The most active fungal extracts were selected to undergo secondary metabolites identification using gas-chromatography mass spectrometry GCMS, (GC-MS TQ8050; Shimadzu, Johannesburg, South Africa) equipped with a Multifunctional Autosampler (AOC-6000), a capillary column (RTX-5, 60 m × 0.25 mm × 0.25 µm, New Delhi, India) as described by Sharma et al. [7 ]. The identities of the compounds were determined by searching known molecules in databases of NIST05; WILEY 8, and FFNSC1.3 libraries.
Aoc 6000
Manufactured by Shimadzu
Sourced in Japan
The AOC-6000 is an automated sample injection system developed by Shimadzu. It is designed to automate the injection process for gas chromatography (GC) and liquid chromatography (LC) analyses, improving efficiency and repeatability.
Automatically generated - may contain errors
Lab products found in correlation
Gcms tq8050, by Shimadzu (1 mentions)
Rtx 5, by Shimadzu (1 mentions)
Tq8040 ms, by Shimadzu (1 mentions)
6495 triple quadrupole, by Agilent Technologies (1 mentions)
Nexis 2030, by Shimadzu (1 mentions)
Masshunter workstation b 07, by Agilent Technologies (1 mentions)
Zorbax eclipse plus c18 column, by Agilent Technologies (1 mentions)
Qp2010 ultra mass spectrometer, by Shimadzu (1 mentions)
Acquity uplc hss c18 column, by Waters Corporation (1 mentions)
Ilab 650, by Werfen (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Qp2010 plus, by Shimadzu (1 mentions)
Gc 2010 plus, by Shimadzu (1 mentions)
Gc 2010, by Shimadzu (1 mentions)
Gcms tq8040, by Shimadzu (1 mentions)
Car pdms, by Merck Group (1 mentions)
7 protocols using aoc 6000
1
Fungal Metabolite Identification by GC-MS
2
Essential Oil Analysis of Nepeta cataria
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Essential oil samples were prepared by the extraction of 10 µL of crude N. cataria essential oil with 1.5 ml of MTBE, which was then dried with Na2SO4 and centrifuged at 13 Krpm. The supernatant was transferred to a sampling vial for analysis. Essential oil separation was performed on a Shimadzu 2010 Plus gas chromatograph equipped with an AOC-6000 auto-sampler. The analysis of the relative abundance of compound fragments was performed on a Shimadzu TQ8040 MS.
An injection volume of 1 µL was separated using chromatographic grade helium on a H-Rxi-5Sil MS column held at 35 °C for 4 min then heated to 250 °C at 20 °C/min then held for 1.25 min at 250 °C. The inlet temperature was 250 °C with a splitless injection. The ion source temperature was set at 200 °C, the interface temperature was set at 250 °C, the solvent cut time was 3.5 min, and the detector voltage was set to 0.2 kV with a threshold of 1000. Peak integration percentages were generated using the GCMSsolution v4.3© software from Shimadzu Corporation. Individual identities were determined by comparing the mass spectral results to current literature and screening them in the NIST05.LIB, NIST05s.LIB, W10N14.lib and the W10N14R.lib mass spectral libraries.
An injection volume of 1 µL was separated using chromatographic grade helium on a H-Rxi-5Sil MS column held at 35 °C for 4 min then heated to 250 °C at 20 °C/min then held for 1.25 min at 250 °C. The inlet temperature was 250 °C with a splitless injection. The ion source temperature was set at 200 °C, the interface temperature was set at 250 °C, the solvent cut time was 3.5 min, and the detector voltage was set to 0.2 kV with a threshold of 1000. Peak integration percentages were generated using the GCMSsolution v4.3© software from Shimadzu Corporation. Individual identities were determined by comparing the mass spectral results to current literature and screening them in the NIST05.LIB, NIST05s.LIB, W10N14.lib and the W10N14R.lib mass spectral libraries.
3
Comprehensive GC-MS/MS and UHPLC-MS/MS Analysis
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A Shimadzu Nexis 2030 gas chromatograph equipped with a programmed split/splitless injector and an AOC 6000 multifunction autosampler (Shimadzu, Kyoto, Japan), and a Shimadzu 8040 138 NX tandem mass spectrometry (Shimadzu, Kyoto, Japan) were used to perform gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) confirmation. An SH-Rxi-5Sil MS (30 m 140 × 0.25 mm id × 0.25 μm film) capillary column was used. A 1290 Infinity UHPLC system was linked to a 6495 Triple Quadrupole LC-MS/MS device added with a jet stream EI source (Agilent, Santa Clara, CA, USA). Data were acquired and analyzed on an Agilent MassHunter Workstation B.07.00. Chromatographic isolation was finished on an Agilent ZORBAX Eclipse Plus C18 column (50 mm × 2.1 mm, 1.8 μm) with gradient elution.
Samples were prepared using a GENIUS 3 vortex agitator (IKA, Stauffen, Germany), a CL31R multispeed refrigerated centrifuge (Thermo Scientific, Waltham, MA, USA), a WD12 water bath nitrogen blowing instrument (Aosheng Instrument, Hangzhou, China), and a CK2000 high-throughput tissue grinder (Thmorgan Biotechnology, Beijing, China).
Samples were prepared using a GENIUS 3 vortex agitator (IKA, Stauffen, Germany), a CL31R multispeed refrigerated centrifuge (Thermo Scientific, Waltham, MA, USA), a WD12 water bath nitrogen blowing instrument (Aosheng Instrument, Hangzhou, China), and a CK2000 high-throughput tissue grinder (Thmorgan Biotechnology, Beijing, China).
4
Comprehensive Toxicological Analysis of Biological Samples
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For each case, a comprehensive toxicological analysis, including general screening and quantification of drugs of abuse and medicinal drugs, was performed. Particularly, analyses for alcohol were done by gas chromatography coupled to a Flame Ionization Detector (Shimadzu QP 2010 Plus, Kyoto, Japan). Blood samples were initially screened for illicit drugs (cannabinoids, cocaine, opiates, methadone, and amphetamines/methamphetamines/MDMA/MDA) by immunoassay (ILab 650, Werfen, Barcelona, Spain) [19 (link)]. Confirmation analyses for cannabinoids were performed with a Shimadzu GC-2010 Plus gas chromatograph equipped with a model AOC-6000 auto-sampler system and interfaced with a QP 2010 Ultra mass spectrometer (Shimadzu, Kyoto, Japan) using a previously validated method [20 (link)]. Confirmation analyses for other illicit drugs and screening/confirmation for 68 psychoactive medications (benzodiazepines, Z-drugs, antipsychotics, antidepressants, and medical opioids) were performed with an ACQUITY UPLC® System (Waters Corporation, Milford, MA, USA) equipped with an Acquity UPLC® HSS C18 column (2.1 × 150 mm, 1.8 μm; Waters) using a previously validated method [21 (link),22 (link)].
5
Hexanal Quantification via SPME-GC-FID
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Headspace hexanal measurements were performed via solid-phase microextraction coupled to a gas chromatograph with a flame ionization detector (SPME-GC-FID) using a Shimadzu GC-2010 with a Shimadzu AOC-6000 autosampler (Shimadzu, Kyoto, Japan) [43 (link)]. GC vials were heated in an autosampler at 55 °C for 10 min. After heating, the volatiles were absorbed on the surface of a 50/30 mm divinylbenzene/carboxen/polydimethylsiloxane SPME fiber needle (Supelco, Bellefonte, PA, USA), which was inserted for 2 min into the GC vials at the end of the heating process. The fiber needle carrying the volatile compounds was placed into the injector port of the GC where they are desorbed at 250 °C for 3 min and separated on a 30 m × 0.32 mm i.d. × 1 μm fused silica capillary Equity-1 column for 10 min using helium as the carrier gas. The oven temperature was 65 °C, while the FID was at 250 °C and a split ratio of 1:7 was used. The area under the curve was used for quantification using a standard curve prepared with (0–200 μM) authentic hexanal.
6
Volatile Compound Extraction from Cookies
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Volatile compounds of cookie samples were extracted by the method of HS-SPME (AOC-6000, Shimadzu, Kyoto, Japan). 2.0 g of ground cookie powder was placed into 20 mL headspace vials and balanced at 60 °C for 30 min. A 2 cm CAR/PDMS/DVB (50/30 μm) SPME fiber (Supelco, Bellefonte, PA, USA) was inserted into a sample bottle for 20 min. After that, the fiber was desorbed in the gas chromatography (GC) injector at 250 °C for 3 min.
7
Volatile Compounds Analysis of Maca Powders
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Maca powders (1.0 g) of each sample were placed in a 20-mL headspace bottle, and sealed immediately with a silicon lid. After equilibrium in an heated oscillator at 65 °C for 10 min, the volatiles were extracted at 350 rpm for 30 min and placed at the GC injection port for 2 min for desorption. The mass spectrometer (GCMS-TQ8040, Shimadzu, Tokyo, Japan) was equipped with an autosampler AOC-6000 and a InertCapPure-Wax column (30 m × 0.25 mm × 0.25 µm). Solid phase extraction was conducted with the fibers 85 μm PA, 95 μm CAR WR/PDMS, and 75 μm CAR/PDMS (Supelco). Helium (>99.99%) was used as the carrier gas at a constant flow rate of 1 mL/min. The injection volume was 1 mL in splitless mode. The initial oven temperature was held at 50 °C for 5 min, ramped to 250 °C at a rate of 10 °C/ min, and held at 250 °C for 10 min. The temperatures of injector, pressure, split ratio, and electron impact ion source were set to 250 °C, 83.5 kPa, 5:1, and 200 °C, respectively. The qualitative and semi-quantitative data was collected in a full scan mode (m/z 45–450) and multiple reaction monitoring (MRM), respectively.
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