Total IgE (BioLegend, San Diego, CA, United States), HDM-specific IgE (Chondrex, Redmond, WA, United States), cytokines (IL-4, IL-5, IL-13, IL-17A, and IL-6; BD Pharmingen, San Diego, CA, United States), and eotaxin (R&D Systems, Minneapolis, MN, United States) were analyzed using enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions. SCFA levels in the cecal supernatants were analyzed using mass spectrometry (EZmass Co., Ltd., Gyeongnam, Korea). Briefly, 50 mg of cecal content were mixed with 500 μL of distilled water and 10 μL of 5 M HCl, sonicated for 10 min, and then added 400 μL of ether. The mixture was centrifuged, and 200 μL of supernatants were derivatized by adding 20 μL N,O-bis(trimethylsilyl)-trifluoroacetamide (BSTFA; Machery-Nagel, Düren, Germany). One microliter of the sample was injected into a GC-2010 Plus instrument equipped with a triple quadrupole mass spectrometer TQ-8030 (Shimadzu, Kyoto, Japan).
Tq8030
Manufactured by Shimadzu
Sourced in Japan, Germany
The TQ8030 is a triple quadrupole mass spectrometer designed for high-performance liquid chromatography (HPLC) analysis. It offers precise and accurate quantitative measurements of compounds in complex matrices. The core function of the TQ8030 is to provide sensitive and selective detection of target analytes.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 4 (il 4), by BD (1 mentions)
Gc 2010 plus instrument, by Shimadzu (1 mentions)
Eotaxin, by R&D Systems (1 mentions)
Bstfa, by Macherey-Nagel (1 mentions)
Total ige, by BioLegend (1 mentions)
Hdm specific ige, by Chondrex (1 mentions)
Rtx 5ms, by Restek (1 mentions)
Slb 5ms, by Merck Group (1 mentions)
β glucuronidase arylsulfatase from helix pomatia, by Merck Group (1 mentions)
Sep pak c18 cartridge, by Waters Corporation (1 mentions)
Poroshell c18 column, by Agilent Technologies (1 mentions)
Ultra performance liquid chromatography (uplc), by Shimadzu (1 mentions)
8 protocols using tq8030
1
Quantification of Immune Markers and SCFAs
2
Untargeted GC-MS Metabolite Profiling of Aloe Extracts
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The metabolite profile of the Aloe extracts, including multiple batches of the same extract, was analyzed by using untargeted gas chromatography coupled mass spectrometry (GC-MS). Four out of five extracts, with multiple available batches were included in this analysis (Table 1 ). CX03 lacking multiple batches was not included in the metabolite analysis. Briefly, samples were extracted with water–methanol (1:9 v/v) containing ten stable isotope labeled internal standards followed by drying and derivatization by using oxymation and silylation. The derivatized extracts were injected into a GC-MS system (Shimadzu TQ8030) (Shimadzu Europa GmbH, Duisberg, Germany) and GC-MS scan data (50–750 m/z) was analyzed for targeted peak detection [54 (link)]. Peaks were identified based on a Matlab script and data was normalized based on the internal standard peak intensities [55 (link)].
3
GC-MS Analysis of Thyme Oil
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Thyme oil has been screened for the identification of bioactive components by using GC–MS triple quadrupole (GC–MS TQ8030, Shimadzu Corp., Japan). Gas chromatography attached with Restek column (0.25 mm, 30 m, Rxi-5 ms) which was operated using Q3 scan acquisition mode (start time 3 min, end time 10 min, scan speed 2500, start m/z 40 and end m/z 700). The ionization voltage was 70 eV. The sample was introduced via all-glass injector working in the split mode and helium was used as a carrier gas (flow rate of 1 ml/min keeping the split ratio of 10:1). Initially, column temperature was kept at 100 °C, then increased linearly at 3 °C/min up to 300 °C and held for 5 min. The temperature of injection port was 250 °C and the GC–MS/MS interface was maintained at 300 °C. The identification of components was accomplished by comparing retention time and fragmentation pattern, as well as with mass spectra in the NIST spectral library stored in the computer software of the GC–MS/MS43 (link).
4
GC-MS Analysis of Bioflocculant EPS
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For identification of EPS monomers of the bioflocculant, gas chromatography-mass spectrometry (GC/MS) was done after derivatization of the non-volatile sugar monomers to a compatible volatile state. Dried EPS residue was dissolved in 80 μl of 20 mg mL-1 methoxyamine hydrochloride in pyridine for 90 min for 30°C. This was further derivatized by adding 80 μl of N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) and incubated for 30 min/37°C. Chromatography was done with an EB-5MS column (Agilent, USA) attached in a triple quadruple GC/MS (TQ8030, Shimadzu, Japan) supported with GC/MS solution software (version 4). Helium was used as the carrier gas with a flow rate of 1.0 mL min-1. GC/MS analysis was carried out with a modified method (Roessner et al., 2000 (link)). Mass range (m/z) was selected from 45 to 800 for the entire analysis. The GC program was optimized for detection of derivatives and all analyses were carried out with the split ratio of 20:1.
5
Essential Oil Chemical Composition Analysis
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The analysis of the chemical constituents of the essential oil was carried out using a GCMS System (TQ8030 Shimadzu) in the Laboratory of Chemistry of Natural Products of the Federal University of São Carlos (UFSCar).
Separation was performed on a fused-silica capillary column RTX-5MS (30 m × 0.25 mm id, 0.25 μm film thickness, Restek) using ultra-high purity helium as a carrier gas at a flow rate of 3.0 mL/min. The mass spectrometer was operated in the electron impact mode (EI) at 70 eV, scanning at a range of 43-550 m/z. The ion source temperature was set at 230 °C. The separation data were analysed using the GCMS Real Time Analysis ® Software. The temperature was initially maintained at 60 °C for 3 min, followed by an increase of 3
Separation was performed on a fused-silica capillary column RTX-5MS (30 m × 0.25 mm id, 0.25 μm film thickness, Restek) using ultra-high purity helium as a carrier gas at a flow rate of 3.0 mL/min. The mass spectrometer was operated in the electron impact mode (EI) at 70 eV, scanning at a range of 43-550 m/z. The ion source temperature was set at 230 °C. The separation data were analysed using the GCMS Real Time Analysis ® Software. The temperature was initially maintained at 60 °C for 3 min, followed by an increase of 3
6
Quantification of Bisphenol A in Serum
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BPA determinations were conducted as reported by Kosarac et al. [71 (link)], with some minor modifications as reported in [72 (link),73 (link)]. Briefly, d16-BPA was added as an internal standard and samples were defatted with hexane and then extracted with dichloromethane. The extracts were purified in two successive SPE steps, one with a Florisil solid phase and one with a C18 solid phase. Deconjugation was carried out using β-Glucuronidase/Arylsulfatase from Helix pomatia (Sigma-Aldrich). After nitrogen drying, purified serum extracts were derivatised with N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) (Sigma-Aldrich). Derivatised samples were quantified using triple quadrupole mass spectrometer hyphenated with gas chromatography by means of GCMS TQ8030 (Shimadzu, Kyoto, Japan).
The gas chromatographic method was based on a SLB-5ms, 10 m × 0.1 mm, with a film thickness of 0.1 μm (Supelco, Milan, Italy). The GC oven program includes an initial phase of 1.5 min at 188 °C and two ramps: the first at 20 °C/min up to 260 °C, the second at 40 °C/min up to 320 °C. The injection temperature was set at 260 °C, and the linear velocity of the carrier gas (He) was 70 cm/s. The total analytical time was 8 min. Two acquisition channels were used, one in SCAN mode between m/z = 50 and m/z = 500 and one in MRM mode: 357.10 > 191.20 m/z for BPA and 370.50 > 73.10 m/z for the d16-BPA.
The gas chromatographic method was based on a SLB-5ms, 10 m × 0.1 mm, with a film thickness of 0.1 μm (Supelco, Milan, Italy). The GC oven program includes an initial phase of 1.5 min at 188 °C and two ramps: the first at 20 °C/min up to 260 °C, the second at 40 °C/min up to 320 °C. The injection temperature was set at 260 °C, and the linear velocity of the carrier gas (He) was 70 cm/s. The total analytical time was 8 min. Two acquisition channels were used, one in SCAN mode between m/z = 50 and m/z = 500 and one in MRM mode: 357.10 > 191.20 m/z for BPA and 370.50 > 73.10 m/z for the d16-BPA.
7
Quantification of Plant Hormones
8
Plasma Metabolomics Analysis Protocol
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Fasting baseline and 20-week plasma samples (n = 48 subjects) were used to understand potential metabolite associations with the intervention outcomes. Plasma metabolomics was measured using a previously validated gas chromatography-mass spectrometry (GC-MS) method [24] . In brief, 100 µL of plasma from fasting blood samples taken at baseline and at 20 weeks was extracted with 900 µL methanol:water (9:1 v/v), including a mix of internal standards, and a 200-µL aliquot evaporated and derivatized using methoxymation and silylation. The metabolic profile was measured using GC-MS (Shimadzu TQ8030, Shimadzu Europa GmbH, Duisberg, Germany), and metabolites identified based on retention index and mass spectral matching.
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