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5 protocols using hplc grade 1 propanol

1

Lipid Analysis using LC-MS

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LC-MS grade methanol, dichloromethane, and ammonium acetate were purchased from Fisher Scientific (Pittsburgh, PA) and HPLC grade 1-propanol from Sigma-Aldrich (Saint Louis, MO). Milli-Q water was obtained from an in-house Ultrapure Water System by EMD Millipore (Billerica, MA). The Lipidyzer isotope labeled internal standards mixture consisting of 54 isotopes from 13 lipid classes was purchased from Sciex (Framingham, MA).
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2

LC-MS Lipid Quantification Protocol

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Materials: LC–MS grade methanol, dichloromethane, and ammonium acetate were purchased from Fisher Scientific (Pittsburgh, PA) and HPLC grade 1-propanol was purchased from Sigma-Aldrich (Saint Louis, MO). Milli-Q water was obtained from an in-house Ultrapure Water System by EMD Millipore (Billerica, MA). The Lipidyzer isotope labeled internal standards mixture consisting of 54 isotopes from 13 lipid classes was purchased from Sciex (Framingham, MA).
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3

Liquid Chromatography/MS Analysis of Collagen and Elastin Cross-links

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Liquid chromatography/MS–grade methanol was purchased from VWR (Randor, PA, USA). The following reagents were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA): ion chromatography–grade heptafluorobutyric acid (HFBA), analytic-grade 6N hydrochloric acid (HCl), sodium borohydride (NaBH4), sodium hydroxide, HPLC-grade butanol, HPLC-grade 1-propanol, sodium acetate trihydrate, citric acid monohydrate, glacial acetic acid (AcOH), Chloramine-T, dimethylaminobenzaldehyde, ACS-grade perchloric acid, HPLC-grade acetonitrile (ACN), and L-hydroxyproline (OH-pro). Necessary standards for collagen cross-link analysis were obtained from various sources. Dihydroxylysinonorleucine (DHLNL) and LNL were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Pyridinoline (PYD)/deoxypyridinoline (DPYD) HPLC calibrator was purchased from Quidel Corp. (San Diego, CA, USA) and Pent was purchased from Cayman Chemical (Ann Arbor, MI, USA). The elastin cross-links desmosine (DES) and isodesmosine (IDE) were also commercially available from MP Biomedicals (Santa Ana, CA, USA). We also obtained as a kind gift from Simon P. Robins (University of Aberdeen, Aberdeen, Scotland, UK), the major trifunctional corneal cross-link HHL as well as the difunctional cross-link hydroxylysinonorleucine (HLNL).
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4

Lipidyzer Internal Standards LC-MS Workflow

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All LC-MS grade solvents including acetonitrile and water were purchased from Fisher Optima grade, Fisher Scientific (Pittsburgh, PA, USA). High purity formic acid (99%) LC–MS grade methanol, dichloromethane, and ammonium acetate were purchased from Fisher Scientific and HPLC grade 1-propanol, debrisoquine, and 4-nitrobenzoic acid were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Milli-Q water was obtained from an in-house Ultrapure Water System by EMD Millipore (Billerica, MA, USA). The Lipidyzer isotope-labeled internal standards mixture consisting of 54 isotopes from 13 lipid classes was purchased from SCIEX (Framingham, MA, USA).
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5

Lipid Profiling with Isotope-Labeled Standards

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Materials: Materials: LC–MS grade methanol, dichloromethane, and ammonium acetate were purchased from Fisher Scientific (Pittsburgh, PA) and HPLC grade 1-propanol was purchased from Sigma-Aldrich (Saint Louis, MO). Milli-Q water was obtained from an in-house Ultrapure Water System by EMD Millipore (Billerica, MA). The Lipidyzer isotope labeled internal standards mixture consisting of 54 isotopes from 13 lipid classes was purchased from Sciex (Framingham, MA).
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