In the three colonies used for queen inhibition bioassays (G–I), we verified the presence of RNERO in mature NQs and other available castes. Individuals were extracted in clean glass vials using 30 µl of n-hexane (GC-MS grade, Merck) per individual (or per approx. 50 eggs) for 10 min at room temperature. Extracts were transferred into clean vials and stored at −20 °C. We used two-dimensional gas chromatography coupled with mass spectrometry (GC×GC-TOFMS, Pegasus 4D Leco, St. Joseph, MI, USA), equipped with a combination of non-polar ZB-5MS (30 m, internal diameter 0.25 mm, film thickness 0.25 μm, Phenomenex, Torrance, CA, USA) and medium polarity BPX-50 (1.3 m, internal diameter 0.1 mm, film thickness 0.1 μm, Restek, Bellefonte, PA, USA) columns. The temperature program for the primary column was 50 °C (1 min) to 320 °C (20 min) at 8 °C/min; the secondary column was set 10 °C higher. We identified RNERO based on Kovats retention index (1566), elution time comparison with synthetic standard and EI-MS fragmentation pattern39 (link). Relative RNERO abundance was compared by simultaneous visualization of all chromatograms from a given colony on an identical scale of detector intensities in Leco ChromaTOF software.
Zb 5ms
Manufactured by Phenomenex
Sourced in United States
The ZB-5MS is a gas chromatography (GC) column manufactured by Phenomenex. It is a fused silica capillary column coated with a 5% phenyl-95% dimethylpolysiloxane stationary phase. The ZB-5MS column is designed for the separation and analysis of a wide range of volatile and semi-volatile organic compounds.
Automatically generated - may contain errors
Lab products found in correlation
Gc 2010 plus, by Shimadzu (8 mentions)
Dxr raman microscope, by Thermo Fisher Scientific (4 mentions)
Trace dsq single quadrupole mass spectrometer, by Thermo Fisher Scientific (2 mentions)
6890n gas chromatograph, by Agilent Technologies (2 mentions)
Topspin software, by Bruker (2 mentions)
Quantum xls ultra ms, by Thermo Fisher Scientific (2 mentions)
Qp2010 ultra, by Shimadzu (2 mentions)
Gc ms system, by Agilent Technologies (2 mentions)
Gc ms grade, by Merck Group (1 mentions)
Qp2010 ultra mass spectrometer, by Shimadzu (1 mentions)
Mass hunter quantitative analysis b 08, by Agilent Technologies (1 mentions)
Trace 1310, by Thermo Fisher Scientific (1 mentions)
Trace gc ultra, by Thermo Fisher Scientific (1 mentions)
Gcms qp2010 se mass spectrometer, by Shimadzu (1 mentions)
Gc 2010 plus instrument, by Shimadzu (1 mentions)
Trace 1310 gc, by Thermo Fisher Scientific (1 mentions)
Scion 456 gc, by Techcomp Instruments (1 mentions)
Supelco 37 component fame mix, by Merck Group (1 mentions)
Trace dsq, by Thermo Fisher Scientific (1 mentions)
As 3000, by Thermo Fisher Scientific (1 mentions)
33 protocols using zb 5ms
1
Queen Inhibition Bioassay for RNERO
2
GC-MS Analysis of Essential Oils
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GC–MS was performed with a Shimadzu GC-2010 Plus coupled to a Shimadzu QP2010 Ultra mass spectrometer (Japan). A fused-silica capillary column ZB-5 MS (30 m, 0.25 mm i.d.) with a film thickness of 0.25 mm (Phenomenex, Torrance, CA, USA) was used. GC-MS conditions applied for analysis of the essential oil and obtained fractions followed Sieniawska et al. [12 (link)]. The retention indices were determined in relation to a homologous series of n-alkanes (C8–C24) under the same operating conditions. Constituents of studied essential oil were identified by comparison of their mass spectra and retention indices with computer-supported spectral libraries (Mass Finder 2.1 and NIST database) and with authentic standard of β-elemene.
3
Fatty Acid Dimethyldisulfide Adduct Analysis
4
GC-MS Analysis of Volatile Metabolites
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A TRACE DSQ single quadrupole mass spectrometer from Thermo Fisher was used to perform the GC‐MS analyses, based on the way developed by Verslues (2017 ) with faint modifications. The GC conditions used were as follows: column: ZB‐5MS (Phenomenex), 0.25 µm film thickness, 30 mm × 0.25 mm; carrier gas: helium; split flow: 10 ml/min; linear velocity: injector temperature: 230°C; constant flow rate at 1.3 ml/min; and column temperature program: start temperature (40°C) held for 1 min, after which, increasing to 310°C at 5°C/min. There are MS conditions: ionization: detection: positive ion; electron impact (70 eV); full scan analyses: 10–600 m/z at the rate of two scans per second. Volatile metabolites were eluted by using the solvent front in this method, so GC separation of these analyses began with an initial start temperature at 40°C which was maintained for 2 min. The temperature was increased to 80°C at 10°C/min. The temperature was then kept constant at 80°C for 3 min before increasing to 230°C at a rate of 30°C/min.
5
GC-MS Analysis of Ambergris Extracts
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Extracts were analysed using a Thermo Fisher Trace 1310 gas chromatograph coupled to a Thermo Fisher Quantum XLS Ultra mass spectrometer (ThermoFisher Scientific, Waltham, Massachusetts, USA). A ZB-5ms (Phenomenex, Torrance, USA), 30 m fused silica capillary column, 250 µm i.d. with a film thickness of 0.25 µm was used with helium (BIP® ECD) as carrier gas at a constant flow of 1.5 mL min−1. The ambergris extract was injected in split mode 20:1; all other samples were injected splitless. The injection volume was 1 µL. The injection port was raised from 80 °C to 300 °C at 14.5 °C s−1. The column temperature was kept at 80 °C for 1 min, then increased to 310 °C at 5 °C min−1 where it was kept for 20 min. Solvent delay for filament activation of the ion source was set at 20 min and electron ionization voltage was set at 70 eV. The scanning mass range was m/z 50–600. Steroids and ambrein were identified according to their TMS-mass spectra published in the literature3 (link),19 ,29 (link), the NIST 05 Mass Spectral Search Program and by comparing mass spectra and elution order with steroid analytical standards (from Sigma Aldrich (Germany) and Chiron (Norway)) for coprostanol, epicholestanol, epicoprostanol, 5α-cholestanol and cholesterol.
6
GC-MS Analysis of Essential Oil Components
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The chemical components of the analyzed essential oil were separated by the gas chromatography and identified by the mass spectrometry using Thermo Scientific GC–MS system (DSQ II GC/MS with Trace GC Ultra, Palo Alto, USA). Samples were analyzed on the capillary column ZB-5 MS 30 m × 0.25 mm, film thickness 0.2 μm, (Phenomenex, Torrance, USA). GC–MS settings were as follows: the initial oven temperature was held at 60 °C for 1 min and ramped at 4 °C/min rising to 250 °C; helium was used as carrier gas at a flow rate of 1 mL/min; the sample (0.1 μL) was injected manually at the split/splitless injector temperature of 260 °C, with a split ratio 1:50 and transfer line temperature was set to 270 °C for GC–MS analyses. Mass spectra were obtained at 70 eV (EI), the scan range was 45–350 m/z and Xcalibur version 2.0.7. was used for result processing and quantification. The components of the essential oil were identified by obtained GC–MS spectra and retention indices (RI) relative to C8-C20 n-alkanes. AMDIS computer program version 2.62 was used for GC–MS data processing using NIST library version 2.0. Spectra and obtained retention indices were compared with data already available in the literature42 .
7
GC-EIMS Analysis of Chemical Compounds
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Gas chromatography-Electro ionization Mass Spectrometry (GC–EIMS) analysis was performed on a fused silica column (ZB-5-MS, 5% phenyl methyl polysiloxane, 30 m, 0.25 mm i.d., 0.25 µm film thickness; Phenomenex, Torrance, CA, USA) in a GC 2010 chromatograph coupled with GCMS-QP2010 SE mass spectrometer (Shimadzu, Kyoto, Japan) equipped with a quadrupole analyzer. Helium was used as a carrier gas at the flow rate of 1 mL/min. The programmed temperature was set as follows: oven temperature was raised from 100 °C to 325 °C using a ramp of 20 °C/min. The final temperature was maintained for 5 min (end of the analysis). One µL of the sample was injected into the column with a split ratio of 1/10. The MS detector was set as follows: electron impact mode (70 eV) with the ion source temperature set at 200°C, analyzed mass range m/z 40–700. Spectrum was acquired from 3 min (solvent delay) to 16.25 min (end of the run). The identification of the chemical structures was performed by comparison with a library of mass spectra (NIST MS Search 2.0).
8
Comprehensive Spectroscopic Analysis Protocol
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1H‐NMR spectra were recorded on Bruker spectrometers in C2HCl3 at 500 MHz or 300 MHz for 1H and at 126 MHz or 100 MHz for 13C. 2H‐NMR spectra were recorded in C1HCl3 at 77 MHz or 61 MHz. All spectra were analysed by Bruker TopSpin software. Raman spectra were measured on crystalline samples or on the neat liquid supported on glass microscope slides using a ThermoScientific DXR Raman Microscope at laser frequencies of either 532 nm or 780 nm. Infrared spectra were recorded for crystalline or neat liquid samples on a sapphire anvil using an Agilent Resolutions Pro spectrometer. GC‐MS analysis was performed using 1‐μl injections of solutions at approximately 1 mg/ml in dichloromethane using an Agilent 6890N gas chromatograph coupled to a 5973 mass selective detector. The column was a Phenomenex ZB‐5MS, 30 m × 0.25 mm, used with a temperature profile: 50°C (held for 3 min), ramp at 10°C per minute to 250°C and held for 2 min. The carrier gas was helium at 1 ml/min. The molecular formulae for the labelled compounds were determined at the National Mass Spectroscopy Facility, University of Swansea, using an LTQ Orbitrap XL operating in APCI or NSI modes.
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1H‐NMR spectra were recorded on Bruker spectrometers in C2HCl3 at 500 MHz or 300 MHz for 1H and at 126 MHz or 100 MHz for 13C. 2H‐NMR spectra were recorded in C1HCl3 at 77 MHz or 61 MHz. All spectra were analysed by Bruker TopSpin software. Raman spectra were measured on crystalline samples or on the neat liquid supported on glass microscope slides using a ThermoScientific DXR Raman Microscope at laser frequencies of either 532 nm or 780 nm. Infrared spectra were recorded for crystalline or neat liquid samples on a sapphire anvil using an Agilent Resolutions Pro spectrometer. GC‐MS analysis was performed using 1‐μl injections of solutions at approximately 1 mg/ml in dichloromethane using an Agilent 6890N gas chromatograph coupled to a 5973 mass selective detector. The column was a Phenomenex ZB‐5MS, 30 m × 0.25 mm, used with a temperature profile: 50°C (held for 3 min), ramp at 10°C per minute to 250°C and held for 2 min. The carrier gas was helium at 1 ml/min. The molecular formulae for the labelled compounds were determined at the National Mass Spectroscopy Facility, University of Swansea, using an LTQ Orbitrap XL operating in APCI or NSI modes.
9
GC-MS Analysis of Essential Oils
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A Shimadzu GC-2010 Plus instrument coupled to a Shimadzu QP2010 Ultra mass spectrometer (Shim-pol, Warsaw, Poland) was used for GC-MS analyses. The chromatograph was equipped with a fused-silica capillary column ZB-5 MS (30 m, 0.25 mm i.d.) with a film thickness of 0.25 mm (Phenomenex, Torrance, CA, USA). The oven temperature program was started at 50 °C, held for 3 min, then increased at the rate of 8–250 °C/min, and held for a further 2 min. The MS was operated in EI mode; the scan range was 40–500 amu, the ionization energy was 70 eV, and the scan rate was 0.20 s per scan. The injector (250 °C), interface (250 °C), and ion source (220 °C) temperatures were set. Split injection was performed with a split ratio of 1:20. Helium was the carrier gas at a 1.0 mL/min flow rate. Each of the 9 EOs samples were prepared by diluting 2 µL of EO in 1 mL of hexane. An internal standard was added to each sample. Three parallel measurements were done. The relative percentages of each component present in the EOs were calculated. The retention indices were determined in relation to a homologous series of n-alkanes (C8–C24) under the same operating conditions. The compounds were identified with computer-assisted spectral libraries (MassFinder 2.1 Hamburg, Germany; NIST 2011, Gaithersburg, MD, USA).
10
GC-MS Analysis of Organic Compounds
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Compounds were analysed with a Thermo Fisher Trace 1310 GC coupled to a Thermo Fisher Quantum XLS Ultra MS. Fractions were injected with an autosampler (Thermo TriPlus RSH) onto a fused-silica capillary column (Phenomenex Zebron ZB-5MS, 30 m, 0.1 μm film thickness, inner diameter 0,25 mm). The carrier gas was He at a flow rate of 1.5 mL/min. The GC oven temperature was ramped from 80°C (1 min) to 310°C at 5°C/min and held for 20 min. Electron ionization mass spectra were recorded in full scan mode at an electron energy of 70 eV within a mass range of m/z 50 to 600.
Compounds were identified by their retention times and comparison with published mass spectra. FAME were characterized by comparison of mass spectra and retention times with an external standard mixture (Supelco 37 FAME standard). Compounds were quantified by comparison with internal standards of known concentration.
Compounds were identified by their retention times and comparison with published mass spectra. FAME were characterized by comparison of mass spectra and retention times with an external standard mixture (Supelco 37 FAME standard). Compounds were quantified by comparison with internal standards of known concentration.
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