LC-MS grade methanol and acetonitrile as well as adenosine (CAS# 58-61-7), hypoxanthine (CAS# 68-94-0), succinyladenosine (CAS# 4542-23-8), pantothenate (CAS# 79-83-4), propionylcarnitine (CAS# 17298-37-2), sphinganine 1-phosphate (CAS# 19794-97-9), sphingosine 1-phosphate (CAS# 26993-30-6), uridine monophosphate (UMP, CAS# 58-97-9), α-hydroxybutyrate (AHB, CAS# 600-15-7), β-hydroxybutyrate (BHB, CAS# 300-85-6), 2-hydroxy-3-methylbutyrate (CAS# 4026-18-0), decanoylcarnitine (CAS# 1492-27-9), dodecanoylcarnitine (CAS# 25518-54-1), claycomb medium, fetal bovine serum (FBS), penicillin–streptomycin, norepinephrine, ascorbic acid, glutamine, trypsin-EDTA, trypsin inhibitor type I-S, soybean, Dulbecco’s PBS, gelatin, fibronectin, glucose and all salts used in buffer solutions for cell experiments were from Sigma-Aldrich. 13C4 3-Hydroxybutyric acid was purchased by Cambridge Isotope Laboratories (Andover, MA, USA), (R)/(S)-methyl-2-hydroxybutyric acid (MHB) from Small Molecules, inc. (Hoboken, MA, USA), and propidium iodide and hoechst33342 from Thermo Fischer Scientific Inc. (Waltham, MA, USA). MilliQ water was produced freshly using a Direct-Q-3 apparatus (Millipore, Bedford, MA, USA). Finally, HL-1 cardiomyocytes were a kind gift from Dr. Claycomb (Louisiana State University, New Orleans, LA, USA).
Direct q 3 apparatus
Manufactured by Merck Group
Sourced in United States, United Kingdom
The Direct-Q-3 apparatus is a water purification system designed to produce ultrapure water. It utilizes a combination of filtration and deionization techniques to remove impurities and produce high-quality water suitable for various laboratory applications.
Automatically generated - may contain errors
2 protocols using direct q 3 apparatus
1
Metabolite Analysis of Cardiomyocytes
2
Senicapoc Quantification in Biological Samples
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Senicapoc was obtained from Sigma-Aldrich (Schnelldorf, Germany). Diazepam-D5 and 11-hydroxy-Δ⁹-tetrahydrocannabinol-D3 (THC-OH-D3) were purchased from Cerilliant (Round Rock, Texas). Methanol (MeOH), acetonitrile (MeCN), dimethyl sulphoxide (DMSO) and formic acid (FA) were obtained from Sigma-Aldrich. Water was purified using a Direct-Q 3 apparatus (Millipore, Bedford, MA).
Stock solutions of Senicapoc (1 mg/mL) were prepared in DMSO. Separate stock solutions (0.1 or 1 mg/mL) of the deuterated substances were obtained in MeOH. Standard solutions to spike the samples and prepare the calibrants were prepared by diluting the stock solutions with MeOH. An IS solution containing 0.05 μg/mL diazepam-D5 and 0.5 μg/mL THC-OH-D3 was prepared in MeOH. Mobile phases A and B consisted of 0.1 % FA and 0.1 % FA in MeCN, respectively.
Stock solutions of Senicapoc (1 mg/mL) were prepared in DMSO. Separate stock solutions (0.1 or 1 mg/mL) of the deuterated substances were obtained in MeOH. Standard solutions to spike the samples and prepare the calibrants were prepared by diluting the stock solutions with MeOH. An IS solution containing 0.05 μg/mL diazepam-D5 and 0.5 μg/mL THC-OH-D3 was prepared in MeOH. Mobile phases A and B consisted of 0.1 % FA and 0.1 % FA in MeCN, respectively.
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