A high-performance liquid
chromatography (HPLC) system purchased from PerkinElmer was used for
the quantification of the stevioside in various treated SARCs of S. rebaudiana according to the recent protocol of
Ahmad et al.45 (link) The specifications/parameters
of the HPLC system were adjusted so as to quantify the maximum amounts
of stevioside in SARCs. The HPLC was attenuated with the most commonly
used column (C18) with a fixed size (150 × 4.6 mm)
and particle diameter (5 μm), multiple wavelength detector,
vacuum degasser for the solvent, and a quaternary pump with an autosampler
with the maximum injection volume of 10 μL. Two different solutions
(A & B) were used for the mobile phase, such as 25% pure water
(A) and 75% acetonitrile (B), with a constant flow rate of 1.0 mL/min
using a 10 μL injection volume of maximum stevioside quantity.
A standard chromatogram was obtained by running the standard stevioside
(Sigma; Germany) with similar conditions, and the chromatograms of
each treated SARCs was compared with it. Stevioside quantification
was carried out on the basis of the retention of the standard, and
the results of the stevioside content were measured in mg/g-DW, while
stevioside production was expressed in mg/L DW.
chromatography (HPLC) system purchased from PerkinElmer was used for
the quantification of the stevioside in various treated SARCs of S. rebaudiana according to the recent protocol of
Ahmad et al.45 (link) The specifications/parameters
of the HPLC system were adjusted so as to quantify the maximum amounts
of stevioside in SARCs. The HPLC was attenuated with the most commonly
used column (C18) with a fixed size (150 × 4.6 mm)
and particle diameter (5 μm), multiple wavelength detector,
vacuum degasser for the solvent, and a quaternary pump with an autosampler
with the maximum injection volume of 10 μL. Two different solutions
(A & B) were used for the mobile phase, such as 25% pure water
(A) and 75% acetonitrile (B), with a constant flow rate of 1.0 mL/min
using a 10 μL injection volume of maximum stevioside quantity.
A standard chromatogram was obtained by running the standard stevioside
(Sigma; Germany) with similar conditions, and the chromatograms of
each treated SARCs was compared with it. Stevioside quantification
was carried out on the basis of the retention of the standard, and
the results of the stevioside content were measured in mg/g-DW, while
stevioside production was expressed in mg/L DW.
