Concentrations of glucose, succinate, acetate, citrate, malate, lactate and formate were quantified by HPLC (Finnigan Surveyor, Thermo Fisher Scientific, Waltham, USA). Detection was performed by refractive index (RI) (RID Agilent 1200, Agilent Technologies, CA, USA) with a 300 mm × 7.8 mm Aminex HPX-87H ion exchange column (Bio-Rad, Hercules, CA, USA). 20 µL sample was injected into the mobile phase of 5 mM H2SO4 (aq.), temperature was 60 °C and flow rate 0.35 mL min− 1. The results from HPLC analysis were used together with DW measurements to calculate yield coefficients. Triplicate datasets are considered using error bars.
Rid agilent 1200
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1200 RID (Refractive Index Detector) is a high-performance liquid chromatography (HPLC) detector designed to measure the refractive index of liquids. It provides sensitive and accurate detection of a wide range of analytes, making it a valuable tool for applications such as polymer analysis, carbohydrate analysis, and general HPLC.
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2 protocols using rid agilent 1200
1
HPLC Analysis of Metabolites
2
Growth and Metabolite Analysis
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Growth was followed with OD 600 measurements using a spectrophotometer (Shimadzu UV MINI 1240, Japan). Dry cell weight measurements were done with 5 mL of cultivation broth according to 55 . Concentrations of glucose, succinate, acetate, ethanol, glycerol, lactate, formate and pyruvate were determined by HPLC (Agilent 1100, Agilent Technologies, USA) with a 300 mm × 7.8 mm Aminex HPX-87H ion exchange column (Bio-Rad, Hercules, USA), refractive index detector (RID Agilent 1200, Agilent Technologies, USA) and UV detector (Agilent 1100, Agilent Technologies, USA) set to 210 nm. The mobile phase was 5 mM H 2 SO 4 , temperature 60 °C and flow rate 0.6 mL.min -1 . The results from HPLC and dry cell weight measurements were used to calculate yield coefficients. All measurements were performed in triplicates.
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