The hydrophobic interaction chromatography magnetic beads (MB-HIC8), peptide calibration standard and α-cyano-hydroxycinnamic (HCCA) were purchased from Bruker Daltonics. We used acetonitrile (ACN) HPLC grade (Scharlau, Barcelona, Spain) and trifluoroacetic acid (TFA) HPLC grade (Loba Chemie, Mumbai, India). Elution buffer was freshly prepared daily by mixing 50% of ACN with MB-HIC stabilization buffer. HCCA matrix was prepared by dissolving 1.2 mg HCCA in 200 µl ACN, 40 µl 10% TFA and 160 µl distilled water, this yielded 3mg/ml HCCA in 50% ACN: 2% TFA.
Acetonitrile acn hplc grade
Manufactured by Scharlab
Sourced in Spain
Acetonitrile (ACN) HPLC grade is a common organic solvent used in high-performance liquid chromatography (HPLC) applications. It is a clear, colorless liquid with a characteristic odor. Acetonitrile HPLC grade is specifically formulated to meet the purity requirements for HPLC mobile phase preparation and sample preparation.
Automatically generated - may contain errors
Lab products found in correlation
Triisopropylsilane, by Thermo Fisher Scientific (1 mentions)
O 7 azabenzotriazol 1 yl n n n n tetramethyluronium hexafluorophosphate hatu, by GL Biochem (1 mentions)
Fmoc protected amino acids, by GL Biochem (1 mentions)
Milli q system, by Merck Group (1 mentions)
T2modx, by Avantor (1 mentions)
Agilent 1200 hplc, by Agilent Technologies (1 mentions)
Fecl3, by Reanal (1 mentions)
3 protocols using acetonitrile acn hplc grade
1
Hydrophobic Interaction Chromatography Beads Protocol
2
Peptide Synthesis and Characterization
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Solvents for RP-HPLC were acquired as HPLC grade and used without further purification. All other reagents were used as supplied. Conventional Fmoc protected amino acids, O-(6-chlorobenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HCTU) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) were purchased from GL Biochem (Shanghai, China). N-Methylmorpholine (NMM), N,N′-diisopropylcarbodiimide (DIC), 6-chloro-1-hydroxybenzotriazole (6-Cl-HOBt), piperidine and Tris free base were purchased from Aldrich (St Louis, USA). Dichloromethane (DCM), and sodium chloride (NaCl) were purchased from ECP limited (Auckland, New Zealand). Hydrochloric acid (HCl) and sodium hydroxide (NaOH), dimethylformamide (DMF, AR grade) and acetonitrile (ACN, HPLC grade) were purchased from Scharlau (Barcelona, Spain). Trifluoroacetic acid (TFA) was purchased from Oakwood Chemical (River Edge, USA). Triisopropylsilane (TIPS) was purchased from Alfa Aesar (Wardhill, MA). The aminomethyl polystyrene resin was purchased from Rapp Polymer GmbH (Tübingen, Germany). Bovine lactoferrin (LF) was kindly donated by Fonterra Co-operative Group (New Zealand).
TEM images were analyzed with ImageJ 1.50i.33 (link)
TEM images were analyzed with ImageJ 1.50i.33 (link)
3
Quantification of Coenzyme Q10 by HPLC-UV
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Acetonitrile (ACN; HPLC grade) was purchased from Scharlau (Barcelona, Spain). Tetrahydrofuran (THF; isocratic HPLC grade), 0.45 µm Polytetrafluoroethylene PTFE syringe filters were ordered from VWR (Radnor, PA, USA). The standard coenzyme Q10 (≥98%) was obtained from the Sigma-Aldrich group (Schnelldorf, Germany). FeCl 3 (≥99%) were ordered from Reanal (Budapest, Hungary). Ultra-pure water (18.2 Mcm) was obtained from a Milli-Q system from Merck-Millipore (Milford, MA, USA).
For the determination of total Q10 content an Agilent 1200 HPLC (Agilent Technologies) system was used and the chromatography was executed in isocratic mode on an Agilent Zorbax XDB C 18 HPLC column (2.1 mm × 50 mm × 3.5 μm) followed by UV detection at 275 nm. The column temperature was set to 30 °C. The eluent consisted of the mixture of ACN: THF:water in 65:30:5 % v/v rate and the flow rate was 0.35 ml min -1 . The injection volume was 10 µl. The sample preparation was assisted with an ultrasonic bath (type T2MODX; VWR), and with a horizontal shaking table (T2125; MTA Kutesz, Budapest, Hungary) operated at 400 RPM. The limit of quantitation (LOQ) for HPLC-UV method was 0.05 mg Q10/g.
For the determination of total Q10 content an Agilent 1200 HPLC (Agilent Technologies) system was used and the chromatography was executed in isocratic mode on an Agilent Zorbax XDB C 18 HPLC column (2.1 mm × 50 mm × 3.5 μm) followed by UV detection at 275 nm. The column temperature was set to 30 °C. The eluent consisted of the mixture of ACN: THF:water in 65:30:5 % v/v rate and the flow rate was 0.35 ml min -1 . The injection volume was 10 µl. The sample preparation was assisted with an ultrasonic bath (type T2MODX; VWR), and with a horizontal shaking table (T2125; MTA Kutesz, Budapest, Hungary) operated at 400 RPM. The limit of quantitation (LOQ) for HPLC-UV method was 0.05 mg Q10/g.
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