Urinary SECO, ED, and ENL were assayed by isotope dilution gas chromatography-mass spectrometry in the SIM mode (HP 6890 GC, HP 5973 MSD; Agilent Technologies, Palo Alto, CA, USA) as described previously [15 (link)]. All lignan measures were normalized to urinary creatinine levels to adjust for urine concentration. Stool samples were thawed, homogenized, and genomic DNA was extracted as described previously [19 (link)]. DNA concentrations and purity were determined using the NanoDrop 8000 Spectrophotometer (ThermoFisher Scientific) and gel electrophoresis. Working stocks were diluted in AE buffer (QIAGEN, Germantown, MD, USA) from genomic DNA and samples were stored at −20 °C until shipped for sequencing. Pooled in-lab designed quality control samples were included in triplicate to assess variation in library preparation and sequencing batches [20 (link)]. The V1–V3 region of the 16S rRNA gene was sequenced using the Illumina MiSeq platform to obtain 2 × 300 bp paired-end reads. Fecal bacterial DNA extraction did not meet quality control standards for 15 participants and 5 participants were excluded for higher baseline vs. post-intervention enterolignan excretion (post-preintervention difference ≤−10 nmol/mg creatinine). Therefore, the final sample sizes for the present analyses were 228 completing baseline assessments and 170 completing all 4 visits.
Hp 5973 msd
Manufactured by Agilent Technologies
Sourced in United States
The HP 5973 MSD is a mass spectrometry detector designed for use with gas chromatography (GC) systems. It is capable of performing electron ionization (EI) and chemical ionization (CI) analysis of a wide range of compounds. The HP 5973 MSD provides high sensitivity, resolution, and mass accuracy for the detection and identification of various chemical substances.
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Lab products found in correlation
Hp 5ms column, by Agilent Technologies (3 mentions)
Hp 6890 plus gas chromatograph, by Agilent Technologies (2 mentions)
Hp 6890n plus chromatograph, by Agilent Technologies (2 mentions)
Nanodrop 8000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Ae buffer, by Qiagen (1 mentions)
Miseq platform, by Illumina (1 mentions)
Hp6890 gc, by Agilent Technologies (1 mentions)
Db 5ms capillary column, by Agilent Technologies (1 mentions)
Tensor 27 ft ir spectrophotometer, by Bruker (1 mentions)
Hp 6890 series gc, by Agilent Technologies (1 mentions)
Nh2 semi prep column, by Waters Corporation (1 mentions)
Varian 9012 solvent delivery system, by Agilent Technologies (1 mentions)
Hp 7890a plus gas chromatograph, by Agilent Technologies (1 mentions)
Hp 7890a plus chromatograph, by Agilent Technologies (1 mentions)
Hp 5msi column, by Agilent Technologies (1 mentions)
Hp 5ms, by Agilent Technologies (1 mentions)
9 protocols using hp 5973 msd
1
Urinary Lignans and Gut Microbiome Analysis
2
Quantitative Sterol Analysis of Plant Roots
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About 200 mg of freeze dried powdered roots from various treatments were extracted separately for two times with 1 mL of CHCl3: MeOH (7:3) at room temperature. 20 μg of 5α-Cholestane was employed as the internal standard. The extract upon drying in a rotary evaporator was saponified with 1.5 mL of MeOH and KOH (20% (v/v)) at 80 °C for 1 h, aimed to hydrolyze the sterol esters. Following it, about 1.5 mL of MeOH and 4N HCl each was added and incubated at 80 °C for 1 h. Then, this mixture was extracted thrice with 4 mL of hexane. The sterols got separated to the hexane layer, which was further evaporated to dryness. The resulting powder was subjected to trimethylsilylation with pyridine and N, O-bis (trimethylsilyl) trifluoroacetamide + 1% trimethyl chlorosilane (1:1) for 90 min at 37 °C. Finally, GC-MS analysis was performed using a gas chromatograph (6890 A; Agilent Technologies) with a DB-5 (MS) capillary column (30 m x 30.25 mm, 0.25-mm film thickness) coupled with a mass spectrometer (HP 5973MSD). The reaction conditions were exactly similar to30 . The peak area ratios of molecular ions of the endogenous sterol and that of internal standard were considered to be the actual endogenous sterol levels.
3
Synthesis and Characterization of Organic Compounds
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All the solvents and reagents were commercially available from Sigma‐Aldrich, Fluorochem, Alfa Aesar, and Fisher Scientific, and were used directly without further purification. Melting points were measured with the Gallenkamp melting point apparatus. 1H and 13C nuclear magnetic resonance (NMR) spectra were obtained by a 400 MHz Bruker Spectrospin which was fitted with a Bruker SampleXpress autosampler system, and Topspin 7.1 was used for NMR spectra analysis. Chemical shifts of all compounds were calibrated with tetramethylsilane (TMS at δ = 0), with splittings outlined as singlet (s), doublet (d), and triplet (t). Fourier‐transform infrared (FTIR) spectroscopy was performed on Bruker TENSOR 27 FTIR spectrophotometer with the sample prepared using the KBr pellet press method. High‐resolution mass spectroscopy (HRMS) was performed using Agilent HP6890 GC with HP 7683 Injector interfaced directly to Agilent HP 5973 MSD (EI) instrument.
4
NPLC-UV and GC/MS Analysis of Compounds
5
GC and GC-MS Analysis of Compounds
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Gas chromatographic (GC) analysis was performed on an Agilent Technologies HP 7890A Plus Gas chromatograph equipped with a FID and fitted with HP-5ms column (30 m × 0.25 mm, film thickness 0.25 μm, Agilent Technologies, Santa Clara, CA, USA). The analytical conditions were: carrier gas H2 (1 mL/min), injector temperature (PTV: programmable temperature vaporization) 250 °C, detector temperature 260 °C, column temperature programmed from 60 °C (2 min hold) to 220 °C (10 min hold) at 4 °C/min. Samples were injected using a split mode with a split ratio of 10:1. The volume injected was 1.0 μL. Inlet pressure was 6.1 kPa.
An Agilent Technologies (Santa Clara, CA, USA) HP 7890A Plus Chromatograph fitted with a fused silica capillary HP-5ms column (30 m × 0.25 mm, film thickness 0.25 μm) and interfaced with a mass spectrometer HP 5973 MSD was used for the GC/MS analysis, under the same conditions as those used for GC analysis. The conditions were the same as described above with He (1 mL/min) as carrier gas. The MS conditions were as follows: ionization voltage 70 eV; emission current 40 mA; acquisitions scan mass range of 35–350 amu at a sampling rate of 1.0 scan/s. Compound identification was carried out by comparison of the MS fragmentation patterns and calculated retention indices with those available in the databases [35 ,36 ,37 ] and, when available, with standard substances.
An Agilent Technologies (Santa Clara, CA, USA) HP 7890A Plus Chromatograph fitted with a fused silica capillary HP-5ms column (30 m × 0.25 mm, film thickness 0.25 μm) and interfaced with a mass spectrometer HP 5973 MSD was used for the GC/MS analysis, under the same conditions as those used for GC analysis. The conditions were the same as described above with He (1 mL/min) as carrier gas. The MS conditions were as follows: ionization voltage 70 eV; emission current 40 mA; acquisitions scan mass range of 35–350 amu at a sampling rate of 1.0 scan/s. Compound identification was carried out by comparison of the MS fragmentation patterns and calculated retention indices with those available in the databases [35 ,36 ,37 ] and, when available, with standard substances.
6
GC and GC/MS Analysis of Samples
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Gas chromatography (GC) analysis was performed on an Agilent Technologies HP 6890 Plus Gas chromatograph equipped with a FID and fitted with HP-5MS column (30 m ×0.25 mm, film thickness 0.25 µm, Agilent Technology) . The analytical conditions were: carrier gas He (1 mL/min) , injector temperature (PTV) 250℃, detector temperature 260℃, column temperature programmed from 60℃ (2 min hold) to 220℃ (10 min hold) at 4 ℃/min. Samples were injected by splitting and the split ratio was 10:1. The volume injected was 1.0 µL. Inlet pressure was 6.1 kPa. The relative amounts of individual components were calculated based on the GC peak area (FID response) , as previously described 17, 18) .
An Agilent Technologies HP 6890N Plus Chromatograph fitted with a fused silica capillary HP-5 MS column (30 m× 0.25 mm, film thickness 0.25 µm) and interfaced with a mass spectrometer HP 5973 MSD was used for the gas chromatography-mass spectrometry (GC/MS analysis) The same conditions described above was also used for GC. The mass spectrum operating conditions were as follows: ionization voltage 70eV; emission current 40 mA; acquisitions scan mass range of 35-350 amu at a sampling rate of 1.0 scan/s.
An Agilent Technologies HP 6890N Plus Chromatograph fitted with a fused silica capillary HP-5 MS column (30 m× 0.25 mm, film thickness 0.25 µm) and interfaced with a mass spectrometer HP 5973 MSD was used for the gas chromatography-mass spectrometry (GC/MS analysis) The same conditions described above was also used for GC. The mass spectrum operating conditions were as follows: ionization voltage 70eV; emission current 40 mA; acquisitions scan mass range of 35-350 amu at a sampling rate of 1.0 scan/s.
7
Chemical Composition of A. galanga Essential Oil
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Fresh rhizomes of A. galanga were washed with clean water and were cut into small pieces before being subjected to hydro-distillation for 3 h. The obtained AGO was analyzed for their chemical compositions by gas chromatography–mass spectrometry (GC–MS) on an Agilent 6890 gas chromatograph coupled to electron impact (EI, 70 eV) using a Hewlett Packard (HP) mass selective detector (MSD), model HP 5973-MSD (Agilent Technologies Inc., Willmington, DE, USA). The HP5-MSI column with a 30.0 m × 0.25 mm internal diameter and a 0.25 mm film thickness (Agilent Technologies Inc., Santa Clara, CA, USA) was used as a capillary column. The analytical conditions were modified from previous studies [22 (link)]. Briefly, AGO was diluted with dichloromethane to 1:100 (v/v) and 1 μL of this mixture was injected into GC–MS. The injection and detector temperatures were 250 °C and 280 °C, respectively. The oven temperature was 70 °C. The sample was held isothermally for 3 min and the temperature was then increased by 3 °C/min to 188 °C, and then by 20 °C/min to 280 °C, followed by holding for 3 min. Helium was used as the carrier gas at a flow rate of 1 mL/min. The experiments were performed in triplicate.
8
GC and GC-MS Analysis of Essential Oils
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Gas chromatography (GC) analysis was performed on an Agilent Technologies HP 6890 Plus Gas chromatograph equipped with a FID and fitted with HP-5MS column (30 m x 0.25 mm, film thickness 0.25 m, Agilent Technology). The analytical conditions were: carrier gas H2 (1 mL/min), injector temperature (PTV) 250ºC, detector temperature 260ºC, column temperature programmed from 60ºC (2 min hold) to 220ºC (10 min hold) at 4 o C/min. Samples were injected by splitting and the split ratio was 10:1. The volume injected was 1.0 L. Inlet pressure was 6.1 kPa. Each analysis was performed in triplicate. The relative amounts of individual components were calculated based on the GC peak area (FID response).
An Agilent Technologies HP 6890N Plus Chromatograph fitted with a fused silica capillary HP-5 MS column (30 m x 0.25 mm, film thickness 0.25 m) and interfaced with a mass spectrometer HP 5973 MSD was used for the GC/MS analysis, under the same conditions as those used for GC analysis. The conditions were the same as described above with He (1 mL/min) as a carrier gas. The MS conditions were as follows: ionization voltage 70 eV; emission current 40 mA; acquisitions scan mass range of 35-350 amu at a sampling rate of 1.0 scan/s. The MS fragmentation patterns were checked with those of other essential oils of known composition.
An Agilent Technologies HP 6890N Plus Chromatograph fitted with a fused silica capillary HP-5 MS column (30 m x 0.25 mm, film thickness 0.25 m) and interfaced with a mass spectrometer HP 5973 MSD was used for the GC/MS analysis, under the same conditions as those used for GC analysis. The conditions were the same as described above with He (1 mL/min) as a carrier gas. The MS conditions were as follows: ionization voltage 70 eV; emission current 40 mA; acquisitions scan mass range of 35-350 amu at a sampling rate of 1.0 scan/s. The MS fragmentation patterns were checked with those of other essential oils of known composition.
9
GC/MS Analysis of Organic Extracts
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The chemical analysis of the organic extract was performed by GC/MS using an HP 6800 chromatograph (Agilent Technologies) coupled to a mass spectrometer HP 5973 MSD equipped with a capillary column HP-5MS (5% phenyl and 95% dimethyl polysiloxane) with dimensions of 30 m x 0.25 mm x 0.25 microns. The temperature in the column was programmed at 60 °C to 290 °C at 6 °C. min -1 and then held isothermal for 5 min. The gas carrier is helium and flow at a rate of 0.8 mL.min -1 . The injector temperature was at 250 °C, the volume of extract injected is 1 µl in split mode (20:1), and the solvent is hexane (within 4 min). The interface temperature was 280 °C, and of ionization source was 230 °C and the ionization was effected by electron impact (EI) with a potential of 70 eV. The analyzer was a quadrupole (150 °C). The mass spectrum is recorded using a mass detector scan mode (34-550 amu) . The GC/MS can detect compounds according to their elution order and identify by comparing their mass spectrum and retention time (RT) with those of Wiley databases NIST 7 and 02. The content of chemicals constituents is expressed by percentages peak-area (Area %). The compounds selected for identification are those with a recognition rate higher than 30%
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