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.

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.

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 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.

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.

Drift-Tube Ion Mobility-Mass Spectrometry for Nontargeted 'Omics.

This chapter describes the developments in drift-tube ion mobility-mass spectrometry (DTIM-MS) that have driven application development in 'omics analyses. Harnessing the additional, orthogonal separation that DTIM provides increased confidence in compound identifications as the mass spectral complexity can be reduced and mobility-derived parameters (most prominently the collision cross section, CCS) used to support identity confirmation goals for a variety of 'omics application areas. Presented within this contribution is a methodology for improving the transmission and maintaining accurate determination of drift time-derived CCS (DTCCS) for low molecular weight compounds for a typical nontargeted 'omics (metabolomics) workflow using liquid chromatography in combination with DTIM-MS.

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.

Conditions for Analysis of Native Protein Structures Using Uniform Field Drift Tube Ion Mobility Mass Spectrometry and Characterization of Stable Calibrants for TWIM-MS.

Determination of collisional cross sections (CCS) by travelling wave ion mobility mass spectrometry (TWIM-MS) requires calibration against standards for which the CCS has been measured previously by drift tube ion mobility mass spectrometry (DTIM-MS). The different extents of collisional activation in TWIM-MS and DTIM-MS can give rise to discrepancies in the CCS of calibrants across the two platforms. Furthermore, the conditions required to ionize and transmit large, folded proteins and assemblies may variably affect the structure of the calibrants and analytes. Stable hetero-oligomeric phospholipase A2 (PDx) and its subunits were characterized as calibrants for TWIM-MS. Conditions for acquisition of native-like TWIM (Synapt G1 HDMS) and DTIM (Agilent 6560 IM-Q-TOF) mass spectra were optimized to ensure the spectra exhibited similar charge state distributions. CCS measurements (DTIM-MS) for ubiquitin, cytochrome c, holo-myoglobin, serum albumin and glutamate dehydrogenase were in good agreement with other recent results determined using this and other DTIM-MS instruments. PDx and its ß and γ subunits were stable across a wide range of cone and trap voltages in TWIM-MS and were stable in the presence of organic solvents. The CCS of PDx and its subunits were determined by DTIM-MS and were used as calibrants in determination of CCS of native-like cytochrome c, holo-myoglobin, carbonic anhydrase, serum albumin and haemoglobin in TWIM-MS. The CCS values were in good agreement with those measured by DTIM-MS where available. These experiments demonstrate conditions for analysis of native-like proteins using a commercially available DTIM-MS instrument, characterize robust calibrants for TWIM-MS, and present CCS values determined by DTIM-MS and TWIM-MS for native proteins to add to the current literature database. Graphical Abstract ᅟ.