A millipede or 10 μl of blood were transferred into a glass vial and immersed in n-hexane (5 ml) for 3 min at room temperature. The n-hexane extract (4 μl portion, each) was analyzed using a gas chromatography-mass spectrometer (GC/MS) (7890A GC System coupled with a 5975C inert XL EI/CI MSD with a Triple-Axis Detector operated at 70 eV; Agilent Technologies, Santa Clara, CA, USA) equipped with an HP-5 ms capillary column (0.25 mm i.d. × 30 m, 0.25 μm film thickness; Agilent Technologies) according to the previous publication13 (link).
5975c inert xl ei ci msd
Manufactured by Agilent Technologies
Sourced in United States
The 5975C inert XL EI/CI MSD is a gas chromatography-mass spectrometry (GC-MS) system designed for high-performance analysis of a wide range of chemical compounds. It features an inert ion source and collision cell, providing enhanced sensitivity and reduced matrix effects for reliable quantitation and identification. The system is capable of both electron ionization (EI) and chemical ionization (CI) techniques, offering versatility in analytical applications.
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Lab products found in correlation
7890a gc system, by Agilent Technologies (6 mentions)
Hp 5ms capillary column, by Agilent Technologies (2 mentions)
Triple axis detector, by Agilent Technologies (2 mentions)
Gentamycin, by Thermo Fisher Scientific (1 mentions)
Db 17ms column, by Agilent Technologies (1 mentions)
Reliaprep blood gdna miniprep system, by Promega (1 mentions)
Poraplot q 25 0.32 column, by Agilent Technologies (1 mentions)
7820a gas chromatograph, by Agilent Technologies (1 mentions)
Inova 500 mhz, by Agilent Technologies (1 mentions)
8453 uv vis spectrophotometer, by Agilent Technologies (1 mentions)
Gc sampler 80, by Agilent Technologies (1 mentions)
10 protocols using 5975c inert xl ei ci msd
1
Millipede and Blood Chemical Profiling
2
Sebocyte 3D Culture and Fatty Acid Analysis
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Human SEBO662AR sebocytes were seeded into culture inserts and grown in culture medium (K‐SFM supplemented with 25 µg mL−1 of gentamycin, Thermo Fisher Scientific) for 48 hours at 37°C with 5% CO2. After this immersion culture, the cultures were placed at the air–liquid interface (D0) and the medium was replaced by culture medium containing or not containing (for the control) the compounds being tested or a non‐androgenic seborrhoeic reference and the cultures were incubated for 7 days with renewal of the culture medium and treatments (Vetiver extract at 1% (v/v) and reference) every 2 to 3 days. All the experimental conditions were performed in triplicate. The 3D tissues were cut, put in microtubes and dry frozen at −80°C. The determination of the content of Free Fatty Acids is performed using a GC (7890A; Agilent, Santa Clara, California, USA) coupled with a MS system (5975C Inert XL EI/CI MSD, Agilent). The extraction of fatty acids was performed using a liquid/liquid extraction (proportions are the propriety of BioAlternatives). Samples were then methylated (FAME) before analysis.
3
GC-MS Analysis of Microalgal Fatty Acids
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Fatty acid composition of all microalgal extracts was analyzed by GC–MS by using an Agilent 7890A connected to an Agilent 5975C Inert XL EI/CI MSD (Palo Alto, CA, USA). Prior to analysis, fatty acid methyl esters (FAMEs) were freshly prepared by base-catalyzed methanolysis of the glycerides (KOH in methanol). FAMEs were separated by using an HP-5ms ultra-inert column (30 m × 250 µm × 0.25 µm) (Palo Alto, CA, USA). A total of 1 µL of the sample was injected in spitless mode and an injector temperature of 280 °C. The initial oven temperature was set at 150 °C for 1 min and the temperature was gradually raised to 220 °C at 3 °C/min with a final increase to 300 °C for 3 min. Helium was used as carrier gas at a constant column flow rate of 2.52 mL/min. The GC to MS interface temperature was fixed at 280 °C and an electron ionization system was set on the MS in scan mode. The mass range evaluated was 50–600 m/z, where the MS quad and source temperatures were maintained at 150 °C and 230 °C respectively. Fatty acids were identified by comparing their retention times and mass spectrum profiles with known standards (FAME mix supelco) and the NIST mass spectral library (Version 2.2).
4
Deuterium Labeling for DNA Turnover
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Genomic DNA was isolated from sorted cell samples (typically consisting of >250,000 cells) using the ReliaPrep Blood gDNA Miniprep system (Promega) according to manufacturer’s instructions. 2H incorporation in DNA was measured by gas chromatography and mass spectrometry (GC/MS) as previously described (16 (link)) with minor modifications. Briefly, after enzymatic hydrolysis of the DNA, purine deoxyribonucleotides were derivatized to perfluorotriacetyl (PFTA) and injected into the gas chromatograph (DB-17MS column; 7890A GC System, Agilent Technologies). The mass of the derivate was measured by negative chemical ionization mass spectrometry (5975C inert XL EI/CI MSD; Agilent Technologies) at m/z 435 (M0) and 436 (M1). Standards of known isotopic enrichments were used to control for varying sample concentrations, as reported previously (17 (link)). Plasma was obtained by centrifugation of whole blood samples and deuterium enrichment was measured on the same GC/MS system (using a PoraPLOT Q 25 × 0.32 column, Varian) as described by Westera et al. (18 (link)).
5
Deuterium-Labeled Compound Feeding in C. hualienensis
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To elucidate the (R)‐MAN biosynthetic pathway of C. hualienensis, deuterium‐labelled compounds were administered to millipedes by ‘force feeding’. Fifth stadium of C. hualienensis were each separately placed in a microvolume insert containing 4–5 μL of 1000 ppm aq. D5‐(E/Z)‐PAOx or D5‐PAN with their heads down, as shown in Fig. 1 A. After force feeding at room temperature for 16 h, the defensive compounds were extracted after the addition of n‐hexane (80–100 μL) for 3 min, and 4 μL of the extract were analysed using a GC–MS (7890A GC System coupled with a 5975C inert XL EI/CI MSD with a Triple‐Axis Detector operated at 70 eV; Agilent Technologies, Santa Clara, CA, USA) system equipped with an HP‐5 ms capillary column (0.25 mm i.d. × 30 m, 0.25 μm film thickness; Agilent Technologies) according to a previous report 55 .
6
GC-MS Analysis of Organic Compounds
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The GC-MS analysis was performed using Agilent Technologies 7890A capillary gas chromatograph, directly coupled to a mass spectrometer system (Model: 5975C inert XL EI/CI MSD with triple axis detector). A fused silica capillary column of 5% phenyl, 95% dimethyl-poly-silloxane (HP-5MSI; length: 90 m, diameter: 0.250 mm and film: 0.25 μm) was used. The GC parameter was set as follows: the inlet temperature was set at 250°C and oven temperature was programmed as 90°C for 0 min, then 3°C/min to 200°C for 2 min and then 15°C/min to 280°C for 2 min. Total run time was 46 min and column flow rate was 1.1 mL/min Helium gas. The auxiliary (GC to MS interface) temperature was set to 280°C. The MS parameter was set as the MS was in scan mode. The ionization mode was EI (electron ionization) type. The mass range was set in the range of 50–550 m/z. MS quad temperature and source temperature was set at 150°C and 230°C respectively. Each component were searched and identified by using “NIST-MS Library 2009.” Peak area of the total ionic chromatogram (TIC) was used to determine the relative percentage amounts of separated compounds and calculation were done automatically.
7
Volatile Organic Compound Profiling
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This was conducted at 3 and 5 months incubation. Microcosms were opened for 1 h in a sealed bell jar with a StableFlex™ 2 cm SPME fiber used to capture volatiles. The samples were immediately loaded by direct injection for GCMS analysis on an Agilent 7890A GC system with 5975C inert XLEI/CI MSD with triple axis detector and the 7683 Injector (Agilent, USA). The protocol for volatile samples was: from a starting point of 80 °C, temperature was ramped to 280 °C, at a rate of 5 °C increase per minute for a run time of 40 min with a final ramp to 300 °C at a rate of 20 °C increase per minute with a hold time of 4 min. The total run time was 45 min.
8
Spectroscopic Characterization and X-Ray Crystallography
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Infrared spectra were collected on a Nicolet Avatar 360 FTIR spectrometer. UV-Visible spectroscopy was performed in a 1 cm quartz cell using an Agilent Technology 8453 UV-Vis spectrophotometer equipped with a diode-array detector. NMR spectra were recorded a Bruker DPX 400 MHz and Varian Inova 500 MHz spectrometers in CDCl 3 or CD 3 CN solvent using standard conditions, and were referenced to the residual proton signal of the solvent. Elemental analysis was performed on a 4.1 Vario EL 3 elemental analyzer from Elementar. The ESI-MS experiments were performed with a Bruker Esquire 6000 LC/MS chromatograph, using acetonitrile as a mobile phase. The product analyses after catalysis experiments were carried out on an Agilent Technology 7820A gas chromatograph equipped with a 16-sample automatic liquid sampler, flame ionization detector and EzChrom Elite Compact software. GC-MS analyses were performed on an Agilent Technology 7890A GC system equipped with a 5975C inert XL EI/CI MSD with triple-axis detector. The products were identified by comparison of their GC retention times and, in the case of GC/MS, with those of authentic compounds. Colourless crystals of 1 Pz were grown from slow diffusion of ethyl ether in a CH 2 Cl 2 solution of the compound, and used for low temperature (100(2) K) X-ray structure determination.
9
Metabolite Extraction from Resin-Amended Microcosms
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This was conducted at 2, 3 and 5 months incubation. 1 g of autoclaved Amberlite XAD16N resin was added to the microcosm and incubated overnight. The resin was gently scraped off each microcosm using a sterile scalpel into separate Falcon 15 ml conical centrifuge tubes. 10 ml 50:50 ethyl acetate: methanol was added to each tube and placed on a rotating platform at 120 rpm for 1 h to extract metabolites before being dried completely in a centrifugal evaporator using the –OH setting at room temperature. The dried samples were stored in a − 80 °C freezer until ready for analysis. 90 μl ethyl acetate was used to reconstitute the sample in glass Chromacol vials immediately prior to GCMS analysis on an Agilent 7890A GC system with 5975C inert XLEI/CI MSD with triple axis detector (Agilent, USA). The reconstituted samples were spiked with 10 μl of 0.001% dodecane in ethyl acetate to act as an internal control for sensitivity and detection.
The GCMS protocol was: from a starting point of 80 °C for 2 min, temperature was ramped to 280 °C, at a rate of 10 °C increase per minute with a hold time of 4 min for a run time of 26 min with a final ramp to 320 °C at a rate of 20 °C increase per minute with a hold time of 1 min. The total run time was 29 min.
The GCMS protocol was: from a starting point of 80 °C for 2 min, temperature was ramped to 280 °C, at a rate of 10 °C increase per minute with a hold time of 4 min for a run time of 26 min with a final ramp to 320 °C at a rate of 20 °C increase per minute with a hold time of 1 min. The total run time was 29 min.
10
Aroma Profiling of Herbal Compresses
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Fresh herbal compress samples (0.5 g) were quickly cut in small pieces and put into 20 mL GC-HS vial for aroma volatile injection with the HS technique. GS analyses were performed with Agilent GC 7890A GS system equipped with a fused silica capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness: Mega-5MS). Injector and transfer line temperatures were 200 and 270°C, respectively. The initial temperature was 60°C for 1 min, raised to 270°C at the rate 4°C/min, and maintained at this temperature for 5 min. Helium (99.999%) was the carrier gas with a flow rate of 1 mL/min at constant flow. The number of samples in each analysis was equal to three (n = 3). Each fresh herbal compress sample which included C. dubia, C. citratus, T. indica, C. longa, Z. montanum, and C. hystrix was placed in an incubation at 80°C for 20 min, agitated at 500 rpm, and injected using the splitless mode at the split ratio of 1:10 and an autosampler (Agilent GC Sampler 80). For GC-MS detection, the GC system was interfaced with an Agilent 5975C inert XL EI/CI MSD; Triple-Axis detector unit operated under the EI mode (70 eV) with an ion source where the temperature was maintained at 230°C and a quadrupole analyzer (150°C) at a mass range of 40.0-900.0 amu. The dried herbal compress samples were subject to the same method as described.
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