NMR spectra were recorded on ARX-400 Advance spectrometer (Bruker Corporation). FTIR spectra were obtained on a Nicolet iS10 infrared spectrometer (Thermo Scientific, Madison, WI) in reflection geometry using a single bounce diamond attenuated total reflectance (ATR) accessory. LPA were separated on a Luna™ C-8 (50 × 2 mm, 3 μm) column connected to a guard cartridge with 2.0 to 3.0 mm internal diameters (Phenomenex, Torrance, CA) in an Accela™ UHPLC system (Thermo Scientific, Madison, WI). MS data were collected via an LTQ-Orbitrap™ XL Discovery instrument (Thermo Scientific, Madison, WI), equipped with an ESI ion max source. All chemicals were purchased from Sigma-Aldrich (St. Louis, MO) and used without further purification except when specifically mentioned. All phospholipids were purchased from Avanti Polar Lipids (Alabaster, AL). Ethylene glycol dimethacrylate (EGDMA) was purified by vacuum distillation. Human plasma was collected by Lampire Biological Laboratories Inc., from female donors, processed to obtain platelet-free plasma, and frozen at −80 °C. Empty SPE tubes and frits were purchased from Sigma-Aldrich (St. Louis, MO). HPLC grade MeOH, CHCl3 and water were purchased from Honeywell International Inc.
Luna c8
Manufactured by Phenomenex
Sourced in United States
The Luna C8 is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of analytes. It features a silica-based stationary phase with octyl (C8) functional groups, providing a moderate level of hydrophobicity for the retention of moderately polar to nonpolar compounds. The Luna C8 column is available in various dimensions and particle sizes to accommodate different analytical needs.
Automatically generated - may contain errors
Lab products found in correlation
Luna c18, by Phenomenex (2 mentions)
Zorbax sb c18, by Agilent Technologies (2 mentions)
Ltq orbitrap xl discovery, by Thermo Fisher Scientific (1 mentions)
Nicolet is10 infrared spectrometer, by Thermo Fisher Scientific (1 mentions)
Accela uhplc system, by Thermo Fisher Scientific (1 mentions)
Api 3200, by AB Sciex (1 mentions)
Series 200 hplc system, by PerkinElmer (1 mentions)
Agilent 1100 1200 series, by Agilent Technologies (1 mentions)
System gold hplc, by Beckman Coulter (1 mentions)
Abi 4000 qtrap, by AB Sciex (1 mentions)
Lc 2010a system, by Shimadzu (1 mentions)
Scl 10a system, by Shimadzu (1 mentions)
C18 sphereclone, by Phenomenex (1 mentions)
1100 hplc system, by Agilent Technologies (1 mentions)
Alliance hplc, by Waters Corporation (1 mentions)
996 pda detector, by Waters Corporation (1 mentions)
Q tof mass spectrometer, by Waters Corporation (1 mentions)
Totalchrom workstation, by PerkinElmer (1 mentions)
Hypersil bds c8, by Thermo Fisher Scientific (1 mentions)
Series 200, by PerkinElmer (1 mentions)
17 protocols using luna c8
1
Comprehensive Analytical Workflow for Lipid Profiling
2
LC-MS/MS Quantification of Compound 12
3
LC-MS/MS Method for Methanol Extraction
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Methanol (MeOH) was used as extracting solvent. For washing, strips, viable epidermis and dermis, 17, 3, 2 and 5 ml were used, respectively. Viable epidermis and dermis were extracted with back and forth agitation for 3 hours. Washing extract and receptor fluid was diluted tenfold and twofold with MeOH, respectively. All samples were filtered on Millex filter 0.45 µm (Merck Millipore, Burlington, MA, USA) before analysis.
All samples were analysed onto an LC/MS-MS system (Agilent HP1200 LC system coupled with a mass spectrometer API 3200 (Sciex, Concord, Ontario, Canada). The analytical system was managed by the software Analyst version 1.6.
The analytical column used was a Luna C8 from Phenomenex (Torrance, CA, USA) (30 × 2.1 mm, 5 µm) and analysis were carried out with a gradient elution with mobile phases of 0.1% formic acid (FA) in water (A) and MeOH (B). The column temperature was fixed at 50 °C, the volume of the injection was 10 µl and the flow rate at 0.8 ml/min. Ionization mode used was APCI positive. Multiple Reaction Monitoring (MRM) was used for detection with the following transition 566.6 → 266.4.
LLOQ and ULOQ were at 2 and 2500 ng/ml, respectively. QCs used for extraction recovery and matrix effect was set at 20 and 500 ng/ml at least in triplicate. Extraction recovery for each analyzed compartment is reported in Supplementary TableS6 .
All samples were analysed onto an LC/MS-MS system (Agilent HP1200 LC system coupled with a mass spectrometer API 3200 (Sciex, Concord, Ontario, Canada). The analytical system was managed by the software Analyst version 1.6.
The analytical column used was a Luna C8 from Phenomenex (Torrance, CA, USA) (30 × 2.1 mm, 5 µm) and analysis were carried out with a gradient elution with mobile phases of 0.1% formic acid (FA) in water (A) and MeOH (B). The column temperature was fixed at 50 °C, the volume of the injection was 10 µl and the flow rate at 0.8 ml/min. Ionization mode used was APCI positive. Multiple Reaction Monitoring (MRM) was used for detection with the following transition 566.6 → 266.4.
LLOQ and ULOQ were at 2 and 2500 ng/ml, respectively. QCs used for extraction recovery and matrix effect was set at 20 and 500 ng/ml at least in triplicate. Extraction recovery for each analyzed compartment is reported in Supplementary Table
4
Quantification of Glipizide in Dissolution Studies
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A reversed phase HPLC method was developed in-house to quantify glipizide in samples obtained from solubility and dissolution studies. The analysis was performed on the Perkin Elmer HPLC system (Series 200) at a temperature of 30 ± 2 °C. The column used (Luna C8, 100 × 4.6 mm, 3 μm) was from Phenomenex®, CA, USA, while the mobile phase was acetonitrile–potassium dihydrogen orthophosphate buffer (pH 4.5; 20 mM) (35:65, v/v). Mobile phase flow rate, detection wavelength and injection volume were 0.8 ml/min, 226 nm and 20 μl respectively. The method was linear (r2 = 0.999) in the concentration range of 0.05–70 μg/ml. The inter-day and intra-day precision was within the acceptable range of less than 2%.
5
Quantification of GnRH [6-D-Phe] in Precipitates
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The GnRH [6-D-Phe] content in the formed precipitate/fibrils was analysed by RP-HPLC after dissolution in 0.1% acetic acid, using a LUNA C8 (4.6 × 250 mm; size = 5 µm; Phenomenex, USA) column, with a C8 pre-column (4 × 3 mm; size = 5 µm) at an HPLC Agilent 1100/1200 series (Agilent Technologies, USA) (mobile phase A (water +1 mL/L Trifluoroacetic acid (TFA) (v/v)) and mobile phase B (800 g Acetonitrile +200 g water +1.2 mL TFA), 1.1 mL/min flow, column temperature 40 °C, and autosampler temperature 2–8 °C. The Retention Time (RT) of GnRH [6-D-Phe] was 8.5 ± 1.5 minutes with UV detection at 220 nm.
6
Chlorophyll and Carotenoid Quantification
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Chlorophyll concentration was determined spectrophotometrically in triplicate samples. 5 mL samples were centrifuged in glass tubes for 8 min at 2650g in an ALC-PK110 centrifuge. The supernatant was discarded, and the pellet was resuspended in 5 mL of pure methanol. The tubes were then placed in a 70 °C water bath for 3 min, and centrifuged again for 8 min at 2650g. The supernatant absorbance was measured at 665 and 750 nm against a pure methanol blind. The concentrations of individual carotenoids were assessed using a reversed-phase Beckman System Gold HPLC (module 125 solvent) equipped with a diode array detector, model 168 Nouveau (Beckman Instruments, Inc., CA, USA), with a column Luna, C8 (Phenomenex), in accordance with Van Heukelem and Thomas [37 (link)].
7
Quantitative Sphingolipid Analysis by LC-MS/MS
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Sphingolipids were analyzed by LC/ESI-MS/MS based on the method described with some modifications as described below [62 (link)]. Lipids were extracted from cell pellets (equivalent to 600 μg to 1 mg of protein depending on the experiment) using an azeotrophic mix of isopropanol:water:ethyl acetate (3:1:6; v:v:v). Internal standards (50 pmol of d17 long-chain bases and C12 acylated sphingolipids) were added to samples at the onset of the extraction procedure. Extracts were separated on an Agilent 1100 HPLC system. Mobile phases were as described [62 (link)], but with 0.2% formic acid on a Phenomenex Luna C8 (3 μm) 2.1 mm ID × 5 cm column maintained at 60°C for separation of the sphingoid bases and 1-phosphates or a Supelco 2.1 mm ID x 5 cm NH2 column for acylated sphingolipids. The eluate was analyzed with an inline ABI 4000 Q Trap (SCIEX, Framingham, MA) mass spectrometer equipped with a turbo ion spray source. Mass spectrometry settings were set to compensate for differences in ionization efficiency of different acyl chain lengths. The peak areas for the different sphingolipid subspecies were compared with that of the internal standards with 13C isotopic signals compensated for where necessary. All data reported are based on monoisotopic mass and are represented as pmol/mg protein.
8
RP-HPLC Purification of Peptides and Glycopeptides
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RP-HPLC analysis and semi-preparative purifications were performed as follows:
Analytical separations were done in an LC-2010A system (Shimadzu, Kyoto, Japan) using Luna C8 (3 µm, 50 mm × 4.6 mm) and Sphereclone C18 (5 µm, 250 mm × 10 mm) columns (Phenomenex, Torrance, CA, USA) for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.036% TFA in ACN) into solvent A (0.045% TFA in H2O) were used over 15 and 20 min, respectively, at a 1 mL/min flow rate.
Semi-preparative purifications were done in a SCL-10A system (Shimadzu) using Phenomenex Luna C8 (10 µm, 250 mm × 10 mm) and SphereClone C18 (10 µm, 250 mm × 10 mm) columns for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.1% TFA in ACN) into A (0.1% TFA in H2O) over 30 and 20 min, respectively, were used, at a 5 mL/min flow rate.
Analytical separations were done in an LC-2010A system (Shimadzu, Kyoto, Japan) using Luna C8 (3 µm, 50 mm × 4.6 mm) and Sphereclone C18 (5 µm, 250 mm × 10 mm) columns (Phenomenex, Torrance, CA, USA) for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.036% TFA in ACN) into solvent A (0.045% TFA in H2O) were used over 15 and 20 min, respectively, at a 1 mL/min flow rate.
Semi-preparative purifications were done in a SCL-10A system (Shimadzu) using Phenomenex Luna C8 (10 µm, 250 mm × 10 mm) and SphereClone C18 (10 µm, 250 mm × 10 mm) columns for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.1% TFA in ACN) into A (0.1% TFA in H2O) over 30 and 20 min, respectively, were used, at a 5 mL/min flow rate.
9
Quantifying SHe Peptide Adsorption on Alum
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SHe peptide and alum samples were gently agitated, and 0.2 ml was transferred to a microfuge tube. The tubes were centrifuged at 5000 x g for 2 minutes to produce a clear supernatant solution. The supernatants were analyzed by RP-HPLC on an Agilent 1100 HPLC system, with a gradient of water/acetonitrile/trifluoroacetic acid, a Phenomenex Luna C8(2), 5 µm, 4.6 × 150 mm column, with UV detection at 215 nm. The solutions were quantified against an external SHe antigen standard of known concentration. The degree of adsorption was calculated as: (Amount Expected-Amount Found) / Amount Expected x 100%.
10
LC-MS/MS Analysis of Organic Compounds
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LC-MS/MS analysis was carried out on a Waters Alliance HPLC with a Waters 996 PDA detector on line with a Q-Tof mass spectrometer (Waters) featured by an ESI source in positive ionization mode. Column: Phenomenex Luna C8 250 × 4.6 mm, 5 μm, 100 A. Eluent A: Water, B: MeOH. Gradient: 90% B to 100% B in 15 mins, holding for 20 mins. Flow: 0.7 ml min−1. PDA: 400–700 nm. Full Mass range: 450–1000 m/z.
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