Ammonium formate was purchased from Alfa Aesar (Ward Hill, MA). Iodine, N,N-dimethylformamide (DMF), and triethylamine (TEA) were from Jade Scientific (Westland, MI). Internal standard lipids PC(14:0/14:0), PE(14:0/14:0), and PS(14:0/14:0) were purchased from Avanti Polar Lipids (Alabaster, AL). Ammonium bicarbonate, isopropanol, methanol, and water were from J.T. Baker (Phillipsburg, NJ). Chloroform was from EMD Chemicals (Billerica, MA). All solvents used were high performance liquid chromatography grade, and all lipid extraction and storage solvents contained 0.01% butylated hydroxytoluene (BHT) from Sigma Aldrich (St. Louis, MO). 13C1-S,S’-dimethylthiobutanoylhydroxysuccinimide ester (13C1-DMBNHS) was synthesized as previously described [44 (link),46 (link)]. RPMI 1640 cell culture medium and Penicillin-Streptomycin were from GIBCO, Life Technologies (Grand Island, NY). Fetal calf serum (FCS) was from SAFC® Bioscience (St. Louis, MO).
Ps 14 0 14 0
Manufactured by Avanti Polar Lipids
PS-14:0/14:0 is a synthetic phosphatidylserine lipid produced by Avanti Polar Lipids. It has a molecular formula of C28H54NO10P and a molecular weight of 623.73 g/mol. The lipid contains two 14-carbon saturated fatty acid chains.
Automatically generated - may contain errors
Lab products found in correlation
Pe 14 0 14 0, by Avanti Polar Lipids (3 mentions)
Qtrap 5500, by AB Sciex (2 mentions)
Pc 14 0 14 0, by Avanti Polar Lipids (2 mentions)
Pg 14 0 14 0, by Avanti Polar Lipids (2 mentions)
Pa 17 0 17 0, by Avanti Polar Lipids (2 mentions)
Ammonium formate, by Thermo Fisher Scientific (1 mentions)
Butylated hydroxytoluene bht, by Merck Group (1 mentions)
Rpmi 1640 cell culture medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Luna 3u silica column, by Phenomenex (1 mentions)
Kinetex 2.6μ c18 column, by Phenomenex (1 mentions)
D5 tag 48 0, by CDN Isotopes (1 mentions)
4me 16 0 diether dg, by Avanti Polar Lipids (1 mentions)
1260 hplc system, by Agilent Technologies (1 mentions)
3 protocols using ps 14 0 14 0
1
Lipid extraction and quantification protocol
2
Lipidomic Analysis of Insect Fat Body
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Lipids were extracted from wandering 3rd instar larval fat body as previously described [60 (link)]. The lipidomic analyses were carried out on an analytical system comprising an Agilent HPLC 1260 coupled with a SCIEX 5500 QTRAP. Separation of individual classes of polar lipids by normal phase HPLC was carried out using a Phenomenex Luna 3u silica column (i.d. 150x2.0 mm). Multiple reaction monitoring (MRM) transitions were set up for quantitative analysis of various polar lipids. Individual lipid species were quantified by referencing to spiked internal standards. PC-14:0/14:0, LPC-C20, PE-14:0/14:0, PS-14:0/14:0, PA-17:0/17:0, PG-14:0/14:0 were obtained from Avanti Polar Lipids and dioctanoyl phosphatidylinositol (PI, 16:0-PI) was obtained from Echelon Biosciences, Inc. Separation of glycerol lipids (DAG and TAG) by reverse phase HPLC/ESI/MS/MS was carried out on a Phenomenex Kinetex 2.6μ-C18 column (i.d. 4.6x100mm). Using neutral loss-based MS/MS techniques, the levels of TAG were calculated as relative contents to the spiked d5-TAG 48:0 internal standard (CDN Isotopes), while DAG species were quantified using 4ME 16:0 Diether DG as an internal standard (Avanti Polar Lipids). Free cholesterols and ergosterols were analyzed using HPLC/APCI/MS/MS with the corresponding d6-Cho (CDN Isotopes) as the internal standard.
3
HPLC-MS/MS Analysis of Polar Lipids
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Polar lipids were analyzed using an Agilent 1260 HPLC system coupled with a triple quadrupole/ion trap mass spectrometer (5500Qtrap; SCIEX) as described previously39 (link). Separation of individual lipid classes of polar lipids by normal phase (NP)-HPLC was carried out using a Phenomenex Luna 3μ-silica column (internal diameter 150 × 2.0 mm) with the following conditions: mobile phase A (chloroform: methanol: ammonium hydroxide, 89.5:10:0.5) and mobile phase B (chloroform: methanol: ammonium hydroxide: water, 55: 39: 0.5: 5.5). MRM transitions were set up for comparative analysis of various polar lipids. Individual lipid species were quantified by referencing to spiked internal standards. PC-14:0/14:0, PE-14:0/14:0, PS-14:0/14:0, PA-17:0/17:0, PG-14:0/14:0, LPA-17:0 and LPC-17:0 were obtained from Avanti Polar Lipids (Alabaster, AL) and LIPID MAPS.
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