Lysophosphatidylcholine

Manufactured by Avanti Polar Lipids

Sourced in United States

Lysophosphatidylcholine is a lipid compound that is a by-product of the enzymatic cleavage of phosphatidylcholine. It is commonly used as a component in various cell culture media and as a tool in biochemical research.

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26 protocols using lysophosphatidylcholine

Lipidomic Analysis Solvent Standards

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Spectral grade solvents for lipid extraction and LC/MS analysis were obtained from Sigma-Aldrich (St. Louis, MO). The authentic internal standards (IS) for lipidomic analysis were purchased from Sigma-Aldrich and Avanti Polar Lipids (Alabaster, AL), including triglyceride (TG) 11:0/11:0/11:0, phosphatidylcholine (PC)
13:0/13:0, lysophosphatidylcholine (LPC) 15:0, phosphatidylethanolamine (PE) 15:0/15:0, lysophosphatidylethanolamine (LPE) 13:0, phosphatidylinositol (PI) 8:0/8:0, lysophosphatidylinositol (LPI) 13:0, and free fatty acid (FFA) 17:0. While cardiolipin (CL) (15:0)4 was synthesized according to Ahmad et al. (2005) . The mixed solution of all the IS was newly prepared with methanol (containing 0.006% BHT, w/v) and stored at -80 °C until use. Other chemicals and reagents were of analytical grade unless specified.

Chen Z., Liang Q., Wu Y., Gao Z., Kobayashi S., Patel J., Li C., Cai F., Zhang Y., Liang C., Chiba H, & Hui S.P. (2020). Comprehensive lipidomic profiling in serum and multiple tissues from a mouse model of diabetes. Metabolomics : Official journal of the Metabolomic Society, 16(11).

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Lipidomic Analysis: Standards and Solvents

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LC-MS grade acetonitrile, methanol, water, isopropyl alcohol, and HPLC grade methanol, and chloroform were purchased from Thermo Fisher Scientific (Fairlawn, NJ, USA). LC/MS grade eluent buffers, ammonium acetate, and acetic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Diacylglycerol (DG, 10:0/10:0), triacylglycerol (TG, 17:0/17:0/17:0), phosphatidylcholine (PC, 10:0/10:0), phosphatidylethanolamine (PE, 10:0/10:0), sphingomyelin (SM, 18:1d/17:0), and ceramide (CER, C17) for internal standards were acquired from Avanti Polar Lipids (Alabaster, AL, USA). Diacylglycerol (DG, 16:0/16:0), phosphatidylcholine (PC, 16:0/16:0), phosphatidylethanolamine (PE, 16:0/16:0), lyso-phosphatidylcholine (lyso-PC, 18:0), lyso-phosphatidylethanolamine (lyso-PE 16:0), triacylglycerol (TG, 17:0/17:0/17:0), sphingomyelin (SM, 18:1d/17:0), and ceramide (CER, C17) as standards for retention time were obtained from Avanti Polar Lipids (Alabaster, AL, USA).

Hong S.H., Lee J.Y., Seo S., Shin B., Jeong C.H., Bae E., Kim J., Lee D., An B., Shim M., Shin J.H., Lee D.K., Kim Y.J, & Han S.B. (2023). Lipidomic Analysis of Cervicovaginal Fluid for Elucidating Prognostic Biomarkers and Relevant Phospholipid and Sphingolipid Pathways in Preterm Birth. Metabolites, 13(2), 177.

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Measuring LysoPLD Activity in Serum and Protein

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LysoPLD activity was measured as previously described.⁸⁰ (link) Briefly, 10 μL mouse serum or rmATX (2 ug/mL in phosphate-buffered saline) was incubated with 2 mmol/L 1-myristoyl (14:0)– lysophosphatidylcholine (Avanti Polar Lipids, Inc, Alabaster, AL) in 90 μL assay buffer (100 mmol/L Tris–HCl, pH 9.0, 500 mmol/L NaCl, 5 mmol/L MgCl₂, 5 mmol/L CaCl₂, and 0.05% Triton X-100) for 4 hours at 37°C to allow for autotaxin-mediated choline release. Subsequently, 100 μL reaction buffer containing 4.5 mmol/L 4-aminoantioyrine, 2.7 mmol/L TOOS reagent (N-ethyl-N-[2-hydroxy-3-sulfopropyl]-3-methylaniline, a hydrogen donor), 20 U/mL horseradish peroxidase, 3 U/mL choline oxidase, 4.5 mmol/L MgCl₂, 100 mmol/L Tris, pH 8.0) was added and incubated for another 10 minutes at room temperature to allow for color reaction. The absorbance was measured at 555 nm for the calculation of choline production (nmol) per microliter of serum per hour (for serum), or choline production (μmol) per milligram of protein per minute (for rmATX).

Qiu H., Song E., Hu Y., Li T., Ku K.C., Wang C., Cheung B.M., Cheong L.Y., Wang Q., Wu X., Hoo R.L., Wang Y, & Xu A. (2022). Hepatocyte-Secreted Autotaxin Exacerbates Nonalcoholic Fatty Liver Disease Through Autocrine Inhibition of the PPARα/FGF21 Axis. Cellular and Molecular Gastroenterology and Hepatology, 14(5), 1003-1023.

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Lipid Standards for Mass Spectrometry

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Acetonitrile, methanol, and 2-propanol (Optima LC-MS grade) were purchased from Fisher Chemical(Waltham, USA); methyl-tert-butyl ether (MTBE), ammonium formate, ammonium hydrogen carbonate (BioUltra grade) and chloroform (analytical reagent grade) were purchased from Sigma-Aldrich (St. Louis, USA) Deionized water was obtained from an in-house water purification system. The following standards, lyso phosphatidylethanolamine (LPE) 14:0, Lyso phosphatidylcholine (LPC) 13:0, phosphatidylcholine (PC) 13:0_13:0, phosphatidylethanolamine (PE) 14:0_14:0, phosphatidylserine (PS) 14:0_14:0, phosphatidylinositol (PI) 12:0–13:0, ceramide d18:1/17:0, glucosyl(ß) C8 ceramide, hexosyl1ceramide d18:1/8:0 and 06:0 sphingomyelin (SM) were purchased from Avanti Polar Lipids (Alabaster, USA). GM3 d18:1/18:0 d3 was purchased from Matreya LLC (State College, USA). Diacylglycerol (DG) 15:0_15:0 was purchased from Santa Cruz Biotechnology, Inc. (Dallas, USA). Glyceryl triheptadecanoate (TAG) 17:0_17:0_17:0 was purchased from Sigma Aldrich (St. Louis, USA) and glyceryl trihexadecanoate (TAG) 48:0 d5 was purchased from CDN Isotopes (Pointe Claire, Canada)

Selvalatchmanan J., Rukmini A.V., Ji S., Triebl A., Gao L., Bendt A.K., Wenk M.R., Gooley J.J, & Torta F. (2021). Variability of Lipids in Human Milk. Metabolites, 11(2), 104.

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Plasma and Lymph Lipidomic Analysis

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To prepare blood plasma and lymph fluid for lipidomic analysis samples were transferred to glass vials for Bligh-Dyer lipid extraction using methanol:PBS:chloroform (1:1:2) containing 0.1% formic acid. Ten μl of a mix of internal standards were added to each extraction mixture. The internal standards included known quantities of 13:0 lyso-phosphatidyl choline, 12:0–13:0 phosphatidyl choline, and 12:0–13:0 phosphatidyl inositol, purchased as solutions from Avanti Polar Lipids (Alabaster, AL). After vortexing for 1 minute, the samples were centrifuged at 1500xg for 15 minutes. Using a glass Hamilton syringe, the bottom (chloroform) layer of solvent was removed and transferred to a clean screw-cap glass vial. Lipid extracts were concentrated by drying under a compressed air stream (to reduce lipid oxidation) and then reconstituting in 100 μl of chloroform prior to liquid chromatography and mass spectrophotometry.

Ubellacker J.M., Tasdogan A., Ramesh V., Shen B., Mitchell E.C., Martin-Sandoval M.S., Gu Z., McCormick M.L., Durham A.B., Spitz D.R., Zhao Z., Mathews T.P, & Morrison S.J. (2020). Lymph protects metastasizing melanoma cells from ferroptosis. Nature, 585(7823), 113-118.

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Plasma and Lymph Lipidomic Analysis

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Lipid Profiling Internal Standards

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For the authentic internal standards (IS), triacylglycerol (TG) 11:0/11:0/11:0 and free fatty acid (FFA) 17:0 were obtained from Sigma-Aldrich (St. Louis, MO, USA), and phosphatidylcholine (PC) 13:0/13:0, phosphatidylethanolamine (PE) 15:0/15:0, phosphatidylinositol (PI) 8:0/8:0, lysophosphatidylcholine (LPC) 15:0, lysophosphatidylethanolamine (LPE) 13:0, and lysophosphatidylinositol (LPI) 13:0 were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Cardiolipin (CL) 15:0/15:0/15:0/15:0 was synthesized in our laboratory previously [16 (link)]. Other chemicals and reagents, which were of the highest grade available, were purchased from Sigma-Aldrich unless specified. The mixed solution of all the IS was prepared with methanol (containing 0.006% butylated hydroxytoluene, w/v) and stored at −80 °C for no more than two weeks before use.

Wu Y., Chen Z., Fuda H., Tsukui T., Wu X., Shen N., Saito N., Chiba H, & Hui S.P. (2021). Oxidative Stress Linked Organ Lipid Hydroperoxidation and Dysregulation in Mouse Model of Nonalcoholic Steatohepatitis: Revealed by Lipidomic Profiling of Liver and Kidney. Antioxidants, 10(10), 1602.

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Comprehensive Lipidomics and Metabolomics Analysis

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All reagents were LC-MS grade (Fisher Scientific, Hanover Park, IL). Standards for lipidomics included phosphatidylethanolamine (PE 14:0/14:0), phosphatidylcholine (PC 14:0/14:0), lysophosphatidylcholine (LysoPC 17:1), cholesterol, cholesteryl ester (ChoE 19:0) (Avanti Polar Lipids, Inc., Alabaster, AL), TG (19:1/19:1/19:1), diacylglyceride (DG 20:1/20:1), monoacylglyceride (MG 17:1), and free fatty acid (FFA 17:1) (Nu-check Prep Inc., Elysian, MN). Metabolomic standards included debrisoquine sulfate, 4-nitrobenzoic acid, valine, hypoxanthine, carnitine, proline, tyrosine, glucose, uridine, and taurine (all Sigma-Aldrich, St. Louis, MO). All standards were of the highest purity available.

Pannkuk E.L., Laiakis E.C., Mak T.D., Astarita G., Authier S., Wong K, & Fornace AJ J.r. (2016). A Lipidomic and Metabolomic Serum Signature from Nonhuman Primates Exposed to Ionizing Radiation. Metabolomics : Official journal of the Metabolomic Society, 12(5), 80.

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Lipid Standards for Analytical Protocols

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HPLC-grade
acetonitrile (ACN),
methanol (MeOH), and isopropanol (IPA) were purchased from Merck (Darmstadt,
Germany). HPLC-grade dichloromethane (CH₂Cl₂), tert-butyl methyl ether (MTBE), and ammonium
acetate (AmAc) were purchased from Sigma–Aldrich (St. Louis,
MO. USA). Ultrapure water was obtained by a Milli-Q system (Millipore,
Billerica, MA). Lipid standards including phosphatidylcholine (PC)
19:0/19:0, lyso-phosphatidylcholine (LPC) 19:0, phosphatidylethanolamine
(PE) 17:0/17:0, sphingomyelin (SM) d18:1/12:0, ceramide (Cer) d18:1/17:0,
triacylglycerol (TG) 15:0/15:0/15:0, and fatty acid (FA) 16:0-d3 were
purchased from Avanti Polar Lipids (Alabaster, AL).

Xuan Q., Hu C., Yu D., Wang L., Zhou Y., Zhao X., Li Q., Hou X, & Xu G. (2018). Development of a High Coverage Pseudotargeted Lipidomics Method Based on Ultra-High Performance Liquid Chromatography–Mass Spectrometry. Analytical Chemistry, 90(12), 7608-7616.

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Comprehensive Lipidomics Analytical Standards

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HPLC-grade acetonitrile (ACN), methanol (MeOH), and isopropanol (IPA) were purchased from Thermo Fisher Scientific, U.S.A. HPLC-grade methyl-tert-butyl ether (MTBE), ammonium formate, and formic acid were purchased from Sigma–Aldrich (St. Louis, MO, U.S.A.). Ultrapure water was obtained by a Milli-Q system (Millipore, Billerica, MA). SPLASH internal standards (330707, SPLASH™ Lipidomix Mass Spec Standards) were purchased from Avanti Polar Lipids (Alabaster, U.S.A.), including lyso-phosphatidylcholine (LPC) 18:1 (d7), 25 μg/ml; lyso-phosphatidylethanolamine (LPE) 18:1 (d7), 5 μg/ml; phosphatidylcholine (PC) 15:0-18:1 (d7), 160 μg/ml; phosphatidylethanolamine (PE) 15:0-18:1 (d7), 5 μg/ml; phosphatidylglycerol (PG) 15:0-18:1 (d7), 30 μg/ml; phosphatidylserine (PS) 15:0-18:1(d7), 5 μg/ml; phosphatidylinositol (PI) 15:0-18:1 (d7), 10 μg/ml; phosphatidic acid (PA) 15:0-18:1 (d7), 7 μg/ml; sphingomyelin (SM) d18:1-18:1 (d9), 30 μg/ml; cholesterol (d7), 100 μg/ml; ceramide (Cer) 18:1 (d7), 350 μg/ml; monoglyceride (MG) 18:1 (d7), 2 μg/ml; diglyceride (DG) 15:0-18:1 (d7), 10 μg/ml; and triglyceride (TG) 15:0-18:1 (d7)-15:0, 55 μg/ml

Bai Y., Huang W., Li Y., Lai C., Huang S., Wang G., He Y., Hu L, & Chen C. (2021). Lipidomic alteration of plasma in cured COVID-19 patients using ultra high-performance liquid chromatography with high-resolution mass spectrometry. Bioscience Reports, 41(3), BSR20204305.

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