HPLC Analysis of Compounds

HPLC analysis was performed as previously described (12 (link)). Specifically, analysis was performed using 95% water–5% acetonitrile–0.1% trifluoroacetic acid (TFA) as the mobile phase with a flow rate of 0.5 ml/min on a Microsorb-MV C₁₈ column. After a 10-min equilibration period, a linear 40-min acetonitrile gradient (5% to 80%) was employed and the final concentration was maintained for an additional 20 min. The eluate was monitored using a photodiode array detector from 200 to 900 nm by reverse-phase chromatography on a Varian ProStar HPLC instrument.

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Lojek L.J., Farrand A.J., Wisecaver J.H., Blaby-Haas C.E., Michel B.W., Merchant S.S., Rokas A, & Skaar E.P. (2017). Chlamydomonas reinhardtii LFO1 Is an IsdG Family Heme Oxygenase. mSphere, 2(4), e00176-17.

Publication 2017

ProStar HPLC instrument

Acetonitrile Hplc Reverse phase chromatography Trifluoroacetic acid

Corresponding Organization :

Other organizations : Vanderbilt University Medical Center, Vanderbilt University, University of California, Los Angeles, University of Denver

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Variable analysis

independent variables

Acetonitrile gradient (5% to 80%)

dependent variables

Eluate monitored using a photodiode array detector from 200 to 900 nm

control variables

Mobile phase: 95% water–5% acetonitrile–0.1% trifluoroacetic acid (TFA)
Flow rate: 0.5 ml/min
Column: Microsorb-MV C18
Equilibration period: 10 min
Gradient duration: 40 min
Final concentration maintained for 20 min
Reverse-phase chromatography on a Varian ProStar HPLC instrument

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