiologie   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Názvy látek
• 4-hydroxyphenylethanol MeSH Prohlížeč
• farnesol MeSH
• fenethylalkohol MeSH

While the prudent and reasonable use of veterinary antimicrobial agents in food-producing animals is necessary, researchers over the decades have shown that these antimicrobial agents can spread into the environment through livestock manure and wastewater. The analysis of the occurrence of antimicrobial compounds in soil samples is of a great importance to determine potential impacts on human and animal health and the environment. In this study, an affordable, rugged and simple analytical method has been developed for the determination of twenty-nine antimicrobial compounds from five different classes (tetracyclines, fluoro(quinolones), macrolides, sulfonamides and diaminopirimidines). Liquid-liquid extraction (LLE) with extract filtration combined with ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was the best strategy for the simultaneous determination of all analytes. The developed method was validated according to the Commission Implementing Regulation (EU) 2021/808. The limit of detections (LODs) ranged from 0.5 to 2.0 µg/kg, while the limit of quantitation (LOQ) was established at 1.0 to 20.0 µg/kg. The developed method was successfully applied for the determination of antimicrobial residues in one hundred and eighteen soil samples obtained from four European countries (Austria, Czech Republic, Estonia and Portugal). Doxycycline in the concentration levels of 9.07 µg/kg-20.6 µg/kg was detected in eight of the analysed samples. Samples were collected from areas where natural fertilizers (swine or cow manure) were applied. Our method can be efficiently used to monitor anti-microbial compounds in soil samples.

• Klíčová slova
• UHPLC-MS/MS, antimicrobial agents, environment, soil,
• MeSH
• antibakteriální látky analýza   MeSH
• antiinfekční látky * MeSH
• extrakce na pevné fázi MeSH
• hnůj analýza   MeSH
• lidé MeSH
• prasata MeSH
• půda MeSH
• skot MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• antiinfekční látky * MeSH
• hnůj MeSH
• půda MeSH

An automated flow analysis-solid phase extraction (FA-SPE) system and methodology of ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) analysis were developed for the determination of selected antiviral drugs (acyclovir, amantadine, rimantadine, and oseltamivir) in water samples. The proposed FA-SPE approach enables the integration of various extraction stages and elimination of the sample evaporation step and offers individual customisation of SPE parameters, inter alia sample, and eluate flow rate and volume. Using the developed FA-SPE procedure, e.g. a 100-fold preconcentration of the target analytes in 1 h was achieved. A method for chromatographic analysis was also developed to determine the selected antiviral drugs in combination with the use of the FA-SPE system. The developed FA-SPE UHPLC-MS/MS method was validated including the determination of linearity of analytical graphs, limits of detection (5.5-99.9 pg mL-1) and quantification (18.3-329.8 pg mL-1), intra-day (1.8-8.3%) and inter-day (3.0-9.2%) precision, recovery (95.6-105.3%), and matrix effects (- 12.9 to 13.2%). The proposed method was successfully applied to analyse tap, drinking, and river water samples, revealing the presence of amantadine at a concentration of 40.1 pg mL-1 in one sample. The environmental impact of the developed FA-SPE sample preparation procedure was also assessed using the AGREEprep metric tool and compared with five other literature methods, achieving the most sustainable outcome.

• Klíčová slova
• Antiviral drugs, Emerging contaminants, Flow analysis, Solid phase extraction, UHPLC-MS/MS, Water analysis,
• MeSH
• antivirové látky * analýza   MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• extrakce na pevné fázi * MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• voda chemie   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antivirové látky * MeSH
• chemické látky znečišťující vodu * MeSH
• voda MeSH

Differences among lipidomic profiles of healthy volunteers, obese people and three groups of cardiovascular disease (CVD) patients are investigated with the goal to differentiate individual groups based on the multivariate data analysis (MDA) of lipidomic data from plasma, erythrocytes and lipoprotein fractions of more than 50 subjects. Hydrophilic interaction liquid chromatography on ultrahigh-performance liquid chromatography (HILIC-UHPLC) column coupled with electrospray ionization mass spectrometry (ESI-MS) is used for the quantitation of four classes of polar lipids (phosphatidylethanolamines, phosphatidylcholines, sphingomyelins and lysophosphatidylcholines), normal-phase UHPLC-atmospheric pressure chemical ionization MS (NP-UHPLC/APCI-MS) is applied for the quantitation of five classes of nonpolar lipids (cholesteryl esters, triacylglycerols, sterols, 1,3-diacylglycerols and 1,2-diacylglycerols) and the potential of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is tested for the fast screening of all lipids without a chromatographic separation. Obtained results are processed by unsupervised (principal component analysis) and supervised (orthogonal partial least squares) MDA approaches to highlight the largest differences among individual groups and to identify lipid molecules with the highest impact on the group differentiation.

• Klíčová slova
• Cardiovascular diseases, Lipidomics, Lipids, Lipoprotein fractions, Multivariate data analysis, UHPLC/MS,
• MeSH
• dospělí MeSH
• kardiovaskulární nemoci krev   MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipidy krev   chemie   klasifikace   MeSH
• lipoproteiny krev   chemie   klasifikace   MeSH
• metoda nejmenších čtverců MeSH
• multivariační analýza MeSH
• obezita MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• výpočetní biologie MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• lipidy MeSH
• lipoproteiny MeSH

Certain agricultural plastics, i.e., mulching films, are generally considered as potent sources of micro- and nanoplastics (MNPs), due to their direct application on soil and waste mishandling. During the synthesis and fabrication of such agricultural plastics, it is necessary to use chemicals, the so-called plastic additives (PAs), improving the physicochemical properties of the final polymeric product. However, since PAs are loosely bound on the polymer matrix, they can potentially leach into the soil environment with unidentified effects. Clearly, to monitor the fate of PAs in the terrestrial ecosystem, it is necessary to develop accurate, sensitive and robust analytical methods. To this end, a comprehensive analytical strategy was developed for monitoring 16 PAs with diverse physicochemical properties (partition coefficient; -3 < logP<19) in soil samples using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). For this purpose, two different extraction procedures were developed, namely, a single step ultrasound-assisted extraction (UAE) using ethyl acetate or an aqueous solution of methanol and a binary extraction, combining Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) and UAE principles with n-hexane as the extractant. Interestingly, within the sample preparation investigation, we identified in-lab contamination sources of PAs, e.g., centrifuge tubes or microfilters. Such consumables are made of plastic contaminating the procedural blanks and omitting their use was necessary to acquire satisfactory analytical performance. In detail, method validation was performed for 16 compounds achieving recoveries mainly in the range 70-120 %, repeatability (expressed as relative standard deviation, RSD %) < 20 % and limits of quantification (LOQs) ranging between 0.2 and 20 ng/g dry weight (dw). Importantly, the presented strategies are added to the very limited available for PA determination in soil, a topical issue with a significant and rather understudied impact on agriculture.

• Klíčová slova
• Agricultural plastics, Plastic additives, Plastic pollution, Quality assurance, Soil, UHPLC-MS/MS,
• MeSH
• látky znečišťující půdu * analýza   MeSH
• monitorování životního prostředí metody   MeSH
• plastické hmoty * analýza   MeSH
• půda * chemie   MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zemědělství * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• látky znečišťující půdu * MeSH
• plastické hmoty * MeSH
• půda * MeSH

Management of the therapy of life-threatening bacterial infection is extremely based on an optimal antibiotic treatment. Achieving the correct vancomycin dosage in blood and target tissues can be complicated in special situations, e.g., where large fluid sequestration and/or acute renal failure occur. A UHPLC-MS/MS method operating in electrospray (ESI) positive ion mode was applied for the determination of vancomycin in serum, urine and peritoneal/pleural effusion. Sample pretreatment was composed of dilution and simple protein precipitation where only a small volume (50μL) of serum, urine or peritoneal/pleural effusion was required. The separation of vancomycin was performed on a Meteoric Core C18 BIO column (100×4.6mm, 2.7μm) by gradient elution with 0.1% formic acid in water and acetonitrile. The total time of analysis was 4.5min. The method was found to be linear in the range of 2-60μM (or 0.5-10μM) for serum, 0.27-10μM (or 2-60μM) for peritoneal/pleural effusion and 25-300μM for urine, which was adequate for the determination of vancomycin in patient samples. The intra- and inter-day precision was below 8% RSD, and accuracy was from 89 to 104%. The UHPLC/MS-MS method offers a fast and reliable approach to determine vancomycin concentrations in three different human body fluid samples (serum, urine and peritoneal/pleural effusion) with a simple sample pretreatment that was the same for all selected specimens. This method should be applicable to large sample series in clinical (pharmacokinetic/pharmacodynamic) studies.

• Klíčová slova
• Peritoneal/pleural effusion, Serum, UHPLC–MS/MS, Urine, Vancomycin,
• MeSH
• antibakteriální látky MeSH
• ascitická tekutina * MeSH
• lidé MeSH
• pleurální výpotek * MeSH
• tandemová hmotnostní spektrometrie MeSH
• vankomycin MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• vankomycin MeSH

A complete analytical procedure combining optimized tea infusion preparation and validated UHPLC-MS/MS method was developed for routine quantification of eight naturally occurring catechin derivatives in various tea samples. The preparation of tea infusions was optimized in terms of temperature, time and water-to-tea ratio in green, white and black teas. The catechins were analyzed using ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry in a run of only 4 min including equilibration of the system. The UHPLC-MS/MS method was fully validated in terms of inter/intra-day precision, accuracy, linearity (r(2)>0.9991), range (50-5000 ng/ml), LOD (1.5-7.5 ng/ml) and LOQ (5-25 ng/ml). Validation of the method included also the determination of the matrix effects that were evaluated in both flavored and unflavored green, white and black teas. Dilution of the resulting tea infusions appeared to be crucial for the matrix effects and also for subsequent catechin quantification in real tea samples in order to fit into the linear range of the UHPLC-MS/MS method. This complete procedure for catechin quantification was finally applied to real sample analysis represented by 70 commercial tea samples.

• Klíčová slova
• Catechins, Matrix effects, Method validation, Tea, UHPLC–MS/MS,
• MeSH
• analýza potravin metody   MeSH
• čaj chemie   MeSH
• katechin chemie   MeSH
• limita detekce MeSH
• reprodukovatelnost výsledků MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• teplota MeSH
• voda chemie   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• validační studie MeSH
• Názvy látek
• čaj MeSH
• katechin MeSH
• voda MeSH