HPLC analysis for withanolides was performed with some modification to the method of Mundkinajeddu et al. [25 ]. The Jasco HPLC system (LC-Net II), consisting of a quaternary pump (PU-2089), an autosampler (AS-2059 Plus) and a PDA detector (MDA-2018) was used for putative detection of withanolides. The compounds separation were achieved using 5 μm Luna C18 250 × 4.6 column and Solvents (A = Water Ultra-Pure + 0.05% TFA (pH. 2.6), B = MeOH HPLC grade). Mobile phase was run using gradient elution at 0, 15, 20 and 23 min of 30, 80, 80 and 30% of solvent-B respectively. The flow rate was 0.7 ml/min and injection volume was 20 μl. The eluent was detected and analysed at 230 nm. To get a clearer picture of the variation in root samples we integrated the peaks area region of 15–26 min which were assigned as global withanolides area (data not shown). For background correction standards of chlorogenic acid (100 mg L−1), and withaferin A (146 μM) were passed through HPLC to assign respective peaks.
As 2059 plus
Manufactured by Jasco
Sourced in Japan
The AS-2059 Plus is a laboratory equipment designed for general use. It serves as a basic tool for various scientific applications. The core function of this product is to provide a reliable and efficient platform for conducting various experiments and analyses in a laboratory setting.
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2 protocols using as 2059 plus
1
HPLC Analysis of Withanolides in Herbal Extracts
2
Polyphenols Analysis by Integrated HPLC
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Polyphenols analysis was performed by LC-4000 Series Integrated HPLC Systems (JASCO, Tokyo, Japan) consisting of a column oven (model CO-2060 plus), set at 30 °C, a UV/Vis photodiode array detector (model MD-2018 plus), an intelligent fluorescence detector (model PF-2020 plus), a liquid chromatography pump (model PU-2089 plus), an autosampler (AS-2059plus) and a ChromNAV software program (JASCO, Japan). A C18 Luna column 5 μm particle size, 25 cm × 3.00 mm I.D. (Phenomenex, Torrance, CA, USA) was used, with a guard cartridge of the same material. All solvents were filtered through a 0.45 μm filter disk (Millipore Co., Bedford, MA, USA For chestnut shell HPLC mobile phase were: water:formic acid (99.80:0.20, v/v) (A) and methanol (B). Running conditions were: 0 min 95% A and 5% B; 0–45 min 55% B. The system was equilibrated between runs for 10 min using the starting mobile phase composition. The temperature was maintained at 30 °C. Each sample was analyzed three times. The flow-rate was 0.8 mL/min. The injection volume was 20 μL.
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