Phenomenex Kinetex C8, 5 μm, 100 A, 100 mm × 2.1 mm maintained at 40 °C.
Kinetex c8
Manufactured by Phenomenex
Sourced in United States, Japan
The Kinetex C8 is a reversed-phase liquid chromatography (RPLC) column that utilizes a porous core-shell particle technology. It is designed for the separation and analysis of a wide range of compounds, including polar, non-polar, and moderately hydrophobic molecules. The Kinetex C8 column provides efficient and fast separations with low backpressure, making it suitable for use in both conventional and ultra-high-performance liquid chromatography (UHPLC) systems.
Automatically generated - may contain errors
Lab products found in correlation
Lcms 8060, by Shimadzu (4 mentions)
8050 triple quadrupole mass spectrometer, by Shimadzu (3 mentions)
Labsolutions software, by Shimadzu (2 mentions)
Nexera uhplc system, by Shimadzu (2 mentions)
Labsolutions insight, by Shimadzu (2 mentions)
Lc ms ms method package for phospholipid profiling, by Shimadzu (2 mentions)
Ultimate 3000 rslcnano system, by Thermo Fisher Scientific (2 mentions)
Lcms 8040, by Shimadzu (1 mentions)
Lcms 8050, by Shimadzu (1 mentions)
Nexera x2 uhplc system, by Shimadzu (1 mentions)
Lcms 8050 triple quadrupole mass spectrometer, by Shimadzu (1 mentions)
Uhplc, by Shimadzu (1 mentions)
Lcms 9030, by Shimadzu (1 mentions)
4000 qtrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Agilent 1260 lc system, by Agilent Technologies (1 mentions)
Gemini c18, by Phenomenex (1 mentions)
Securityguard guard column, by Phenomenex (1 mentions)
Chromnav2, by Jasco (1 mentions)
Integrated hplc system, by Jasco (1 mentions)
As 4050, by Jasco (1 mentions)
25 protocols using kinetex c8
1
Reversed-Phase Liquid Chromatography
2
Quantitative Lipid Profiling in MDCK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In all, 1.0 × 106 MDCK cells, MDCK-pTR GFP-RasV12 cells or 1:1 mix culture of MDCK and MDCK-pTR GFP-RasV12 cells were plated into non-coat plastic dishes (6-well, greiner) (triplicate under each condition). After incubation for 12 h, tetracycline was added to induce GFP-RasV12 expression, followed by incubation for 16 h. The culture media were then changed to foetal calf serum (FCS)-free Dulbecco’s modified Eagle’s medium with tetracycline, which were collected after 6 h and stored at −80 °C. After thawing, the samples were centrifuged for 10 min at 10,000 × g and were purified with solid phase extraction with an Oasis HLB cartridges (Milford, MA, USA) as previously described60 (link). Liquid chromatography–selected reaction monitoring–mass spectrometry (LC-SRM-MS) analysis was performed using a Nexera UHPLC system and a triple quadrupole mass spectrometer LCMS-8040 (Shimadzu, Kyoto, Japan) with a reversed-phase column (Kinetex C8, 2.1 × 150 mm, 2.6 µm; Phenomenex, Torrance, CA) as described previously61 (link). LabSolutions software (Shimadzu) was used for peak detection and integration. Chromatographic peak area for the corresponding SRM transitions were used for comparison of lipid mediators.
3
Quantitative PI Profiling by LCMS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipids were extracted from cell pellets through Bligh and Dyer method. Nexera UHPLC system equipped with LCMS-8050 (Shimadzu Corp., Kyoto, Japan) was used for chromatographic separation and detection of PIs. Separation was performed at 45 °C using an analytical column, Kinetex C8 (2.1 mm id × 150 mm length, 2.6 µm) (Phenomenex). The gradient mobile phase system consisted of 20 mM ammonium formate (mobile phase A) and 50% acetonitrile, 50% 2-propanol (mobile phase B). ESI-negative mode was used for the detection of PIs. PI(12:0/13:0) (Merck) was added as an internal standard for calculating PI concentrations.
4
Comprehensive Lipid Mediator Analysis by LC-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analyses were carried out with a Shimadzu Nexera X2-UHPLC system coupled to a Shimadzu LCMS-8050 triple quadrupole mass spectrometer. A reversed-phase column (Kinetex C8, 2.1 × 150 mm, 2.6 μm, Phenomenex) was used for chromatographic separation. Comprehensive analysis was performed by using LC–MS solution software and the LC–MS/MS Method Package for Lipid Mediators version 2 (Shimadzu). Metabolomics data were processed by using Excel software (Microsoft Corporation, Redmond, WA, USA). Missing values in the raw data were replaced by half of the minimum positive value, and these data were used for subsequent statistical analysis. The lipid mediator pathways were visualized by using VANTED software (https://www.cls.uni-konstanz.de/software/vanted/ )57 (link).
5
Comprehensive Lipid Mediator Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis of lipid mediators was performed as described in Simard et al. [28 (link),33 (link)]. Briefly, the dermis was reduced to a fine powder (as described in Section 2.5 ), which was then suspended in 500 μL Tris-hydrochloride 50 mM (pH 7) and immediately denatured in one volume of cold methanol containing the internal standard (Table S2 ). Lipids were extracted using an acidified methanol–chloroform method as described elsewhere [34 (link)]. The extracted lipids were reconstituted in 50 μL of a liquid chromatography solvent (Solvent A and B, 50/50) and 40 μL was injected onto a reversed-phase HPLC column (Kinetex C8, 150 × 2.1 mm, 2.6 μm; Phenomenex, Torrance, CA, USA) in a LC-MS/MS system [35 (link)]. Solvent A was composed of water containing 0.05% acetic acid and 1 mM ammonium cation, while solvent B was composed of acetonitrile with water (95/5, v/v), 0.05% acetic acid, and 1 mM ammonium cation. Finally, lipids were quantified using calibration curves generated with pure standards in triplicates.
6
Quantification of PF-05251749 in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PF‐05251749 samples were assayed using a validated high‐performance liquid chromatography tandem mass spectrometric (HPLC–MS/MS) method. Samples (50.0 μl) were extracted using liquid–liquid extraction (ethyl acetate) and extracts were evaporated to dryness followed by reconstitution in 500 μl of acetonitrile: water (1:1 v/v). Reconstituted extracts were centrifuged, and 10 μl of the supernatant was introduced for chromatographic separation on Phenomenex Kinetex C8, 2.6 μm, 30 × 2.1 mm, followed by precursor ion monitoring method (310.1 ➔ 211.0) in the positive ion mode of ionization. PF‐06810368 (314.1 ➔ 211.3) was used as the internal standard for this method. The between‐day assay accuracy, expressed as percent relative error (%RE), for quality control (QC) concentrations, ranged from −4.7 to −1.1%. Calibration standard responses were linear over the range of 1.00–1000 ng/ml using weighted (L/concentration2 ) linear least squares regression. Samples with concentrations above the upper limits of quantification were adequately diluted into calibration range. The lower limit of quantification (LLOQ) for PF‐05251749 was 1.00 ng/ml. Clinical specimens with plasma PF‐05251749 concentrations below the LLOQ were reported as below the LLOQ (<1.00 ng/ml).
7
LC-MS/MS Analysis of Small Molecules
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MS and MS/MS spectra were recorded on an LC/Q-TOF system consisting of a Nexera UHPLC and aquadrupole time-of-flight mass spectrometer (LCMS-9030,Shimadzu Corporation, Kyoto, Japan). LC conditions were as follows; column, Kinetex C8 (2.1 mmI.D. × 150 mm, 2.6 µm) Phenomenex, USA; solvent, isocratic, 30% of 20 mM ammonium formate-containing and 70% acetonitrile/isopropanol (1:1, v/v); flow rate, 0.4 mL/min; and column temperature, 40 °C. MS conditions were as follows: ionization mode, ESI positive/negative; capillary voltage, 4.5 kV (positive)/− 3.5 kV (negative); drying gas, 10 L/min; nebulizer gas, 2.0 L/min, heating gas, 10 L/min; interface temperature, 300 °C.
8
Quantification of Endogenous PAF and Lyso-PAF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Endogenous PAF and lyso-PAF were measured using a triple quadrupole mass spectrometer LCMS-8060 (Shimadzu, Japan) as previously described.25 (link) In brief, a reverse-phase column (Kinetex C8, 2.1 × 150 mm, 2.6 μm, Phenomenex) was used for chromatographic separation with a binary mobile phase of the following compositions: 0.1% formic acid/water (mobile phase A) and acetonitrile (mobile phase B). The gradient of the mobile phase (%A/%B) was programmed as follows: 0 min (90/10), 5 min (75/25), 10 min (65/35), 20 min (25/75), 20.1–28 min (5/95), 28.1–30 min (90/10). The flow rate was 0.4 mL/min, and the column temperature was 40°C. The selected reaction-monitoring transitions were: m/z 568.4 → 59.1 (PAF), m/z 572.4 → 59.1 (PAF-d4), m/z 482.3 → 104.2 (lyso-PAF), and m/z 486.3 → 104.2 (lyso-PAF-d4). Raw data were analyzed using LabSolutions Insight (Shimadzu), and the signals were compared with those of standard curves for quantification. The calculated data are listed in Table S1 .
9
LC-MS/MS Analysis of C-CTX-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Confirmation of C-CTX-1 in this study was carried out using a modified protocol [18 (link)]. The analysis was performed on an Agilent 1260 LC system (Agilent Inc., Palo Alto, California) coupled to a QTRAP 4000 mass spectrometer (Applied Biosystems, Inc., Foster City, California). The analyte was eluted on a Kinetex C8 (75 × 2.1 mm; 2.6 μm) column (Phenomenex, Torrance, California) using a mobile phase consisting of A (water) and B (95% aqueous acetonitrile), both containing 0.1% formic acid. The mobile phase program consisted of 10% B for 1 min, then to 95% B at 1.5 min, held at 95% B for 5 min, and returned to 10% B in 0.2 min. The column was re-equilibrated with 10% B for 2.8 min. The flow rate was set to 0.3 mL min-1 and the column temperature and auto-injector tray were maintained at 40°C and 10°C, respectively. The injection volume was set at 5 μL. The mass spectrometer was operated in positive electrospray ionization mode. Principal instrument settings maintained were: CUR 20 psi, IS 5500 V, TEM 400°C, GS1and GS2 both at 60 psi; CAD medium; DP: 75 V, EP 10 V, CE 35 eV and CXP 15 V. Analyst 1.6.1 was used for data acquisition. C-CTX-1 standard used in the LC-MS/MS analysis was provided by the FDA.
10
Lipid Extraction and eCBome Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipids were extracted from tissue samples according to the Bligh and Dyer method [94 (link)]. About 10 mg of each tissue were homogenized in 1 mL of Tris-HCl 50 mM pH 7 and following 1 mL of methanol using a pestle. 5 μL of eCBome deuterated standards and 5.75 μL of 0.1 M acetic acid were added to each tube and then 1 mL of chloroform was added to each sample, vortexed and centrifuged at 3000× g for 5 min. The chloroform step was repeated two more times for a total addition of 3 mL of chloroform. The lipid phases were collected and dried by stream of nitrogen and then diluted in 50 μL of mobile phase containing 50% of solvent A (water + 1 mM ammonium acetate + 0.05% acetic acid) and 50% of solvent B (acetonitrile:water 95:5 + 1 mM ammonium acetate + 0.05% acetic acid). Finally, diluted samples were injected onto an HPLC column (Kinetex C8, 150 × 2.1 mm, 2.6 μm, Phenomenex; Torrance, CA, USA) and eluted at a flow rate of 400 μL/min using a discontinuous gradient of solvent A and solvent B [95 (link)]. Afterwards, eCBome-related mediators were quantified by HPLC system interfaced with the electrospray source of a Shimadzu 8050 triple quadrupole mass spectrometer and using multiple reaction monitoring in positive ion mode for the compounds and their deuterated homologs.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!