A separation chromatographic method, already reported in the literature [38 (link)], was fully applied. In brief, each sample was injected three times. Analyses were performed at room temperature (22 ± 2 °C). A high-performance liquid chromatography (LC-20 AD VP; Shimadzu Corp., Kyoto, Japan), equipped with an ultraviolet (UV)–visible detector (Shimadzu Model SPD10 AV) set at λ 220 nm and Fluorescence detector (FLD) set at the excitation wavelength of 263 nm and at the emission wavelength of 305 nm was conducted. The analytical column was a reversed-phase LC column Kinetex phenyl-hexyl (100 Å, 150 × 4.6 mm, 5.0 μm particle size), equipped with a precolumn (4 × 3.0 mm) (Phenomenex, Torrance, CA, USA). All mobile phases were vacuum filtered through 0.45 μm nylon membranes (Millipore, Burlington, MA, USA). BPF, BPE, BPA, BPB, BPAF, BADGE and 4-NP were determined by FLD, while BPS, 2-CP, DCB, TCB, TCS and DEHP were Ultraviolet detected (UV). Data acquisition and integration were accomplished by Cromatoplus 2011 software. Methanol injections were made randomly to assess that no carryover occurred.
Kinetex phenyl hexyl
Manufactured by Phenomenex
Sourced in United States
The Kinetex Phenyl-Hexyl is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a variety of compounds. It features a phenyl-hexyl stationary phase that provides unique selectivity for a wide range of analytes.
Automatically generated - may contain errors
Lab products found in correlation
Nylon membrane, by Merck Group (1 mentions)
Lc 20advp, by Shimadzu (1 mentions)
Model spd10 av, by Shimadzu (1 mentions)
Lc ms grade methanol, by Carlo Erba (1 mentions)
Dionex ultimate 3000 rapid separation, by Thermo Fisher Scientific (1 mentions)
Qtof impact 2 mass spectrometer, by Bruker (1 mentions)
Microtof qiii mass spectrometer, by Bruker (1 mentions)
1290 infinity, by Phenomenex (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Ammonium formate, by Merck Group (1 mentions)
Ultrapure water, by Biosolve (1 mentions)
6 protocols using kinetex phenyl hexyl
1
Separation and Quantification of Endocrine Disruptors
2
Quantitative LC-MS Bioanalytical Assay
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Analytical procedures used 5 μl injections on a 2.6 μm Phenomenex Kinetex Phenyl Hexyl (50 × 4.6 mm) LC column heated to 35°C. Analytes were resolved at 0.5 mL/min using mobile phase A (10 mM ammonium formate in ultrapure 18.2 MΩ⋅cm water) and mobile phase B (0.1% formic acid in methanol). Analytes are resolved using a gradient starting at 95% aqueous (Mobile Phase A) and ramping to 0% aqueous over 4 min, and holding constant for 1 min. The gradient returned to initial conditions over 0.1 min and equilibrated for an additional 1.9 min. The total run time including column equilibration period between injections was 7.0 min. Specific mass spectrometer and analyte parameters are provided in Supplementary Tables S2 , S3 .
3
Targeted Metabolite Quantification by LC-MS/MS
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Chromatographic separation was achieved using a Phenomenex KineteX® Phenyl-Hexyl (50 × 4.6 mm, 2.6 µm) column coupled with a Phenomenex SecurityGuard Phenyl guard column (4.6 mm × 2.0 mm). The column temperature was controlled at 30 °C. Mobile phase A was 10 mM ammonium formate in water and phase B was 0.1% formic acid in methanol. The injection volume was 20 μL and the flow rate was 0.7 mL/min. The gradient condition was (time in minutes, % mobile phase B): (0,20), (0.7, 40), (4.5, 100), [6] (link), (6.1, 20) and (7.5, 20). Samples were analyzed by mass spectrometry in positive ion ESI mode. The source parameters were as follows: curtain gas of 30 psi, ion spray voltage of 2500 V, ion source temperature of 650 °C, medium collision gas, ion source gas 1 (GS1) of 50 psi and GS2 of 60 psi. MRM mode was utilized, and MRM parameters were the same as DMS-MS/MS method (Table 1 ). Target scan time was 0.5 s and all analytes were monitored within a + 0.5 min retention time window.
4
Extraction and Analysis of Plant Metabolome
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The surface metabolome was extracted in the laboratory by dipping each frond in 5 mL of LC-MS grade methanol (Carlo Erba, Peypin, France) during 5s according to [38 (link)] and subsequent sample preparation is detailed in SI. Analyses were performed on a UHPLC-ESI-HRMS system (Dionex Ultimate 3000 rapid Separation; ThermoFisher Scientific, Illkirch, France) with an analytical core-shell reversed phase column (150 × 2.1 mm, 1.7 μm, Kinetex Phenylhexyl; Phenomenex, Le Pecq, France) coupled with a QToF Impact II mass spectrometer (Bruker Daltonics, Bremen, Germany) in positive mode (details in SI).
5
Mass Spectrometric Analysis of C. nucifera Extracts
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C. nucifera extracts were analyzed following the methodology by Chang et al (2011)[11 (link)] with modifications using a Bruker Daltonics micrOTOF-QIII mass spectrometer (Bruker, Bremen, Germany) with atmospheric pressure chemical ionization (APCI) source in positive mode. The HPLC consisted of an Agilent Technologies (Waldbronn, Germany) 1290 Infinity with a binary pump, an autosampler, a diode-array detector on the 190–400 nm absorption region and a Kinetex Phenyl-Hexyl (2.1 x 50 mm, 1.7 μm, Phenomenex, Torrance, USA) column. The binary phase consisted of 0.1% formic acid in water (grade milli-q) (A) and methanol (grade LC-MS) (B) at the flow rate of 0.3 ml min-1. The elution profile was: 0–6 min, 20–65% B in A (linear gradient); 6–9 min, 65–100% B in A (linear gradient); 9–15 min, 100% B in A (isocratic; column wash); 15–18 min, 100–20% B in A (linear gradient; return to initial conditions). Positive ion mode APCI conditions were: corona current 5000 nA, capillary voltage 4000 V with the end plate offset at 500 V, drying gas (N2) temperature 300°C with the flow rate of 3.0 L min-1, nebulizer gas (N2) pressure 2.0 Bar, vaporizer temperature if 300°C and mass range for data acquisition 50–3,000 Da.
6
Quantitative Analysis of Novel Psychoactive Substances
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All chemicals and reagents used were of analytical grade. Methanol (≥ 99.9%) and acetonitrile (≥ 99.9%) were purchased from Merck (Darmstadt, Germany); ultrapure water was obtained from Biosolve Chimie (Dieuze, France). Formic acid (≥ 98%) and ammonium formate (97%) were purchased from Merck (Darmstadt, Germany). The analytical column Kinetex Phenyl-Hexyl (50 × 4.6 mm, 2.6 μm) was obtained from Phenomenex (Torrance, USA). Analytical standards consisting of NPS (Supplementary material-Table S1) were provided by the Italian National Institute of Health. Internal standard mix, consisting of deuterated compounds, was purchased from Chromsystems Instruments and Chemicals GmbH (Munich, Germany).
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