All samples were analyzed using an Agilent Technologies 7890A GC system coupled to an Agilent Technologies 5975 inert XL EI/CI Mass Selective Detector. The samples were injected in splitless mode with an injection volume of 1 µL. The separation of the analytes was performed on a 30-m Zebron ZB5 capillary column (Phenomenex, 0.25-mm inner diameter × 0.25-µm film thickness). Helium was used as the carrier gas at a constant flow of 1.1 mL/min resulting in an average velocity of 38.2 cm/s. The initial temperature was 50 °C, which was held for 1 min and hereafter increased by 25 °C/min until 100 °C, followed by an increase to 315 °C with 12 °C/min. The temperature was kept at 315 °C for 7 min, which resulted in a total analysis time of 28 min. Electron ionization (EI) was performed using an electron energy of 70 eV. The temperature of the EI source was 230 °C and the quadrupole 150 °C. The mass spectrometer was operated in SIM mode for 21 selected mass-to-charge (m/z) ratios, ranging from 83 to 288 m/z. The data acquisition was divided into 12 SIM groups with a dwell time of 20 ms of each m/z (table S7.1 ).
Zb 5 capillary column
Manufactured by Phenomenex
Sourced in United States, Austria
The ZB-5 capillary column is a chromatographic column designed for gas chromatography applications. It features a 5% phenyl-95% dimethylpolysiloxane stationary phase, which provides a balance of polarity and thermal stability. The ZB-5 column is suitable for the separation and analysis of a wide range of organic compounds.
Automatically generated - may contain errors
Lab products found in correlation
Gc 2010, by Shimadzu (7 mentions)
Trace gc 2000, by Thermo Fisher Scientific (2 mentions)
7890a gc system, by Agilent Technologies (1 mentions)
5973n mass selective detector, by Agilent Technologies (1 mentions)
6890 series gas, by Agilent Technologies (1 mentions)
Msd chemstation software, by Agilent Technologies (1 mentions)
Clarus 580 gas chromatograph, by PerkinElmer (1 mentions)
Jms t100cs, by JEOL (1 mentions)
V 770, by Jasco (1 mentions)
Zb waxplus capillary column, by Phenomenex (1 mentions)
Jnm la400, by JEOL (1 mentions)
Gcms qp2010, by Shimadzu (1 mentions)
Ft ir 4100, by Jasco (1 mentions)
Gc 2014, by Shimadzu (1 mentions)
Flame ionization detector, by Shimadzu (1 mentions)
Qp5050a, by Shimadzu (1 mentions)
Model 5975 mass spectrometer, by Agilent Technologies (1 mentions)
Turbomass, by PerkinElmer (1 mentions)
Vacuum manifold, by Merck Group (1 mentions)
20 protocols using zb 5 capillary column
1
GC-MS Quantification of Analytes
2
GC-MS Analysis of Crude Extracts
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Crude extracts were analyzed by gas chromatography—mass spectrometry, using a Trace GC2000 (Thermo, Austria), equipped with a ZB-5 capillary column (30 m × 0.25 mm i.d., × 0.25 μm film thickness (Phenomenex, Germany) at a helium flow of 1.2 ml min−1. The temperature of the GC oven was raised from 50°C (1 min) to 300°C at 10°C min−1, then held for 5 min at 300°C. The GC was coupled to a DSQ I MS (ion source at 200°C; transfer line at 310°C). All mass spectra were recorded in EI mode. Components adsorbed on SPME-fibers were thermally desorbed in the hot GC injector at 240°C, and analyzed as described above. Retention indices (RI) were calculated using an alkane standard mix according to the formula RIx = 100n0 + (100tx−100tn0)/(tn1−tn0), with x: target compound; tx: retention time of target compound; n0: number of carbon atoms in the alkane directly eluting before x; tn0: retention time of alkane directly eluting before x; tn1: retention time of alkane directly eluting after × (Van den Dool and Kratz, 1963 (link)).
3
GC-MS Analysis of Organic Compounds
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GC-MS analyses were carried out on an Agilent 6890 Series gas chromatograph equipped with a 5973 N mass selective detector (USA). A programmed temperature vaporization injector operating in the split mode (1:100) at 250 °C and a ZB-5 capillary column (30 m × 0.25 mm i.d. 0.25 μm film thickness; 5% diphenyl, 95% dimethylpolysiloxane; Phenomenex, USA) were used. The increase in the oven temperature was programmed from 80 °C (held for 1 min) to 300 °C (held for 10 min), at a rate of 20 °C/min. Helium was used as carrier gas at constant pressure (9.8 psi) and the injection volume was 1 μL. The transfer line, ion source and quadrupole temperatures were maintained at 280, 230 and 150 °C, respectively, and a solvent delay of 4 min was selected. Electron ionization was performed at 70 eV and the mass spectrometer was operated in the full scan mode in the range 35–550 Da. All results were compared with Wiley’s library reference spectral bank (G1035B; RevD.02.00). Data recording and instrument control were performed by the MSD ChemStation software (G1701 CA; ver.C.00.00; Agilent Technologies, Little Falls, DE, USA.
4
Monosaccharide Analysis of LOS Samples
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The monosaccharides were analyzed as acetylated methyl glycosides by GC-MS [14 (link)]. Briefly, an aliquot of LOS samples (0.5 mg) from the cells and EMVs were subjected to a methanolysis reaction with HCl/CH3OH (1.25 M, 1 mL) at 80 °C for 16 h. The mixture was extracted three times with hexane. The hexane phases containing the fatty acids methyl esters were directly analyzed by GC-MS. The methanol layers, containing the methyl glycosides were dried and then acetylated with acetic anhydride in pyridine at 100 °C for 30 min.
All the samples were analyzed on an Agilent Technologies gas chromatograph 6850A equipped with a mass selective detector 5973N and a Zebron ZB-5 capillary column (Phenomenex, Bologna, Italy 30 m × 0.25 mm i.d., flow rate 1 mL/min, He as carrier gas). Acetylated methyl glycosides were analyzed using the following temperature program: 140 °C for 3 min, then 140→240 °C at 3 °C/min. The temperature program for methyl esters of fatty acids was the following: 140 °C for 3 min, then 140→280 °C at 10 °C/min, and finally 280 °C for 20 min.
All the samples were analyzed on an Agilent Technologies gas chromatograph 6850A equipped with a mass selective detector 5973N and a Zebron ZB-5 capillary column (Phenomenex, Bologna, Italy 30 m × 0.25 mm i.d., flow rate 1 mL/min, He as carrier gas). Acetylated methyl glycosides were analyzed using the following temperature program: 140 °C for 3 min, then 140→240 °C at 3 °C/min. The temperature program for methyl esters of fatty acids was the following: 140 °C for 3 min, then 140→280 °C at 10 °C/min, and finally 280 °C for 20 min.
5
GC-FID Cholesterol Quantification Protocol
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Separation was carried out using a Clarus 580 Gas Chromatograph (Perkin Elmer, Waltham, MA, USA) fitted with a flame ionization detector (GC-FID) set at 260 °C according to [26 (link)]. These authors previously showed the method to have a limit of quantitation of 0.1 µg/mL (equivalent to 0.4 mg cholesterol per g meat); a limit of detection was not reported. The separation and quantification of cholesterol was carried out employing a ZB-5 capillary column (Phenomenex, Torrance, CA, USA) with a film thickness of 0.25 µm and a length of 30 m × 0.25 mm. The injection volume was 0.5 µL and the inlet temperature was set to 200 °C. Hydrogen was flushed at a constant flow of 2.0 mL/min, and the split ratio was set at 5:1. The oven temperature started at 180 °C with an initial temperature ramp of 8.0 °C/min to 260 °C followed be a second temperature ramp of 2 °C/min to 280 °C which was held for 10 min.
The content of cholesterol was calculated using following equation (Equation (1)) according to Grasso et al. [26 (link)].
where, IS Purity is the purity of the internal standard as given on the certificate of analysis, RRF is the relative response factor for cholesterol (namely 1.001), and 20 is the dilution factor.
The content of cholesterol was calculated using following equation (Equation (1)) according to Grasso et al. [26 (link)].
where, IS Purity is the purity of the internal standard as given on the certificate of analysis, RRF is the relative response factor for cholesterol (namely 1.001), and 20 is the dilution factor.
6
GC-MS Analysis of Crude Extracts
7
Structural Analysis of LPS Bv Monosaccharides
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LPSBv monosaccharide content was established
by analysis of the acetylated O-methyl glycoside
derivatives obtained by treatment with HCl/MeOH (1.25 M, 85 °C,
24 h) followed by an acetylation step with acetic anhydride in pyridine
(85 °C, 30 min). The absolute configuration of each sugar unit
was defined through the evaluation of the O-octylglycoside
derivatives as previously described.22 (link) The
sugar linkage pattern was determined by the Ciucanu method:23 (link),24 (link) briefly, an aliquot of sample was suspended in DMSO to which NaOH
in powder was added and then methylated with CH3I, hydrolyzed
with trifluoroacetic acid (4 M, 100 °C, 4 h), carbonyl reduced
with NaBD4, and acetylated with pyridine and acetic anhydride.
The total fatty acid content was established on intact LPS by treating
with HCl (4 M, 100 °C, 4 h) followed by NaOH (5 M, 100 °C,
30 min). The pH was adjusted to reach slight acidity. After extraction
in chloroform, fatty acids were then methylated with diazomethane.25 (link) All chemical analyses were performed by means
of a gas–liquid chromatography (GLC-MS) Agilent Technologies
6850A instrument equipped with a mass selective detector 5973N and
a Zebron ZB-5 capillary column (Phenomenex, 30 m × 0.25 mm i.d.,
flow rate 1 mL/min, He used as carrier gas) and using the following
temperature program: 140 °C/240 °C at 3 °C/min.
by analysis of the acetylated O-methyl glycoside
derivatives obtained by treatment with HCl/MeOH (1.25 M, 85 °C,
24 h) followed by an acetylation step with acetic anhydride in pyridine
(85 °C, 30 min). The absolute configuration of each sugar unit
was defined through the evaluation of the O-octylglycoside
derivatives as previously described.22 (link) The
sugar linkage pattern was determined by the Ciucanu method:23 (link),24 (link) briefly, an aliquot of sample was suspended in DMSO to which NaOH
in powder was added and then methylated with CH3I, hydrolyzed
with trifluoroacetic acid (4 M, 100 °C, 4 h), carbonyl reduced
with NaBD4, and acetylated with pyridine and acetic anhydride.
The total fatty acid content was established on intact LPS by treating
with HCl (4 M, 100 °C, 4 h) followed by NaOH (5 M, 100 °C,
30 min). The pH was adjusted to reach slight acidity. After extraction
in chloroform, fatty acids were then methylated with diazomethane.25 (link) All chemical analyses were performed by means
of a gas–liquid chromatography (GLC-MS) Agilent Technologies
6850A instrument equipped with a mass selective detector 5973N and
a Zebron ZB-5 capillary column (Phenomenex, 30 m × 0.25 mm i.d.,
flow rate 1 mL/min, He used as carrier gas) and using the following
temperature program: 140 °C/240 °C at 3 °C/min.
8
GC-FID Analysis of B. ogadensis Oleogum Resin
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The B. ogadensis oleogum resin essential oils were analyzed by GC-FID using a Shimadzu GC 2010 (Shimadzu Scientific Instruments, Columbia, MD, USA) equipped with flame ionization detector, a split/splitless injector, and Shimadzu autosampler AOC-20i (Shimadzu Scientific Instruments, Columbia, MD, USA), with a ZB-5 capillary column (Phenomenex, Torrance, CA, USA) as previously described [27 (link)].
9
Extraction and Characterization of Cotton Gossypium Tomentosum Essential Oil
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Essential oil was extracted from pesticide-free CGT, which was comprised mainly of the calyx (bracts). Pesticide-free CGT was kindly supplied by the Australian Cotton Research Institute, Narrabri, NSW, Australia. Milled samples were extracted by means of hydro-distillation, as outlined in Egbuta et al. [50 (link)], and extracted oil was collected into a clean 22 mL vial. Extracted CGT oil was stored in the refrigerator at 4 °C for further use. In order to determine the chemical composition of the extracted oil, 50 µL (45.6 mg) CGT oil was diluted in 950 µL (623.9 mg) hexane and analyzed using an Agilent 6890A GC instrument (Wilmington, DE, USA) equipped with a ZB-5 capillary column (Phenomenex) with the dimensions 30 m length × 0.53 mm internal diameter (I.D) × 1.50 µm film thickness. Conditions for the semi-quantitative analysis were the same as the conditions applied by Egbuta et al. [50 (link)].
10
Characterization of Vanadium Compounds
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The cold-spray ionization mass spectra were recorded on a JEOL JMS-T100CS. NMR spectra were recorded with a JEOL JNM-LA400. 51V NMR spectra were measured at 105.15 MHz. Chemical shifts were referenced to VOCl3 (δ = 0 ppm) for 51V NMR. IR spectra were measured on a Jasco FT/IR-4100 using a nujol method. Elemental analyses of C, H, and N were performed by the Research Institute for Instrumental Analysis at Kanazawa University. GC analysis was performed on a Shimadzu GC-2014 with a flame ionization detector (FID) equipped with a ZB-WAXplus capillary column (phenomenex, internal diameter = 0.25 mm, length = 30 m), NEUTRABOND capillary column (GL science, internal diameter = 0.25 mm, length = 30 m), and ZB-5 capillary column (phenomenex, internal diameter = 0.25 mm, length = 30 m). Naphthalene was used as the internal standard. GC-MS spectra were measured on a Shimadzu GCMS-QP2010. UV/vis spectra were measured on a Jasco V-770.
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