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2 protocols using l homoarginine hydrochloride

1

Mass Spectrometry Protocol for Metabolite Analysis

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Purified water was obtained using the Milli-Q Integral 3 system (Millipore SAS, Molsheim, France). Zomepirac sodium salt (used as an internal standard, IS), L-serine, L-arginine hydrochloride monohydrate, ascorbic acid, L-homoarginine hydrochloride, LC-MS grade methanol acetonitrile, formic acid and LC grade ethanol were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Pure p. a. ammonium solution (25%) was purchased from Avantor Performance Materials (Gliwice, Poland). The API-TOF reference mass solution kit (G1969–850001) and tuning solutions, ESI-L low concentration tuning mix (G1969–85000) and ESI-TOF Biopolymer Analysis reference masses (G1969–850003) were purchased from Agilent Technologies (Santa Clara, California, United States).
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2

Male Wistar Rat L-homoarginine Study

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The current study was conducted on 33 male Wistar rats at the age of 6-7 weeks (Charles River Laboratories, Sulzfeld, Germany). Male animals were consistently used to prevent distortion of data by hormonal fluctuations, and to ensure comparability to previous investigations. The animals were housed under standardized conditions with water and food ad libitum. L-homoarginine hydrochloride was purchased by Sigma-Aldrich (Sigma-Aldrich Chemie GmbH, Hamburg, Germany). All experiments adhered to the "Guide for the Care and Use of Laboratory Animals" published by the U.S. National Institutes of Health (NIH Publication No. 85-23, revised 1996). The animal protocol was approved by the local authorities of the Regierungspra ¨sidium Karlsruhe (Karlsruhe, Germany), and conformed with the ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments).
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