Chromatographic Trace Research Articles

Storing liquid chromatographic separations on surface energy traps: decoupling the LC and the mass spectrometer.

We report a micro-fractionation device for high performance liquid chromatography-mass spectrometry to archive chromatographic separations on an array of optimized surface energy traps (SETs). The method has the potential to significantly alter nanoflow LC-MS workflow, decoupling separation and analysis. The wetting characteristics of the SETs cause the HPLC eluent stream to spontaneously split into droplet microfractions. The droplet mirofractions are then dried down to enable facile storage and transport of the archived separation. Discontinuously dewetting array parameters were explored to maximize array volume and resolution using a combination of SET design, shape, size, and spacing. Mass spectrometry analysis is performed utilizing a liquid micro-junction surface sampling probe to extract dried analytes from the surface of the SETs followed by electrospray ionisation. A reverse phase separation of pharmaceutical compounds is "recorded" using the micro-fractionation device followed by "reading" the chromatographic trace with a mass spectrometer 24 hours after the separation was performed/archived, demonstrating a true decoupling of LC, and MS. Additionally, we demonstrate the ability to collect microfractions with sub-one-second integration time, approaching the scan time of a mass spectrometer or UV-Vis detector. With further improvements to the device, sub-1-second micro-fractionation may enable seamless reconstruction of archived chromatograms indistinguishable from online LC-MS data, while also providing the benefits of easy storage and transport of archived separations.

A study on the impact of harvesting operations on the mineral oil contamination of olive oils

During the 2020–21 olive oil campaign, the contribution of harvesting operations to mineral oil saturated (MOSH) and aromatic hydrocarbon (MOAH) contamination was studied. Oils extracted from hand-picked olives (15 different olive groves) generally had background MOSH (<2.7 mg/kg), and no quantifiable MOAH. In 40% of the cases, an important contamination increase was observed after harvesting operations. Except for one sample (325.8 and 111.0 mg/kg of MOSH and MOAH, respectively), other samples reached 4.3–33.7 mg/kg of MOSH and 1.1–11.3 mg/kg of MOAH. Accidental leaks of lubricants and/or contact with lubricated mechanical parts, were identified as important sources of contamination. Chromatographic traces obtained by on-line high-performance liquid chromatography (HPLC)-gas chromatography (GC)-flame ionization detection (FID) allowed for source identification. A comprehensive two-dimensional gas chromatographic platform (GC × GC) with parallel FID/MS detection was implemented for confirmation and to attempt the characterization of the contaminations. Good harvesting practices are suggested to minimize contamination risks.

Non-psychoactive cannabinoids identification by linear retention index approach applied to a hand-portable capillary liquid chromatography platform.

The aim of the present research was the application of the linear retention index (LRI) system for the identification of non-psychoactive cannabinoids using a portable LC instrument. The miniaturization, viz. the use of very low quantities of mobile phase, enabled the development of a compact mobile system to be used for in situ analysis, also according to a green and cost-saving approach. In particular, new capillary LC (cap-LC) methods coupled with UV detection were developed for the analysis of extracts of Cannabis sativa L. Two setups were explored to achieve the efficient separation of twenty-four cannabinoids: a single column setup which exploited a sub-2µm packing to increase the chromatographic resolution, and a dual-column setup based on the serial connection of two different stationary phases, each coupled to an UV detector. The latter allowed the determination of two LRI values for each analyte, thus increasing the identification power. Moreover, since two different wavelengths were used on the LED-based UV detectors, the ratio of the absorbances measured on each chromatographic trace represented a third identification criterion, thus fulfilling the recommendations of the Scientific Working Group for The Analysis of Seized Drugs (SWDRUG) about the categories of analytical techniques to be used and the minimum number of parameters required for the unambiguous identification of drugs. The obtained results could be used for the development of a novel analytical method for fast and automatic in situ forensic investigations and hemp breeding programs, also minimizing the consumption of both sample and solvent.

Identification and Relative Quantification of hFSH Glycoforms in Women's Sera via MS-PRM-Based Approach.

Follicle-stimulating hormone (FSH) is a glycohormone synthesized by adenohypophysis, and it stimulates ovulation in women and spermatogenesis in men by binding to its receptor (FSHR). FSHR is involved in several mechanisms to transduce intracellular signals in response to the FSH stimulus. Exogenous FSH is currently used in the clinic for ovarian hyperstimulation during in vitro fertilization in women, and for treatment of infertility caused by gonadotropin deficiency in men. The glycosylation of FSH strongly affects the binding affinity to its receptor, hence significantly influencing the biological activity of the hormone. Therefore, the accurate measurement and characterization of serum hFSH glycoforms will contribute to elucidating the complex mechanism of action by which different glycoforms elicit distinct biological activity. Nowadays ELISA is the official method with which to monitor serum hFSH, but the test is unable to distinguish between the different FSH glycovariants and is therefore unsuitable to study the biological activity of this hormone. This study presents a preliminary alternative strategy for identifying and quantifying serum hFSH glycoforms based on immunopurification assay and mass spectrometry (MS), and parallel reaction monitoring (PRM) analysis. In this study, we provide an MS–PRM data acquisition method for hFSH glycopeptides identification with high specificity and their quantification by extracting the chromatographic traces of selected fragments of glycopeptides. Once set up for all its features, the proposed method could be transferred to the clinic to improve fertility treatments and follow-ups in men and women.

Supramolecular Four-Armed Star A2B2 Copolymer (Miktoarm Star) via Host–Guest Complexation and Nitroxide-Mediated Radical Polymerization

A supramolecular four-armed star copolymer was prepared from the pseudorotaxane complex of a crown-centered polystyrene and a bis(2,2,6,6-tetramethylpiperidinyl-1-oxy) viologen compound and n-butyl methacrylate (BMA) by nitroxide-mediated polymerization of the latter. Solvent fractionation with dioxane/THF/MeOH removed a small amount of a high molecular weight poly(BMA) homopolymer. The molecular weight increase from its monomodal size exclusion chromatographic trace gives direct evidence of the four-armed star copolymer formation; there is no residual starting polystyrene. NMR spectroscopy shows the two components of the copolymer and confirms the presence of the central [2]rotaxane unit. Differential scanning calorimetry displays two glass-transition temperatures, which correspond closely to those of polystyrene and poly(BMA).

CHARACTERISTICS OF MILK FAT OF GREY UKRAINIAN CATTLE BREED

CHARACTERISTICS OF MILK FAT OF GREY UKRAINIAN CATTLE BREED

THE FATTY ACIDS CONTENT IN THE LIVER OF JAPANESE QUAILS AFTER THE CHEMICAL TREATMENT OF HATCHING EGGS

The five groups of Pharaoh quail (100 animals per group) were formed to fulfill the tasks. Quails of the control group were fed by the standard mixed fodder for quail (DSTU 4687:2006). The bird of the research groups (I–IV) received the same feed, but with the addition of 20 g/t of vitamin E. We selected the hatching eggs after 4 weeks of feeding research fodders. Eggs were weighed and laid for incubation using the standard mode after pre-incubation storage of the eggs of the quails obtained in the peak of egg production for 5 days. On the 14th day of incubation, the eggs of the quails were divided into 5 groups. Treatment of eggs I, II and III of the test group was carried out on the 14th day of incubation, respectively, with solutions of 1% sodium hypochlorite 2% perchloric acid 0,5% hydrogen peroxide. Egg of control and IV experimental group were not subject to chemical treatment. The material for research was hatching eggs of quail, liver tissue of 14 daily embryos and 1-day quail. The fatty acid composition of the lipids of tissues and egg yolk was determined in this biological material. Analysis of methyl esters of LC was carried out on a Gas chromatograph Trace GC Ultra (USA) with a flame ionization detector. Identification of fatty acids was carried out using a standard Supelco 37 Component FAME Mix. Quantitative assessment of the spectrum of fatty acids of yolk lipids was carried out by the method of internal normalization, determining their percentage content. Ontogenetic differences in the fatty acid composition of tissues are established. Processing of the eggshell on the fourteenth day of incubation with H2O2 solution is accompanied by an increase in the proportion of stearic acid in the liver of the 1-day quail and a decrease in the proportion of Neukosatrienoic and pre-fatty acids. Treatment of the shell with sodium hypochlorite and HCl is accompanied by an increase in the proportion of stearic acid by 0.96–1.00 % and аrachinic acid, with a decrease in the proportion of gondoinic and eicosatrienic fatty acids. At the same time, when treating with HCl and sodium hypochlorite, the ratio of the sum of saturated to unsaturated fatty acids decreases by 3.2–7.9% (p&lt;0.05). So, the established changes in the fatty acid composition of the liver one-day quail indicate a significant effect of the chemical treatment of the egg shell on the exchange of fatty acids in the embryonic period.

Адсорбция алифатических и циклических углеводородов С6 на Silochrome C-120, модифицированном наночастицами никеля и меди

В работе исследованы адсорбционные свойства материалов, содержащих наночастицы никеля и меди, полученных левитационно-струйным методом. Адсорбцию изучали методом обращенной газовой хроматографии. Адсорбатами служили шестичленные углеводороды, отличающиеся по количеству и характеру кратных связей, а также цикличностью или линейностью углеродного скелета.Были рассчитаны константы и теплоты адсорбции исследуемых углеводородов на адсорбенте, содер-жащем наночастицы никеля и меди; определены теплоты специфических взаимодействий.

Targeted Proteomics for Multiplexed Verification of Markers of Colorectal Tumorigenesis

Targeted proteomic methods can accelerate the verification of multiple tumor marker candidates in large series of patient samples. We utilized the targeted approach known as selected/multiple reaction monitoring (S/MRM) to verify potential protein markers of colorectal adenoma identified by our group in previous transcriptomic and quantitative shotgun proteomic studies of a large cohort of precancerous colorectal lesions. We developed SRM assays to reproducibly detect and quantify 25 (62.5%) of the 40 selected proteins in an independent series of precancerous and cancerous tissue samples (19 adenoma/normal mucosa pairs; 17 adenocarcinoma/normal mucosa pairs). Twenty-three proteins were significantly up-regulated (n = 17) or downregulated (n = 6) in adenomas and/or adenocarcinomas, as compared with normal mucosa (linear fold changes ≥ ±1.3, adjusted p value <0.05). Most changes were observed in both tumor types (up-regulation of ANP32A, ANXA3, SORD, LDHA, LCN2, NCL, S100A11, SERPINB5, CDV3, OLFM4, and REG4; downregulation of ARF6 and PGM5), and a five-protein biomarker signature distinguished neoplastic tissue from normal mucosa with a maximum area under the receiver operating curve greater than 0.83. Other changes were specific for adenomas (PPA1 and PPA2 up-regulation; KCTD12 downregulation) or adenocarcinoma (ANP32B, G6PD, RCN1, and SET up-regulation; downregulated AKR1B1, APEX1, and PPA1). Some changes significantly correlated with a few patient- or tumor-related phenotypes. Twenty-two (96%) of the 23 proteins have a potential to be released from the tumors into the bloodstream, and their detectability in plasma has been previously reported. The proteins identified in this study expand the pool of biomarker candidates that can be used to develop a standardized precolonoscopy blood test for the early detection of colorectal tumors.

ВМІСТ ЖИРНИХ КИСЛОТ В ЛІПІДАХ ФЕТАЛЬНИХ СТОВБУРОВИХ КЛІТИН КОТА

Defined content of fatty acids in lipids cat stem cells derived from fetal primary material. Fetal stem cells (FSCs) cat treated by culturing primary material in the CO2 incubator containing 5% CO2, at a temperature 37°С in DMEM medium with the addition of 15 – 20% fetal bulls serum and 1% antibiotic–antimycotic. When confluent monolayer reached 70 – 80%, the cells are removed from the culture dishes and held subcultivation to reduce the heterogeneity of culture. The resulting stem cells are tested for fatty acid content by gas–liquid chromatography.Determination of fatty acids in lipids fetal stem cells conducted under SOST ISO 5508–2001. Sample preparation was performed according to ISO 150 5509–2002 in our modification. A mixture of methyl esters of fatty acids were analyzed on the gas chromatograph Trace GC Ultra with flame ionization detector for capillary column SPTM –2560, 100 m x 0.25 mm ID, 0.20 μm film (Supelco). Identification of fatty acids was performed using a standard sample Supelco 37 Сomponent FAME Mix. Performed quantitative assessment by spectrum of crystal planes valuation peaks methylated derivatives LCD and determine their content as a percentage of the total content of all the LCD.Investigated that the lipids contained cat FSCs short–, medium– and long–chain fatty acids. In the lipid fetal stem cells found cat 18 Number fatty acids from saturated – most of palmitatе (34.53 ± 0.58%), with monounsaturated – оleate (20.20 ± 0.93%), with polyunsaturated – linoleic acid (6.27 ± 0.01%). Least composed of lipids of cells found cis–8.11.14–eykozatriyenovoyi acid (0.03 ± 0.01%).The total content of saturated fatty acids in the lipid cat FSCs is 67.75, unsaturated fatty acids – 32.25%. Saturation ratio is 2.10. Monoyenic fatty acids identified in the number of 23.19%, and polyenic – 9,06%. The index value n3 fatty acids to n6 in lipids cat FSCs is 0.35.

Dinosaur: A Refined Open-Source Peptide MS Feature Detector.

In bottom-up mass spectrometry (MS)-based proteomics, peptide isotopic and chromatographic traces (features) are frequently used for label-free quantification in data-dependent acquisition MS but can also be used for the improved identification of chimeric spectra or sample complexity characterization. Feature detection is difficult because of the high complexity of MS proteomics data from biological samples, which frequently causes features to intermingle. In addition, existing feature detection algorithms commonly suffer from compatibility issues, long computation times, or poor performance on high-resolution data. Because of these limitations, we developed a new tool, Dinosaur, with increased speed and versatility. Dinosaur has the functionality to sample algorithm computations through quality-control plots, which we call a plot trail. From the evaluation of this plot trail, we introduce several algorithmic improvements to further improve the robustness and performance of Dinosaur, with the detection of features for 98% of MS/MS identifications in a benchmark data set, and no other algorithm tested in this study passed 96% feature detection. We finally used Dinosaur to reimplement a published workflow for peptide identification in chimeric spectra, increasing chimeric identification from 26% to 32% over the standard workflow. Dinosaur is operating-system-independent and is freely available as open source on https://github.com/fickludd/dinosaur.

Bioinformatics tools for small genomes, such as hepatitis B virus.

DNA sequence analysis is undertaken in many biological research laboratories. The workflow consists of several steps involving the bioinformatic processing of biological data. We have developed a suite of web-based online bioinformatic tools to assist with processing, analysis and curation of DNA sequence data. Most of these tools are genome-agnostic, with two tools specifically designed for hepatitis B virus sequence data. Tools in the suite are able to process sequence data from Sanger sequencing, ultra-deep amplicon resequencing (pyrosequencing) and chromatograph (trace files), as appropriate. The tools are available online at no cost and are aimed at researchers without specialist technical computer knowledge. The tools can be accessed at http://hvdr.bioinf.wits.ac.za/SmallGenomeTools, and the source code is available online at https://github.com/DrTrevorBell/SmallGenomeTools.

A Comparison of the Metabolome of Male and Female Drosophila melanogaster

Metabolite profiles of ten batches of male and female Drosophila melanogaster were compared using three different chromatographic methods interfaced to an Orbitrap Exactive mass spectrometer. Several thousand features were observed and were reduced after data extraction and careful checking of around 390 metabolites excluding lipids. Chromatographic traces for these metabolites are presented as supplementary information. There were many significant differences between male and female flies. Female flies contained much higher levels of methylated lysines and methylated arginine suggesting differences in histone metabolism. In addition, there were differences in the methylation of nucleosides, and Sadenosylmethionine metabolism. Differences in the methylome may relate to the requirement of compensation for the single X chromosome present in males since methylated histones inhibit gene transcription. Nucleoside phosphate levels were elevated in female flies which may relate to increased requirement for DNA biosynthesis for egg production. A series of acylated amino acids previously observed in Drosophila was further characterised and these metabolites were present to a much greater extent in female flies and may be associated with the microbiome. Keywords: Drosophila melanogaster, male female metabolomics, methylation, acylated amino acids.

Influence of Polyethylene Pipe on the Quality of Water in a Water Distribution System

In the past two decades economic considerations and very good mechanical parameters of polyethylene pipes contributed to more and more common use of the pipes for construction and repair of water distribution systems all over world. This paper presents the results of research into parameters of water collected from a polyethylene pipeline which was a part of an operational water supply network. We determined Total Organic Carbon (TOC) content and the content of organic compounds in the examined water samples using a multichannel gas chromatograph (Trace Ultra Thermo) coupled with a mass spectrometer (Polaris Q). We identified organic compounds bounded with antioxidants, added to polyethylene in the process of pipe production, in the water samples.

Phylogenetic diversity of dominant bacterial communities during bioremediation of crude oil-polluted soil

Bioremediation of hydrocarbon pollutants is advantageous owing to the costeffectiveness of the technology and the ubiquity of hydrocarbon degrading microorganisms in the soil. Soil microbial diversity is affected by hydrocarbon perturbation thus selective enrichment of hydrocarbon utilizers occurs. Hydrocarbons interact with the soil matrix and soil microorganisms determining the fate of the contaminants relative to their chemical nature and microbial degradative capabilities respectively. Bacterial dynamics in crude oil-polluted soil microcosms undergoing bioremediation were investigated over a 42-day period. Four out of the five microcosms containing 4kg of pristine soil each were contaminated with 4% Arabian light crude oil. Three microcosms were amended with either 25g of NPK fertilizer, calcium ammonium nitrate or poultry droppings respectively while the fourth designated oilcontaminated control was unamended. The fifth microcosm had only pristine soil and was set up to ascertain indigenous bacterial community structure pre-contamination. Biostimulated soils were periodically tilled and watered. Hydrocarbon degradation was measured throughout the experimental period by gas chromatography. Gas chromatographic tracing of residual hydrocarbons in biostimulated soils showed marked attenuation of contaminants starting from the second (day 14) till the sixth (day 42) week after contamination whereas no significant reduction in hydrocarbon peaks was seen in the oil contaminated control soil throughout the 6-week experimental period. Molecular fingerprints of bacterial communities involved in aerobic biodegradation of crude oil hydrocarbons in biostimulated soils and controls were generated with DGGE using PCR-amplification of 16S rRNA gene obtained from extracted total soil community DNA. DGGE fingerprints demonstrated that NPK, calcium ammonium nitrate and poultry droppings selected different bacterial populations during the active phase of oil degradation. Cluster analysis of DGGE bands using simple matching group average setting revealed that poultry droppings-amended soils and calcium ammonium nitrateamended soils formed distinct clades meaning that the treatment selected similar bacterial populations for each of the treatments whereas NPK soils showed less association. Excision, reamplification and sequencing of dominant DGGE bands in biostimulated soils revealed the presence of distinct hydrocarbon degraders like Corynebacterium spp., Dietzia spp., low G+C Gram positive bacteria and some uncultured bacterial clones. Phylogenetic analysis of the 16S rRNA gene sequences of these dominant bacterial communities was conducted using the neighbour joining method of PHYLIP. Two distinct clades appeared in the tree clustered members of the Actinobacteria and Firmicutes separately. The overall data suggested that Gram positive bacteria especially members of the Actinobacteria may have a key role in bioremediation of crude oil-polluted soil.

Combined cation-exchange and extraction chromatographic method of pre-concentration and concomitant separation of Cu(II) with high molecular mass liquid cation exchanger after its online detection

A selective method has been developed for the extraction chromatographic trace level separation of Cu(II) with Versatic 10 (liquid cation exchanger) coated on silanised silica gel (SSG-V10). Cu(II) has been extracted from 0.1 M acetate buffer at the range of pH 4.0–5.5. The effects of foreign ions, pH, flow-rate, stripping agents on extraction and elution have been investigated. Exchange capacity of the prepared exchanger at different temperatures with respect to Cu(II) has been determined. The extraction equilibrium constant ( K ex) and different standard thermodynamic parameters have also been calculated by temperature variation method. Positive value of Δ H (7.98 kJ mol −1) and Δ S (0.1916 kJ mol −1) and negative value of Δ G (−49.16 kJ mol −1) indicated that the process was endothermic, entropy gaining and spontaneous. Preconcentration factor was optimized at 74.7 ± 0.2 and the desorption constants K desorption 1(1.4 × 10 −2) and K desorption 2(9.8 × 10 −2) were determined. The effect of pH on R f values in ion exchange paper chromatography has been investigated. In order to investigate the sorption isotherm, two equilibrium models, the Freundlich and Langmuir isotherms, were analyzed. Cu(II) has been separated from synthetic binary and multi-component mixtures containing various metal ions associated with it in ores and alloy samples. The method effectively permits sequential separation of Cu(II) from synthetic quaternary mixture containing its congeners Bi(III), Sn(II), Hg(II) and Cu(II), Cd(II), Pb(II) of same analytical group. The method was found effective for the selective detection, removal and recovery of Cu(II) from industrial waste and standard alloy samples following its preconcentration on the column. A plausible mechanism for the extraction of Cu(II) has been suggested.

The effect of carbohydrate additives in the freeze-drying of alkaline phosphatase.

Alkaline phosphatase was used as a model in studies to assess the effects of lyophilization on biological activity and molecular integrity in the presence or absence of added carbohydrate. The stability of the activity of alkaline phosphatase, lyophilized in Tris buffer alone or in the presence of the carbohydrates mannitol, lactose or trehalose was examined. Enzyme activity in formulations with Tris buffer alone or with mannitol was considerably reduced by freeze-drying and further storage at elevated temperatures; freeze-drying with mannitol failed to maintain activity at a temperature of 37 degrees C over 21 days, whilst the loss of activity was more gradual when freeze-dried in buffer alone and stored at higher temperatures. Lactose and trehalose maintained the alkaline phosphatase activity after freeze-drying and, furthermore, preparations containing trehalose retained activity even when the material was subjected to temperatures of up to 45 degrees C for up to 84 days. At 56 degrees C the alkaline phosphatase activity did not show a significant drop until 14 days with the lactose formulation or until 21 days with trehalose. After 84 days at 56 degrees C, 30% of the activity still remained in the formulation containing trehalose. In addition to the changes in the enzyme activity, FPLC chromatographic traces and SDS-PAGE gels demonstrated compositional differences between each formulation after storage.

Effects of cyclodextrins on the acid hydrolysis of digoxin

The effects of three cyclodextrins (alpha-, beta-, gamma-CyD) on the acid hydrolysis of digoxin were examined. From the high performance liquid chromatographic tracing of each of the four components (digoxin, bisdigitoxoside, monodigitoxoside, digoxigenin) in reaction mixtures, the individual rate constants (K1-K6) were determined by analogue computer simulation. The hydrolysis was suppressed by CyDs in the order of beta-great than gamma-greater than alpha-greater than-CyD, where beta-CyD inhibited the appearance rates of digoxigenin (k3, K5, and K6) significantly. In the dissolution study of digoxin tablets, the increase in dissolution rate and decrease in acid hydrolysis were attained by inclusion complexation. The data are presented suggesting that CyDs are useful for improving the oral bioavailability of digoxin.

Speciation of Iodine in High Iodine Groundwater in China Associated with Goitre and Hypothyroidism

Iodine intake affects the occurrence of disease in a population. Excessive iodine intake may be caused by a high iodine content of drinking water. Tap water in few locations in Europe contains up to 139 microg/L mostly bound to humic substances, probably leaching from marine sediments in the aquifers. Even higher iodine contents have been found in Chinese waters, previously shown to associate with goitre and hypothyroidism. The aims were to elucidate speciation of high iodine groundwater from deep wells in China and to compare with high iodine waters from Europe. Water was sampled from eight wells in five villages along Bohai Bay, China. Macro-molecules and low molecular weight (MW) substances were separated by size exclusion chromatography (high performance liquid chromatography, Superose 12 HR 10/30, buffer 0.1 M Tris, pH 7.0). Organic material was evaluated by A280 and iodine in fractions measured by the Ce/As method after alkaline incineration. Iodine content of well water varied from 135 to 880 microg/L (median 287 microg/L). The amount of organic material in water was low with A280, <1-5 mAU. The chromatographic traces were similar between samples: One peak of iodine eluted around K (AV) 0.65 corresponding to MW 5 kDa (humic substances) and one peak at V (total) (iodide/low MW substances). The fraction of iodine in macro-molecules, suggested to be humic substances, varied from 8% to 70% (median 27%). Iodine and peak absorbance were associated (p = 0.006). In conclusion, iodine in iodine-rich deep well water in northern China may have marine origin and may associate with humic substances, comparable to shallow well iodine-rich water in Europe. High iodine intake from iodine-rich water suggests the cause of endemic goitre and hypothyroidism in some areas in China being iodine.

Analysis of biologically active compounds in water by ultra‐performance liquid chromatography quadrupole time‐of‐flight mass spectrometry

A new method based on ultra-performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry ((Q-ToF)-MS) was developed for the analysis of 32 biologically active compounds including anti-inflammatories, analgesics, lipid regulators, psychiatric drugs, anti-ulcer agents, antibiotics, beta-blockers and phytoestrogens. This new method allows chromatographic analysis in 14 min, with instrumental detection limits from 2 to 84 pg, and limits of quantification ranging from 0.1 to 15 ng/L in tap water, and from 2 to 300 ng/L in wastewater. The potential of liquid chromatography with triple quadrupole mass spectrometry (LC/QqQ-MS) was compared with that of UPLC/(Q-ToF)-MS for the analysis of biologically active compounds in water samples. LC/Q-ToF provides accurate mass information and a significantly higher mass resolution than quadrupole analyzers. The available mass resolution of ToF instruments diminishes the problem of isobaric interferences and helps the analysis of trace compounds in complex samples. In this work UPLC/Q-ToF chromatograms were recorded containing full scan spectral data. The m/z values of analytes were extracted from the total ion chromatogram (TIC) and the accurate masses of the compounds were obtained. In addition, to increase the selectivity of ToF measurements a narrow accurate mass interval (20 m m/z units mass window) was used to reconstruct the chromatographic traces. However, regarding quantitative performance in terms of dynamic range and limits of detection (LODs), typical LODs achieved by QqQ instruments operating in multiple-reaction monitoring (MRM) mode ranged from 1 to 50 ng/L in wastewater, and the linear response for QqQ instruments generally covers three orders of magnitude. This is an important advantage over ToF instruments and one of the reasons why QqQ instruments are widely used in quantitative environmental analysis.