Dataset Information

GNPS - Urine MS/MS data used for Antihypertensive Drug Screening study

ABSTRACT: Urine MS/MS data acquired to test Molecular Networking as tool for drug screening in human urine to assist in drug adherence monitoring. Samples from a antihypertensive drug adherence human cohort were retroperspectively selected to test how many commonly prescribed antihypertensives and other over-the-counter drugs were picked up by Molecular Networking in 26 extracts. In addition, several endogenous molecular families were also annotated. Data was obtained in positive and in negative ionization mode on a Thermo Scientific Q-Exactive Orbitrap instrument, using stepped collision energy HCD fragmentation in a data-dependent-manner (DDA). Urine samples from anonymized human volunteers were used from a clinical sample set in the Glasgow Polyomics archive. These samples were obtained as part of a trial for which ethical approval was applied for through the Multi-centre Research and Ethics Committee (MREC), which was granted by the Scottish MREC and (with MREC N°06/MRE00/106). Informed consent was obtained from all individual study participants. Spot urine samples were obtained from the cohort of elderly hypertensive patients upon their first admission in the clinic. Urine extracts of 26 patients were selected as follows: diagnosed with hypertension, taking in a variety of different antihypertensive drugs (i.e., different drug classes), and availability of the sample extract in the Glasgow Polyomics archive. The resulting subject’s age range spanned from 42 to 87; 15 were male, 11 female; 4 were smokers; 5 were reported to have diabetes; and each reportedly took from 2 to 7 different classes of antihypertensive drugs. Full details also on data acquisition and analysis can be found in Van der Hooft et al. 2016, Metabolomics. Chromatography and Mass Spectrometry details: The samples were analysed using a Thermo Scientific Ultimate 3000 RSLCnano system (Thermo Scientific, CA, USA). The pHILIC separation was performed with a SeQuant ZIC-pHILIC column (150 x 4.6 mm, 5 microm) equipped with the corresponding pre-column (Merck KGaA, Darmstadt, Germany) - the column temperature was maintained at 25 C. A linear biphasic LC gradient was conducted from 80% B to 20% B over 15 min, followed by a 2 min wash with 5% B, and 8 min re-equilibration with 80% B, where solvent B is acetonitrile and solvent A is 20 mM ammonium carbonate in water. The flow rate was 300 microL/min, column temperature was held at 25 C, injection volume was 10 microL, and samples were maintained at 4 C in the autosampler. MS equipment and settings used: The LC system was coupled to a Thermo Scientific Q-Exactive Orbitrap mass spectrometer equipped with a HESI II interface (Thermo Scientific, Hemel Hempstead, UK). The set up was calibrated (Thermo calmix) in both ionization modes and tuned for the lower m/z range before analysis. Full scan (MS1) data was acquired in positive and negative switching mode in profile mode at 35,000 resolution (at m/z 200) using 1 microscan, an AGC target of 106 cts, a maximum injection time of 250 ms, with spray voltages +3.8 and -3.0 kV, capillary temperature 320 C, heater temperature of 150 C, sheath gas flow rate 40 a.u., auxiliary gas flow rate 5 a.u., sweep gas flow rate 5 a.u, a full scan mass window of 70-1050 m/z, and using m/z 74.0964 (+) and m/z 112.98563 (-) as locking masses. Fragmentation data (LC-MS/MS) was obtained in positive and negative ionization combined and separate fragmentation modes as described in (van der Hooft et al 2016). Briefly, for separate mode, a duty cycle consisted of one full scan (MS1) event and one Top5 (or Top10) MS/MS (MS2) fragmentation event, with full scan (MS1) resolution (at m/z 200) was set to 70,000, the AGC target set to 1 x 106, and the maximum injection time set to 120 ms. MS/MS (MS2) resolution (at m/z 200) was set to 17,500, the AGC target set to 2 x 105, MS/MS maximum injection time was set to 80 ms and the underfill ratio was set to 10 %, with a resulting intensity threshold of 2.5 × 105 cts. For combined mode, a duty cycle consisted of two of the above events in positive and negative ionization mode with the following modifications: full scan (MS1) resolution (at m/z 200) was set to 35,000, MS/MS resolution was set to 35,000, the AGC target was set to 1 x 105, and the maximum MS/MS filling time was set to 120 ms with an underfill ratio of 20%, resulting in an intensity threshold of 1.7 x 105. Further settings were as specified for full scan analysis above.

OTHER RELATED OMICS DATASETS IN: MSV000083528MTBLS307

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Justin van der Hooft

PROVIDER: MSV000082971 | GNPS |

REPOSITORIES: GNPS

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Urinary antihypertensive drug metabolite screening using molecular networking coupled to high-resolution mass spectrometry fragmentation.

van der Hooft Justin J J JJ Padmanabhan Sandosh S Burgess Karl E V KE Barrett Michael P MP

Metabolomics : Official journal of the Metabolomic Society 20160705

<h4>Introduction</h4>Mass spectrometry is the current technique of choice in studying drug metabolism. High-resolution mass spectrometry in combination with MS/MS gas-phase experiments has the potential to contribute to rapid advances in this field. However, the data emerging from such fragmentation spectral files pose challenges to downstream analysis, given their complexity and size.<h4>Objectives</h4>This study aims to detect and visualize antihypertensive drug metabolites in untargeted metab ...[more]

PMID: 27471437

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Project description:This dataset contains LC-MS/MS data in positive ionisation mode of 100 spot urine samples and a pooled sample thereof that were obtained in Glasgow, UK (Institute of Cardiovascular and Medical Sciences & Glasgow Polyomics, University of Glasgow) as part the 'Next Generation Sequencing and Metabolomic Approaches in Stratification of Resistant Hypertension' study for which ethical approval was granted (NHS Ethics Approval Number -14/LO/188). Urine samples have been collected under a range of controlled settings. Informed consent was obtained from all individual study participants. Spot urine samples were taken around 11 a.m. upon a visit to the clinic, 20 1.5 ml aliquots were taken and subsequently centrifuged at 1000 g for 10 minutes after which the supernatant was stored at -80 Celcius until further analysis. A general metabolome extraction procedure was performed as done at Glasgow Polyomics, University of Glasgow, Glasgow, UK (based on Creek et al 2011): i) 5 microL urine was extracted in 200 microL chloroform/methanol/water (1:3:1) at 4 C Celcius; ii) then vortexed for 5 minutes at 4 Celcius; iii) then centrifuged for 3 minutes (13,000 g) at 4 Celcius. The resulting supernatant was stored at -80 Celcius until analysis. A pooled aliquot of all 100 urine samples was prepared prior to the LC-MS/MS runs. Full details also on LC-MS/MS metabolomics data acquisition and analysis can be found in Van der Hooft et al. 2016, Metabolomics. Chromatography and Mass Spectrometry details: The samples were analysed using a Thermo Scientific Ultimate 3000 RSLCnano system (Thermo Scientific, CA, USA). The pHILIC separation was performed with a SeQuant ZIC-pHILIC column (150 x 4.6 mm, 5 micrometer) equipped with the corresponding pre-column (Merck KGaA, Darmstadt, Germany) - the column temperature was maintained at 25 C. A linear biphasic LC gradient was conducted from 80% B to 20% B over 15 min, followed by a 2 min wash with 5% B, and 8 min re-equilibration with 80% B, where solvent B is acetonitrile and solvent A is 20 mM ammonium carbonate in water. The flow rate was 300 microL/min, column temperature was held at 25 C, injection volume was 10 microL, and samples were maintained at 4 C in the autosampler. MS equipment and settings used: The LC system was coupled to a Thermo Scientific Q-Exactive Orbitrap mass spectrometer equipped with a HESI II interface (Thermo Scientific, Hemel Hempstead, UK). The set up was calibrated (Thermo calmix) in both ionization modes and tuned for the lower m/z range before analysis. Full scan (MS1) data was acquired in positive and negative switching mode in profile mode at 35,000 resolution (at m/z 200) using 1 microscan, an AGC target of 106 cts, a maximum injection time of 250 ms, with spray voltages +3.8 and -3.0 kV, capillary temperature 320 C, heater temperature of 150 Celsius, sheath gas flow rate 40 a.u., auxiliary gas flow rate 5 a.u., sweep gas flow rate 5 a.u, a full scan mass window of 70-1050 m/z, and using m/z 74.0964 (+) and m/z 112.98563 (-) as locking masses. Fragmentation data (LC-MS/MS) was obtained in positive ionization mode as described in (van der Hooft et al 2016). Briefly, a duty cycle consisted of one full scan (MS1) event and one Top5 (or Top10) MS/MS (MS2) fragmentation event, with full scan (MS1) resolution (at m/z 200) was set to 70,000, the AGC target set to 1 x 106, and the maximum injection time set to 120 ms. MS/MS (MS2) resolution (at m/z 200) was set to 17,500, the AGC target set to 2 x 105, MS/MS maximum injection time was set to 80 ms and the underfill ratio was set to 10 %, with a resulting intensity threshold of 2.5 x 105 cts.

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