Analyses were performed with a HP6890 gas chromatograph (Hewlett-Packard, Roma, Italia) combined with a 7683 series Injector (Agilent, Milan, Italy) coupled with a HP 5973 quadrupole mass spectrometer (Hewlett-Packard, Roma, Italia).
Hp 6890 gas chromatograph
Manufactured by Hewlett-Packard
Sourced in United States
The HP 6890 gas chromatograph is a laboratory instrument designed for the separation, identification, and quantification of chemical compounds in complex mixtures. It utilizes an inert carrier gas, such as helium or nitrogen, to transport the sample through a column filled with a stationary phase material. The analytes in the sample are separated based on their different interactions with the stationary phase, allowing for their individual detection and measurement.
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Lab products found in correlation
Hp 5ms capillary column, by Agilent Technologies (2 mentions)
Hp 5973, by Hewlett-Packard (2 mentions)
7683 series injector, by Agilent Technologies (1 mentions)
Sp 2330 fused silica capillary column, by Merck Group (1 mentions)
Ac 400 spectrometer, by Bruker (1 mentions)
Hp 5ms column, by Hewlett-Packard (1 mentions)
Esquire 3000 plus spectrometer, by Bruker (1 mentions)
Hp 5ms, by Hewlett-Packard (1 mentions)
Db 5ms column, by Hewlett-Packard (1 mentions)
Agilent 7890a gas chromatograph, by Agilent Technologies (1 mentions)
Agilent 5975c mass spectrometric detector, by Agilent Technologies (1 mentions)
Kjeltec system, by Foss (1 mentions)
Du530 spectrophotometer, by Beckman Coulter (1 mentions)
Hp ffap capillary column, by Agilent Technologies (1 mentions)
Digitheat, by J.P. Selecta (1 mentions)
Db 23 capillary column, by Merck Group (1 mentions)
10 protocols using hp 6890 gas chromatograph
1
Gas Chromatography-Mass Spectrometry Analysis
2
Fatty Acid Profiling in Feed and Liver
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Total lipids were extracted from approximately 2 g of feed or liver tissue samples using a 2:1 solution of chloroform and methanol [16 (link)]. Fatty acid methyl esters were prepared with boron trifluoride methanol as the methylating agent using methods reported earlier [15 (link)]. Fatty acid analysis was performed with a HP 6890 gas chromatograph (Hewlett-Packard Co., Wilmington, DE, USA) equipped with an autosampler, flame ionization detector, and SP-2330 fused silica capillary column (30 m × 0.25 mm × 0.2 μm film thickness) (Supelco, Bellefonte, PA, USA). Samples in hexane (1 μL) were injected with helium as a carrier gas into the column programmed for ramped oven temperatures. Initial oven temperature was set at 150 °C, held for 1.5 min, then ramped at 15 °C/min to 190 °C and held for 20 min, then ramped again at 30 °C/min to 230 °C and held for 3 min. Inlet and detector temperatures were both 250 °C. Fatty acid methyl esters were identified by comparison with retention times of authentic standards (Catalog no:1177, Matreya LLC, State College, PA, USA). Peak areas and percentages were calculated using Hewlett-Packard ChemStation software (Agilent Technologies Inc., Wilmington, DE, USA). Fatty acid values are reported as mg/g tissue and g/100 g (%) of FA methyl esters.
3
Sterol Profiling of Oils by GC-FID
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The qualitative and quantitative sterol contents of the samples were determined according to the European Official Methods as described by Sanchez-Casas et al. 22) . The lipid, after addition of α-cholestanol and betulin as internal standards, was saponified with ethanolic potassium hydroxide solution. The unsaponifiable matter was extracted with diethyl ether. The sterol fraction was separated by Silica gel plate chromatography. The sterols, erythrodiol and uvaol, recovered from the plate were transformed into the corresponding trimethylsilyl ethers and the mixture was analysed by gas chromatography using an HP 6890 gas chromatograph (Hewlett-Packard, Agilent, CA) , equipped with a flame ionization detector (FID) , a HP-5MS capillary column (30 m×0.25 mm×0.25 μm) and a 6890 Agilent au-tomatic injector. The working conditions were: injector 300℃, isothermal analysis at 260℃, detector temperature 325℃. Quantification was performed by the internal standard (α-cholestanol) method and results were expressed as mg kg -1 of oil.
4
Fatty Acid Profile of Nigella sativa Oil
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According to the American Oil Chemists Society’s (AOCS) standard procedures, the peroxide value (PV), acid value (AV), saponification value (SV), and iodine value (IV) were assessed (American Oil Chemists' Society, 2005 ; American Oil Chemists' Society, 2011 ). Fatty acid methyl esters of N. sativa oil were prepared using a modified trans-methylation method according to AOAC (2016) , Official Methods of Analysis 2017. Determination of fatty acids composition was carried out using a Hewlett Packard HP 6890 gas chromatograph, operated under the following conditions: Detector, flame ionization (FID); column, capillary, 30.0 m × 530 μm, 1.0 μm thickness, polyethylene glycol phase (INNO Wax); N2 with flow rate, 15 ml/min with average velocity 89 cm/s (8.2 psi); H2 flow rate, 30 ml/min; air flow rate, 300 ml/min; split ratio, 8:1, split flow, 120 ml/min; gas saver, 20 ml/min. Detector temperature, 280°C; column temperature, 240°C; and injection temperature, 280°C. Programmed temperature starting from 100°C to reach a maximum of 240°C was used for eluting the fatty acid methyl esters. The identification of the peaks was made as compared with chromatograms of authentic standard fatty acids methyl esters mixture purchased from Sigma (United States).
5
Comprehensive NMR and Mass Spectroscopy Analysis
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1 H and 13 C-NMR spectra and DEPT experiments (Supplementary Materials): CDCl 3 solutions at rt using a Bruker-AC-400 spectrometer (Billerica, MA, USA) at 400, 100 and 100 MHz, respectively; 13 C spectra are proton decoupled; chemical shifts in ppm rel to internal SiMe 4 (=0 ppm).
TLC: Merk silica gel 60 F 254 plates (Merck Millipore, Milan, Italy). Column chromatography: silica gel.
Melting points were measured on a Reichert apparatus, equipped with a Reichert microscope, and are uncorrected.
Optical rotations were measured on a Jasco-DIP-181 digital polarimeter (Jasco, Tokyo, Japan).
Mass spectra were recorded on a Bruker ESQUIRE 3000 PLUS spectrometer (Electrospray Ionization (ESI) detector, Billerica, MA, USA or by GC-MS analyses.
GC-MS analyses: HP-6890 gas chromatograph equipped with a 5973 mass detector, using a HP-5MS column (30 m × 0.25 mm, 0.25 µm film thickness; Hewlett Packard, Palo Alto, CA, USA) with the following temp. program: 60 • (1 min), then 6 • /min to 150 • (held 1 min), then 12
TLC: Merk silica gel 60 F 254 plates (Merck Millipore, Milan, Italy). Column chromatography: silica gel.
Melting points were measured on a Reichert apparatus, equipped with a Reichert microscope, and are uncorrected.
Optical rotations were measured on a Jasco-DIP-181 digital polarimeter (Jasco, Tokyo, Japan).
Mass spectra were recorded on a Bruker ESQUIRE 3000 PLUS spectrometer (Electrospray Ionization (ESI) detector, Billerica, MA, USA or by GC-MS analyses.
GC-MS analyses: HP-6890 gas chromatograph equipped with a 5973 mass detector, using a HP-5MS column (30 m × 0.25 mm, 0.25 µm film thickness; Hewlett Packard, Palo Alto, CA, USA) with the following temp. program: 60 • (1 min), then 6 • /min to 150 • (held 1 min), then 12
6
Gas Chromatography and Mass Spectrometry Analysis
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Gas chromatography (GC) analyses were performed on a Hewlett-Packard (HP 6890) gas chromatograph (FID), equipped with a 5% phenyl methyl silicone HP-5 capillary column (30 m × 0.25 mm × film thickness 0.25 μm). The temperature was programmed from 50°C after 5 min initial hold to 200°C at 4°C/min. Gas c hromatography conditions were as follows: N2 as carrier gas (1.8 ml/min); split mode was used (Flow: 72.1 ml/min, ratio: 1/50); temperature of injector and detector was 250°C, Fin al hold time was 48 min. The machine was led by a computer system type ″HP ChemStation″, managing the functioning of the machine and allowing to follow the evolution of chromatographic analyses. Diluted samples (1/20 in methanol) of 1 μl were injected manually.
GC/MS qualitative analyses were performed on a Hewlett-Packard equipped with a HP-5MS (Crosslinked 5% PHME Siloxane) capillary column (30 m × 0.25 mm i.d, 0.25 μm film thickness) and coupled with a mass spectrometer (HP 5973). The temperature was programmed from 50 to 250°C at 2°C/min. The carrier gas was He (1.5 ml/min) and split mode was used (Flow: 112 ml/min, ratio: 1/74.7). The different compounds were confirmed by reference to their MS identities (Library of NIST/EPA/NIH MASS SPECTRAL LIBRARY Version 2.0, build Jul 1 2002). MS operating parameters were: ionization voltages 70eV, ion source temperature 230°C, scan mass range 35–450 amu.
GC/MS qualitative analyses were performed on a Hewlett-Packard equipped with a HP-5MS (Crosslinked 5% PHME Siloxane) capillary column (30 m × 0.25 mm i.d, 0.25 μm film thickness) and coupled with a mass spectrometer (HP 5973). The temperature was programmed from 50 to 250°C at 2°C/min. The carrier gas was He (1.5 ml/min) and split mode was used (Flow: 112 ml/min, ratio: 1/74.7). The different compounds were confirmed by reference to their MS identities (Library of NIST/EPA/NIH MASS SPECTRAL LIBRARY Version 2.0, build Jul 1 2002). MS operating parameters were: ionization voltages 70eV, ion source temperature 230°C, scan mass range 35–450 amu.
7
Quantitative Analysis of Analytes
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A screw-capped 10-ml glass extraction tube was used to collect 2 ml analyte, 100 μl IS (300 ng/mL), and 200 μl 12 NHCl solution. The extraction tube was then autoclaved (15 Psi, 120°C, 30 min). After cooling, a 1.5 ml phosphate buffer was added and mixed thoroughly. The sample was then adjusted to the indicated pH, followed by SPE. The organic was then transferred and evaporated under a stream of nitrogen. Once the samples were completely dried, 100 μL BSTFA and Ethyl acetate-mix (1:1) were added. The extraction tube was then capped and incubated at 100°C for 30 min. Samples were cooled to room temperature and 2 μL of derivatized samples were analyzed by a Hewlett-Packard (Palo Alto, CA, USA) HP 6890 gas chromatograph interfaced to an HP 5973 mass selective detector equipped with a DB-5MS column (30-M, 0.25-mm ID, 0.25-μm film thickness) to acquire full-scan and selected ion monitoring (SIM) mass spectrometric data. SIM mode was applied to quantify analytes [Table 1].
8
Lipid Isolation and Characterization
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Lipid isolation from the developed product was carried out by chloroform/methanol extraction according to the Folch method. The purity of the isolated lipids was tested using thin-layer chromatography. Determination of the fatty acid composition was carried out on an HP 6890 gas chromatograph (Hewlett Packard, Palo Alto, CA, USA), and fatty acids were identified by comparison with the software database. Cholesterol determination was performed on an Agilent 7890A gas chromatograph (Agilent Technologies, Santa Clara, CA, USA) with an Agilent 5975C mass spectrometric detector (Agilent Technologies, USA), an HP5MS capillary column (Agilent Technologies, USA) measuring 30 mm × 0.25 mm × 0.25 µm and helium as the carrier gas. Determination of the fatty acid composition and cholesterol content was performed according to the reported method in the literature [28 (link)].
Classic indices such as ΣSFA, ΣMUFA, ΣPUFA, Σn–6 PUFA and Σn–3 PUFA were calculated, and the PUFA/SFA, index of atherogenicity (IA), the hypocholesterolemic/hypercholesterolemic ratio (HH) and the unsaturation index (UI) were calculated according to the following equations [29 (link)]:
Classic indices such as ΣSFA, ΣMUFA, ΣPUFA, Σn–6 PUFA and Σn–3 PUFA were calculated, and the PUFA/SFA, index of atherogenicity (IA), the hypocholesterolemic/hypercholesterolemic ratio (HH) and the unsaturation index (UI) were calculated according to the following equations [29 (link)]:
9
Malt Analysis and Characterization
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The malts were finely ground into flour (2.0 mm) using a DLFU disc mill (Bühner-Miag, Braunschweig, Germany) according to method 3.1.4.2.1 [44 ]. In malt flour, the following parameters were determined: the moisture content by the drying oven method using a dryer Digitheat (J.P. Selecta, Barcelona, Spain) according to method 4.2 [41 ], the total nitrogen (TN) content by the Kjeldahl method using a Kjeltec System (Foss Tecator, Hoganas, Sweden) and converted into protein content (N × 6.25) according to method 3.1.4.5.1.1 [41 ], the diastatic power (DP) by the iodometric titration method [43 ] and the beta-glucan content by the spectrophotometric method after prior enzymatic hydrolysis using the (1-3) (1-4) Beta-D-Glucan Kit (Megazyme, Bray, Wicklow, Ireland) on a Beckman DU-530 spectrophotometer (Beckman, Wycombe, UK) according to method 4.16.1 [41 ]. The content of dimethyl sulfide precursors (DMS-P) in malts was determined by gas chromatography with flame photometric detection (GC/FPD) using an HP 6890 gas chromatograph (Hewlett Packard, Wilmington, Germany) with an HP 7694E headspace autosampler (Hewlett Packard, Wilmington, Germany) and an HP-FFAP capillary column (30 m × 0.53 mm × 1 µm) (Agilent Technologies, Santa Clara, CA, USA) according to method 3.1.4.17 [44 ]. The determinations were carried out in duplicate.
10
Soxhlet Extraction and GC Analysis of Fatty Acids
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An automated Soxhlet apparatus (Soctec Avanti 2055, Foss, Sweden) was used to extract the fat from MNM as described by the Association of Official Analytical Chemists (AOAC, 2006 , method number 920.39). The methyl ethers were analyzed using gas chromatography as described by Christopherson and Glass, (1969) . Briefly, the fat extracts were trans-methylated with 2 M methanol-sodium hydroxide. Heptane was used to extract the fatty acid methyl esters which were then filtered and dried under nitrogen. The fatty acids were separated by a temperature gradient over 45 min on gas chromatography with nitrogen as carrier gas on a DB-23 capillary column (90 cm × 250 μm × 0.25 μm) (Supelco, Sigma-Aldrich). The chromatograph consisted of an HP6890 gas chromatograph (Hewlett Packard, Bristol, United Kingdom) with a flame ionization detector (FID). Both the detector and injector temperatures were set at 300 °C. A PC equipped with Chemstation software was used for quantification. Nonadecanoic acid (C19:0) was used as an internal standard.
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