Liquid chromatography - mass spectrometry (LC–MS) grade methanol and high-performance liquid chromatography (HPLC) grade isopropanol were purchased from Millipore Sigma (Billerica, MA). Formic acid, ammonium formate, and ammonium hydroxide were purchased from Sigma-Aldrich (St. Louis, MO). HPLC grade chloroform was purchased from Honeywell Burdick & Jackson (Muskegon, MI). Glass beads (0.5 mm) were purchased from Bertin Technologies (Montigny, France). Stock solutions of lipid standards, purchased from Avanti (Alabaster, AL), were prepared in chloroform and kept at − 80 °C until analysis. The columns for LC–MS analysis were purchased from Phenomenex (Torrance, CA) and included a C5 Luna column (50 × 4.6 mm, 5 μm) and a C18 Gemini column (50 × 4.6 mm, 5 μm) for electrospray ionization positive (ESI+) and negative (ESI−) modes. Nanopure water was used for all analyses (resistivity 18.2 MΩ-cm at 25 °C).
Chloroform hplc grade
Manufactured by Avantor
Sourced in United States
Chloroform (HPLC grade) is a colorless, volatile liquid commonly used as a solvent in high-performance liquid chromatography (HPLC) applications. It has a high density and a distinctive, pleasant odor. This product meets the specifications required for HPLC-grade solvents, ensuring consistent performance and reliable results in analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
Ammonium formate, by Merck Group (2 mentions)
Methanol, by Avantor (2 mentions)
Glass beads, by Bertin Technologies (1 mentions)
Gemini c18 column, by Phenomenex (1 mentions)
Ammonium hydroxide, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Toluene, by Avantor (1 mentions)
Protein assay dye reagent, by Bio-Rad (1 mentions)
Hplc grade methanol, by Avantor (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
4000 qtrap, by AB Sciex (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
Lc ms grade methanol, by Scharlab (1 mentions)
Ms grade ammonium formate, by Merck Group (1 mentions)
Ultrapure water, by Thermo Fisher Scientific (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
9 protocols using chloroform hplc grade
1
LC-MS Lipid Analysis Protocol
2
Organic Solvent Extraction and Quantification
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HPLC grade chloroform (CAS: 67-66-3), dichloromethane (CAS: 75-09-2), and toluene (CAS: 108-88-3) were purchased from Honeywell Burdick & Jackson (Muskegon, MI, USA); protein assay dye reagent and acetylacetone were purchased from Bio-Rad (CA, USA). All other reagents were purchased from Sigma-Aldrich.
3
Lipidomic Analysis of Cell Cultures
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DMEM, RPMI, FBS, and penicillin/streptomycin (100 U/ml penicillin G sodium and 100 μg/ml streptomycin sulfate) were obtained from Invitrogen Life Technologies (Carlsbad, CA). The HPLC used was a Shimadzu Prominence LC-20-AD system, and the mass spectrometer was a 4000 QTRAP® from ABSciex. Prior to mass spectrometric analysis, lipids were separated by reverse-phase chromatography using a Phenomenex Kinetex 2.6 μm C18 100A 50 × 2.1 mm reverse-phase HPLC column (Torrance, CA). HPLC-grade methanol, HPLC-grade chloroform, and American Chemical Society-grade formic acid (EMD Chemicals) were purchased from VWR (Bridgeport, NJ).
4
Preparation of High-Purity Organic Solvents
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Ultrapure CO2 (99.999%) was provided by Air Products (Amsterdam, Netherlands). LC–MS-grade methanol was purchased from Scharlau (Barcelona, Spain). Heptane (99.5%) was purchased from Alfa Aesar (Karlsruhe, Germany). Ethanol 99.7% and HPLC-grade chloroform were from VWR (VWR Chemicals, France). MS-grade ammonium formate was purchased from Sigma-Aldrich (St. Louis, MO). Water was purified using a Milli-Q purification system (Millipore, Billerica, MA).
5
Lipid Extraction and Characterization
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HPLC‐grade chloroform, MeOH, and water used for sample pretreatment were supplied by VWR International (Milan, Italy). Ultrapure water and isopropanol (i‐PrOH) of LC‐MS grade were purchased from Thermo Fisher Scientific (Waltham, MA, USA); LC‐MS grade MeOH was provided by Romil Pure Chemistry (Pozzuoli, NA, Italy).
Ammonium acetate, acetic acid, sulfuric acid, and n‐hexane were purchased from Merck (St. Louis, MO, USA). The 35 lipid standards constituted of 28 FAs, 1 TAG, 1 diacylglycerol (DAG), and 5 phospholipids are reported in Supporting information TableS1 with all valuable information.
Ammonium acetate, acetic acid, sulfuric acid, and n‐hexane were purchased from Merck (St. Louis, MO, USA). The 35 lipid standards constituted of 28 FAs, 1 TAG, 1 diacylglycerol (DAG), and 5 phospholipids are reported in Supporting information Table
6
Metabolomics and Spatial Lipidomics Protocols
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For metabolomics experiments, LC-MS grade methanol, LC-MS grade water, LC-MS grade acetonitrile, LC-MS grade 2-propanol, formic acid (99.5+%), ammonium acetate, ammonium formate and ammonium hydroxide were purchased from Fisher Chemical (Fisher Scientific International, Inc., Pittsburgh, PA, USA) and used to prepare chromatographic mobile phases. Isotopically labeled lipid standards (Table S1a ) were purchased from Avanti Polar Lipids (Alabaster, AL, USA) and used to prepare the lipid internal standard mixture. Isotopically labeled polar metabolite standards (Table S1b ) were purchased from Cambridge Isotope Laboratories (Tewksbury, MA, USA) and were used to prepare the internal standard mixture for HILIC experiments.
For spatial lipidomics experiments, carboxymethyl cellulose (CMC), gelatin from bovine skin (Type B), 1,5-naphthalenediamine (1,5-DAN, ≥97%) and isopentane (≥95%) were purchased from Sigma Aldrich (St. Louis, MO, USA) and used as received. Acetonitrile (HPLC grade), methanol (HPLC grade) and chloroform (HPLC grade) were purchased from VWR and also used as received.
For spatial lipidomics experiments, carboxymethyl cellulose (CMC), gelatin from bovine skin (Type B), 1,5-naphthalenediamine (1,5-DAN, ≥97%) and isopentane (≥95%) were purchased from Sigma Aldrich (St. Louis, MO, USA) and used as received. Acetonitrile (HPLC grade), methanol (HPLC grade) and chloroform (HPLC grade) were purchased from VWR and also used as received.
7
Quantifying Short-Chain Fatty Acids
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Isobutanol HPLC grade (99% purity) was purchased from Alfa Aesar (Haverhill, MA, USA). The three internal standards (IS) were propionic acid d5 (96% purity), butyric acid d8 (96% purity), and 3-methyl pentanoic acid (>98% purity), and were all purchased from Sigma Aldrich (St. Louis, MO, USA). Additionally, pyridine anhydrous (99.8% purity), isobutyl chloroformate, chloroform HPLC grade (99.8% purity), cyclohexane HPLC grade, and the boiling chip granules of 2–8 mm were purchased from VWR chemicals (Radnor, PA, USA); 99.99% pure sodium hydroxide pellets were purchased from Merck (Burlington, MA, USA). The stock solution of SCFAs at 10 mM was purchased from Sigma Aldrich (St Louis, MO, USA). Mili Q water was produced using a Millipore Milli-Q system with LC-PAK Polisher/Millipak Express 40 and was purchased from Merck Millipore (Burlington, MA, USA).
8
Lipidomic Internal Standards Protocol
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1-Heptadecanoyl-2-hydroxy-sn-glycero-3-phosphate (sodium salt) (LPA 17:0), 1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt) (PA 14:0/14:0), N-lauroyl-d-erythrosphingosylphosphorylcholine (SM d18:1/12:0), 1-myristoyl-2-hydroxy-sn-glycero-3-phospho-(1 -rac-glycerol) (sodium salt) (LPG 14:0), 1,2-diheptadecanoyl-sn-glycero-3-phospho-(1 -rac-glycerol) (sodium salt) (PG 17:0/17:0), 1-nonadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC 19:0), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (PC 14:0/14:0), 1,2-dimyristoyl-sn-glycero-3-phospho-l-serine (sodium salt) (PS 14:0/14:0), 1-myristoyl-2-hydroxysn-glycero-3-phosphoethanolamine (LPE 14:0), and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (PE 14:0/14:0) were purchased from Avanti Polar Lipids (Alabaster, USA) and used as internal standards (IS). Water, methanol, acetonitrile, ammonium formate (all LC-MS grade), n-hexane, and chloroform (HPLC grade) were purchased from VWR (Radnor, USA). LC-MS-grade 2-propanol was purchased from Fluka (Buchs, Switzerland) and acetone (GC-MS grade) was from MERCK (Darmstadt, Germany).
9
Biofluids Analytical Standards Protocol
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Methanol, water, isopropyl alcohol (IPA) acetonitrile and chloroform (HPLC grade) were purchased from VWR International (Loughborough, UK). Formic acid (HPLC grade) and ammonium formate (99%+ purity) were purchased from Sigma-Aldrich (Poole, UK). 5mL volumes of human urine and plasma from one male donor were purchased from BioIVT (Sussex, UK). These were delivered frozen, thawed for aliquoting, and were then frozen again at -80 o C before being thawed for analysis, giving a total of two freeze/thaw cycles. 2mL porcine urine samples, 1mL porcine plasma samples and ~20µL porcine basal tear uid samples were all collected across 13 hourly time points by the Defence Science and Technology Laboratory (Porton Down, UK). These porcine samples were obtained from non-recovery anaesthetised animals within a pre-existing programme of animal work carried out by Dstl in accordance with the UK Animals (Scienti c Procedures) Act, 1986. Samples were delivered frozen and only thawed for analysis, giving a total of 1 freeze/thaw cycle.
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