During the fermentation, culture supernatant was obtained by centrifuging (MiniSpin Eppendorf, Hamburg, Germany) fermentation broth samples in Eppendorf tubes at 12,000 g for 3 min. The glucose concentration in supernatant was measured with a YSI 2700 glucose analyzer (Yellow Springs Instruments, Yellow Springs, OH, USA). Glucose and organic acid concentrations of filtered supernatants (Minisart filters 0.2 µm size pore, Sartorius, Göttingen, Germany) were determined by HPLC (Ultimate 3000, Dionex, Thermo Fisher Scientific, Courtaboeuf, France) using an Aminex HPX-87H+ column (Bio-Rad Life Science, Marnes-la-Coquette, France) under the following conditions: 50 °C, 5 mM H2SO4 as mobile phase at a flow rate of 0.5 mL·min−1 and detection with a refractometer and an UV detector at 210 nm. The samples were previously diluted in deionised water. External standards (0.2 to 5 g·L−1) were used for compound identification and quantification. Chromatograms were analyzed with Chromoleon software 6.80 (Thermo Fisher Scientific, Courtaboeuf, France). For the flask experiments, the glucose uptake and the biomass yield were calculated at the end of the growth phase.
Ysi 2700 glucose analyzer
Manufactured by Yellow Springs Instruments
Sourced in United States
The YSI 2700 glucose analyzer is a laboratory instrument designed to measure the concentration of glucose in a sample. It provides accurate and reliable glucose readings using electrochemical detection technology. The YSI 2700 is a self-contained unit that requires minimal sample preparation and delivers results quickly.
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2 protocols using ysi 2700 glucose analyzer
1
Fermentation Glucose Quantification by HPLC
2
Oral Glucose Tolerance Test Protocol
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A standard OGTT was performed before and following 16 weeks of ivacaftor treatment. A 50% glucose solution at a dose of 1.75 g/kg body weight (maximum 75 g) was administered orally. Plasma glucose levels were assessed in venous blood samples at baseline and at 30, 60, 90 and 120 min using a YSI 2700 glucose analyzer (Yellow Springs Instruments, Yellow Springs, OH). Concomitant insulin levels were measured by radioimmunoassay (Linco Research, St. Charles, MO). The area under the curve (AUC) for glucose and insulin was calculated using the trapezoidal estimation. The study was approved by the Institutional Review Board of the Hadassah-Hebrew University Medical Center and written informed consent was obtained from the study participants.
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