Agilent 1200 series uv detector

The CoQ₁₀ concentration of all neuronal cell samples in this study was determined using reverse-phase HPLC with UV detection at 275 nm, following the method previously described by Duncan, et al. [36 (link)]. A hexane: Ethanol (5:2) solution was used to extract the CoQ₁₀ from the cells, following the method previously described by Duberley, Abramov, Chalasani, Heales, Rahman and Hargreaves [34 (link)]. Once extracted, CoQ₁₀ was resuspended in HPLC grade ethanol, and separation and quantification was achieved using a reverse phase c18 column (Phenomenex, Torrance, CA, USA) using a mobile phase comprised of methanol, ethanol and perchloric acid (700:300:1.2) at a flow rate of 0.7 mL/min. CoQ₁₀ detection was achieved using a Agilent 1200 series UV detector at 275 nm and its concentration was calculated through comparison with standard CoQ₁₀ solution.

All reagents and solvents were used as purchased from commercial sources or indicated otherwise. Flash chromatography was performed using silica gel (300 mesh). All reactions were monitored by TLC, using silica gel plates with the fluorescence F₂₅₄ displayed by UV light visualization. ¹H NMR and ¹³C NMR spectra were recorded on a Bruker AV-400 spectrometer or Bruker AV-300. Coupling constants (J) are expressed in hertz (Hz). Chemical shifts (δ) of NMR are reported in parts per million (ppm) units relative to an internal control (TMS). Low resolution ESI-MS data were recorded on a Finnigan LCQ Advantage MAX mass spectrometer and high resolution ESI-MS data on an Applied Biosystems Q-STAR Elite ESI-LC-MS/MS mass spectrometer. The purity of the compounds was determined by reverse phase high performance liquid chromatography (HPLC) analysis to be >95%. HPLC was performed on either LC-100 liquid chromatograph equipped with a tunable LC-100 UV detector (Shanghai Wufeng Inc., China) or Agilent 1200 series liquid chromatograph equipped with an Agilent 1200 Series UV detector (Agilent Technologies, USA). The columns used were Cosmosil 5C18 (Nacalai Tesque Inc., Japan) for general purification. A flow rate of 1.0 mL/min was used with mobile phase of MeOH in H₂O with 0.1% modifier (ammonia or trifluoroacetate, v/v).