The UV-vis absorbance spectrum of each drug was first obtained using a UV/vis/nIR spectrophotometer (Jasco V-670, Tokyo, Japan) to select the appropriate absorbance wavelength for HPLC measurements. The concentration of drug in the solution was quantified using HPLC. An aliquot of this sample was mixed with acetonitrile at a ratio of 1:5. Samples were then analyzed with an Agilent 1260 Infinity II HPLC system with an InfinityLab Poroshell 120 EC-C18, 4.6 × 75 mm, 2.7 µm, analytical LC column. The mobile phase was comprised of HPLC plus grade acetonitrile (Sigma Aldrich, MO) and/or double distilled miliQ water, each containing 0.1% trifluoroacetic acid. Chromatographic separation was achieved by gradient elution with acetonitrile over 10 minutes (10% to 95%) at a flow rate of 1 ml/min. For each drug, a standard curve was prepared. A single peak with a specific retention time corresponding to the drug was used for its quantification.
Hplc plus grade acetonitrile
Manufactured by Merck Group
Sourced in Macao, United Kingdom
HPLC plus grade acetonitrile is a high-performance liquid chromatography (HPLC) solvent used as a mobile phase component in analytical and preparative HPLC applications. It is a polar, aprotic solvent that provides high solvating power and low viscosity, enabling efficient separation and detection of a wide range of analytes.
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2 protocols using hplc plus grade acetonitrile
1
Quantitative HPLC Analysis of Drugs
2
HPLC Separation of Neurotransmitters
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HPLC Plus grade acetonitrile (≥99.9%) was purchased from Sigma-Aldrich (Kent, UK). Formic acid (≥99% LC/MS grade, HiPerSolv CHROMANORM ®) was purchased from VWR. Centrifuge tube filter (Corning ® Costar ® Spin-X ®, 0.22 μm Pore CA Membrane, Sterile, 96/Case, Polypropylene) was purchased from Sigma-Aldrich, which was used to filter gut model fluid samples. Analytical standards powder including LC-MS grade dopamine hydrochloride (99%) and L (-)-Epinephrine (99%) were purchased from Alfa Aesar (Lancashire, UK). L-Noradrenaline (98%), Gamma-Aminobutyric acid (99%) and serotonin were purchased from Sigma-Aldrich Co Ltd.
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