Uptake, translocation, and metabolization of amitriptyline, lidocaine, orphenadrine, and tramadol by cress and pea.

The uptake, translocation, and metabolization of four widely used drugs, amitriptyline, orphenadrine, lidocaine, and tramadol, were investigated in a laboratory study. Cress (Lepidium sativum L.) and pea (Pisum sativum L.) were employed as model plants. These plants were grown in tap water containing the selected pharmaceuticals at concentrations ranging from 0.010 to 10 mg L-1, whereby the latter concentration was employed for the (tentative) identification of drug-related metabolites formed within the plant. Thereby, mainly phase I metabolites were detected. Time-resolved uptake studies, with sampling after 1, 2, 4, 8, and 16 days, revealed that all four pharmaceuticals were taken up by the roots and further relocated to plant stem and leaves. Also in these studies, the corresponding phase I metabolites could be detected, and their translocation from root to stem (pea only) and finally leaves could be investigated.

Uptake and metabolization of four sartan drugs by eight different plants: Targeted and untargeted analyses by HPLC-drift-tube-ion-mobility quadrupole time-of-flight mass spectrometry.

In this study, we investigated the uptake and metabolization of four drugs (plus the associated prodrugs) from the sartan family by eight edible plants. Growing the plants hydroponically in a medium containing the respective drug, more than 40 phases I and II metabolites derived from the four sartan drugs could be tentatively identified. To demonstrate the suitability of the proposed analytical approach for actual environmental samples, garden cress (Lepidium sativum) selected as a model plant was grown in water drawn from the effluent of two local wastewater treatment plants. Thereby, three of the sartans, namely, olmesartan, candesartan, and valsartan, could be found in the plant extracts at concentrations of 3.1, 10.4, and 14.4 ng g-1 , respectively. Additionally, for candesartan and valsartan, a glycosylated transformation product could be detected. In order to extend the present (targeted) workflow also toward the analysis of unknown transformation products (i.e., those not listed in the custom-made database used for this research), a nontargeted approach for the analysis of plant extracts with respect to the presence of drug-related metabolites was developed. Comparison of the targeted and the nontargeted workflows led to the finding of two additional, so far unidentified, transformation products originating from azilsartan.

Uptake and bio-transformation of telmisartan by cress (Lepidium sativum) from sewage treatment plant effluents using high-performance liquid chromatography/drift-tube ion-mobility quadrupole time-of-flight mass spectrometry.

In the present study, the uptake and metabolization of the sartan drug telmisartan by a series of plants was investigated. Thereby for seven potential metabolites, modifications on the telmisartan molecule such as hydroxylation and/or glycosylation could be tentatively identified. For two additional signals detected at accurate masses m/z 777.3107 and m/z 793.3096, no suggestions for molecular formulas could be made. Further investigations employing garden cress (Lepidium sativum) as a model plant were conducted. This was done in order to develop an analytical method allowing the detection of these substances also under environmentally relevant conditions. For this reason, the knowledge achieved from treatment of the plants with rather high concentrations of the parent drug (10 mg L-1) was compared with results obtained when using solutions containing telmisartan in the µg - ng L-1 range. Thereby the parent drug and up to three tentative drug-related metabolites could still be detected. Finally cress was cultivated in water taken from a local waste water treatment plant effluent containing 90 ng L-1 of telmisartan and harvested and the cress roots were extracted. In this extract, next to the parent drug one major metabolite, namely telmisartan-glucose could be identified.

Time study on the uptake of four different beta-blockers in garden cress (Lepidium sativum) as a model plant.

The aim of this study was to investigate the uptake of four beta-blockers by the model plant Lepidium sativum (garden cress) and their possible metabolization over a time period of 8 days. Therefore, cress was grown hydroponically in tap water for a week until they were matured, following irrigation with drug-containing water over the course of another 8 days. Samples were taken at days 1, 2, 4, and 8 after irrigation started. All four beta-blockers were taken up by the plants and the different octanol-water coefficients (log P) of the drugs have an influence on the uptake speed in the roots of the plants. The log P seems to have no influence on the translocation of the drugs from the root to the shoots. Furthermore, neither phase I nor phase II metabolization occurred inside the plants.

A fast-screening approach for the tentative identification of drug-related metabolites from three non-steroidal anti-inflammatory drugs in hydroponically grown edible plants by HPLC-drift-tube-ion-mobility quadrupole time-of-flight mass spectrometry.

The (tentative) identification of unknown drug-related phase II metabolites in plants upon drug uptake remains a challenging task despite improved analytical instrument performance. To broaden the knowledge of possible drug metabolization, a fast-screening approach for the tentative identification of drug-related phase II metabolites is presented in this work. Therefore, an in silico database for the three non-steroidal anti-inflammatory drugs (ketoprofen, mefenamic acid, and naproxen) and a sub-group of their theoretical phase II metabolites (based on combinations with glucose, glucuronic acid, and malonic acid) was created. Next, the theoretical exact masses (protonated species and ammonia adducts) were calculated and used as precursor ions in an autoMS/MS measurement method. The applicability of this workflow was tested on the example of eleven edible plants, which were hydroponically grown in solutions containing the respective drug at a concentration level of 20 mg/L. For the three drugs investigated this led to the tentative identification of 41 metabolites (some of them so far not described in this context), such as combinations of hydroxylated mefenamic acid with up to four glucose units or hydroxylated mefenamic acid with two glucose and three malonic acid units.

A new analytical workflow using HPLC with drift-tube ion-mobility quadrupole time-of-flight/mass spectrometry for the detection of drug-related metabolites in plants.

Investigations into the interaction of xenobiotics with plants (and in particular edible plants) have gained substantial interest, as water scarcity due to climate-change-related droughts requires the more frequent use of reclaimed wastewaters for irrigation in agriculture. Non-steroidal anti-inflammatory drugs are common contaminants found in wastewater treatment plant effluents. For this reason, the interaction of nine edible plants with diclofenac (DCF), a widely used representative of this group of drugs, was investigated. For this purpose, plants were hydroponically grown in a medium containing DCF. For the detection of unknown DCF-related metabolites formed in the plant upon uptake of the parent drug' a new workflow based on the use of HPLC coupled to drift-tube ion-mobility quadrupole time-of-flight/mass spectrometry (DTIM QTOF-MS) was developed. Thereby' for chromatographic peaks eluting from the HPLC, drift times were recorded, and analytes were subsequently fragmented in the DTIM QTOF-MS to provide significant fragments. All information available (retention times, drift times, fragment spectra, accurate mass) was finally combined' allowing the suggestion of molecular formulas for 30 DCF-related metabolites formed in the plant, whereby 23 of them were not yet known from the literature.

Investigations on the uptake and transformation of sunscreen ingredients in duckweed (Lemna gibba) and Cyperus alternifolius using high-performance liquid chromatography drift-tube ion-mobility quadrupole time-of-flight mass spectrometry.

The uptake, translocation and transformation of three UV-blockers commonly employed in sunscreens, namely avobenzone, octocrylene and octisalate from water by Lemna gibba and Cyperus alternifolius was investigated. Reversed phase high performance liquid chromatography coupled to drift-tube ion-mobility quadrupole time-of-flight mass spectrometry was used for analyzing the extracts from the selected plants after incubation with the UV-blockers for one week. For avobenzone several transformation products resulting from hydroxylation, demethylation and oxidation of the parent molecule could be identified by measuring accurate mass, performing MS/MS experiments and by determining their drift-tube collision cross sections employing nitrogen as drift gas. In addition, the plants were subjected to two commercially available sunscreens, providing similar results to those obtained for the standard solutions of the UV-blockers. Finally, a kinetic study on the uptake and transformation of avobenzone, octocrylene and octisalate was conducted over a period of 216 h, revealing that the UV-filters were mostly present in their parent form and only to a smaller part converted into transformation products.

High-performance liquid chromatography drift-tube ion-mobility quadrupole time-of-flight/mass spectrometry for the identity confirmation and characterization of metabolites from three statins (lipid-lowering drugs) in the model plant cress (Lepidium sativum) after uptake from water.

This work describes the metabolization of three different statins (lipid-lowering drugs) namely Atorvastatin, Fluvastatin and Simvastatin in the model plant cress (Lepidium sativum) after uptake from the growing medium. Analyzing plant extracts with HPLC hyphenated with a drift-tube ion-mobility quadrupole time-of-flight / mass spectrometer allowed the identity confirmation of more than 45 metabolites, resulting from oxidation/dehydrogenation, dehydration or hydroxylation of the parent drug or conjugation with amino acids and sugars. Metabolites were characterized by their retention times, m/z ratios, fragmentation patterns in MS/MS experiments, and their collision cross sections. Furthermore, a targeted analysis method for the trace-level analysis of the parent drugs as well as their metabolites in plant extracts was implemented by using HPLC coupled to triple quadrupole mass spectrometry in the multiple reaction monitoring mode. This approach was applied to study the metabolite distribution within the plant and to detect relative changes in metabolite concentrations as a function of growth time. Using a modified QuEChERS approach for extraction more than 50% of the metabolites could be still detected, if plants were exposed to only 1-10 µg L-1 of each statin.

Uptake and metabolism of the antidepressants sertraline, clomipramine, and trazodone in a garden cress (Lepidium sativum) model.

Environmental contamination with pharmaceuticals has received growing attention in recent years. Several studies describe the presence of traces of drugs in water bodies and soils and their impacts on nontarget organisms including plants. Due to these facts investigations of the uptake and metabolism of pharmaceuticals in organisms is an emerging research area. The present study demonstrates the analysis of three selected antidepressants (sertraline, clomipramine, and trazodone) as well as metabolites and transformation products in a cress model (Lepidium sativum). Cress was treated with tap water containing 10 mg/L of the parent drugs. Employing an analytical approach based on high performance liquid chromatography coupled with quadrupole time of flight or Orbitrap mass spectrometry in MS and MS² modes, in total 14 substances were identified in the cress extracts. All three parent drugs were taken up by the cress and translocated from the roots to the leaves in specific patterns. In addition to this, eleven metabolite species were identified. They were generated by hydroxylation, demethylation, conjugation with amino acids, or combinations of these mechanisms. Finally, the inclusion of control cultures in the experimental setup allowed for a differentiation of "true" metabolites generated by the cress and transformation products generated by plant-independent mechanisms.

Insights into the uptake, metabolization, and translocation of four non-steroidal anti-inflammatory drugs in cress (Lepidium sativum) by HPLC-MS2.

The metabolization of four non-steroidal anti-inflammatory drugs by cress (Lepidium sativum) was investigated using a HPLC-MS2 method. Cress was grown hydroponically in water containing 0.1 mg/L of each drug for investigations on the kinetics of drug uptake and metabolization over a growing period of 12 days. It could be shown that the parent drugs are metabolized and the abundance of both the parent drug and the metabolites formed, varies over time. Furthermore the distribution of the investigated substances within the different plant parts changed throughout the duration of the experiment due to translocation. Finally cress was cultivated in a solution containing the four drugs in concentrations as low as 0.001 mg/L to resemble the situation in real reclaimed wastewaters. Employing a QuEChERS approach for sample extraction and HPLC-MS2 in the multiple reaction monitoring mode allowed detecting nine metabolites in this cress sample.

High-performance liquid chromatography - mass spectrometry analysis of the parent drugs and their metabolites in extracts from cress (Lepidium sativum) grown hydroponically in water containing four non-steroidal anti-inflammatory drugs.

In this paper the metabolism of four non-steroidal anti-inflammatory drugs, (ketoprofen, mefenamic acid, naproxen, and diclofenac) by cress (Lepidium sativum) is described. Cress was cultivated hydroponically in water spiked with the parent drugs at levels ranging from 0.01mgL-1 to 1mgL-1. Employing an approach based on the analysis of the plant extracts by HPLC coupled either with quadrupole-time-of-flight mass spectrometry, or Orbitrap MS or triple quadrupole (QqQ) MS allowed the identification of twenty substances (sixteen metabolites and four parent drugs). Metabolites were formed from the parent drug by hydroxylation or conjugation with polar molecules such as glucose, small organic acids or amino acids. Introducing a pre-concentration step employing solid-phase extraction and using HPLC-QqQ/MS in the multiple reaction monitoring mode enabled the positive detection of 11 of the proposed metabolites next to the four parent components even in plants grown in a 0.01mgL-1 solution of the tested drugs, which is close to the conditions in real reclaimed waters.

Nonaqueous Capillary Electrophoresis Mass Spectrometry.

The term nonaqueous capillary electrophoresis (NACE) commonly refers to capillary electrophoresis with purely nonaqueous background electrolytes (BGE). Main advantages of NACE are the possibility to analyze substances with very low solubility in aqueous media as well as separation selectivity that can be quite different in organic solvents (compared to water)-a property that can be employed for manipulation of separation selectivities. Mass spectrometry (MS) has become more and more popular as a detector in CE a fact that applies also for NACE. In the present chapter, the development of NACE-MS since 2004 is reviewed. Relevant parameters like composition of BGE and its influence on separation and detection in NACE as well as sheath liquid for NACE-MS are discussed. Finally, an overview of the papers published in the field of NACE-MS between 2004 and 2014 is given. Applications are grouped according to the field (analysis of natural products, biomedical analysis, food analysis, analysis of industrial products, and fundamental investigations).

Advances in the determination of hindered amine light stabilizers - A review.

Within this paper we discuss analytical strategies for the characterization and quantitation of hindered amine light stabilizers (HALS) an important sub-group of polymer additives. For the determination of monomeric HALS a range of mature and reliable techniques exists, allowing their determination in polymer extracts. If qualitative or semi-quantitative information suffices, certain techniques are capable of sampling directly from the polymer surface with limited or no sample preparation. Different strategies for the determination of complex oligomeric HALS in extracts from polymer samples are discussed. Here, approaches providing only a sum parameter including all HALS oligomers have been distinguished from more sophisticated technologies allowing the determination of single oligomers, their degradation and by-products. Particularly, the latter issue is facing increased interest as it provides important information for polymers aging studies. A tabulated overview provides comprehensive information on different analytical techniques suitable for HALS determination.

Analysis of saccharides in beverages by HPLC with direct UV detection.

The present study demonstrates the suitability of direct UV detection for saccharide analysis in HPLC. Under highly alkaline conditions, the non-UV absorbing saccharides are converted by a photo-initiated chemical reaction in the detection cell into malonenolate, which can be detected at 266 nm. A straightforward method for such direct UV detection of saccharides after their separation by anion-exchange chromatography was developed and successfully applied to several beverage samples. Investigation and optimization of the influencing factors using design of experiment resulted in a baseline separation of glucose, fructose, and sucrose within 6 min and LOD values below 0.2 mg L(-1). In addition, a fast, simple and cost-effective flow injection method was developed to estimate the total saccharide concentration. The results of this method applied to beverage samples are in good agreement with the chromatographic method as well as to the saccharide concentration stated by the manufacturer. Finally, a comparison of different commercially available UV detectors and detector cells revealed that sensitive detection requires the use of recently introduced flow cells with extended path length.

Direct ionization methods in mass spectrometry: An overview.

Within this paper a sub-group of ambient ionization mass spectrometry namely direct ionization mass spectrometry techniques are reviewed. They are characterized by the generation of an electrospray directly from the sample investigated. Prominent representatives include paper spray mass spectrometry, tissue spray mass spectrometry, probe electrospray ionization or thin-layer chromatography mass spectrometry. Applications of all major direct ionization techniques within different fields such as biomedical analysis, analysis of natural products, analysis of technical products and food analysis, just to name a few, are discussed and relevant parameters are listed in five Tables.

Identification and semi-quantitative determination of anti-oxidants in lubricants employing thin-layer chromatography-spray mass spectrometry.

A quick and simple method for identification and semi-quantitative determination of nine antioxidants commonly used in lubricants is presented. A dual step thin-layer chromatography (TLC) separation, removes in a first step the oil matrix whereas in a second step the antioxidants are separated. Cutting the spots out of the TLC-plate in the form of triangles allows direct-spray mass spectrometric (MS) measurements, providing MS and MS(n) spectra (if an appropriate MS instrument is employed) of the antioxidants, allowing their identification but also giving information about potential oxidation or degradation of these additives. Calibration curves within the concentration range relevant for the analysis of real oil samples (0.2-1.2gL(-1)) were constructed with R(2) values above 0.98 (when using an appropriate internal standard). This allowed the semi-quantitative determination of the selected antioxidants in real oils samples. Comparison with results from HPLC-UV measurement showed acceptable agreement for all analytes.

Using sheath-liquid reagents for capillary electrophoresis-mass spectrometry: application to the analysis of phenolic plant extracts.

The combination of CE and MS is now a widely used tool that can provide a combination of high resolution separations with detailed structural information. Recently, we highlighted the benefits of an approach to add further functionality to this well-established hyphenated technique, namely the possibility to perform chemical reactions within the sheath-liquid of the CE-MS interface . Apart from using hydrogen/deuterium exchange for online determination of numbers of exchangeable protons, the addition of DPPH• (2,2-diphenyl-1-picrylhydrazyl) to the sheath-liquid can be used as a fast screening tool for studying antioxidant characteristics of individual components. Such a CE-MS methodology allows rapid and information-rich analysis with minimal reagent and sample consumption to be performed. In the present work, we demonstrate the applicability of this approach for the characterization of phenolic plant extracts from the Labiatae family, namely Rosmarinus officinalis and Melissa officinalis. Using the described approach, a wide range of compounds (15 and 13 phenolic compounds, respectively) could be confidently identified using a combination of high resolution CE-MS separations with implementation of online deuterium exchange and DPPH• reactions. These compounds included polyphenols, phenolic acids, and triterpene acids. In conjunction with online MS/MS experiments, extensive structural information for aglyconic and glycosylated antioxidants present in the extracts could be obtained using simple experimental changes, which can be carried out prior to the purchasing of expensive chemical standards or the time-consuming preparative isolation of individual compounds.