HPLC analysis was carried out according to a previously described method [14 (link)] with few modifications. In brief, aqueous extract was fractionated with ethyl acetate and was dissolved in methanol (1 mg/mL) before subjecting to analysis. The HPLC system (Shimadzu, Japan) consisted of dual pump LC-20AD binary system, PDA detector SPD-M20A; Merck C18 reversed-phase column (I.D. 4.6 mm × 250 mm, 5 μm). Separation was carried out using a two-pump linear gradient program for pump A (40% acetonitrile) and pump B (5% acetonitrile). Elution was initiated with a gradient of 12% A changing to 40% in 20 min, 100% in 45 min and finally to 12% in 55 min followed by washing for 35 min. Injection volume was 20 μL and flow rate was maintained at 1 mL/min throughout the process.
Lc 20ad binary system
Manufactured by Merck Group
Sourced in Japan
The LC-20AD binary system is a laboratory equipment product manufactured by Merck Group. It is a high-performance liquid chromatography (HPLC) system that utilizes two independent solvent delivery pumps to create a binary gradient for sample separation and analysis.
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2 protocols using lc 20ad binary system
1
HPLC Analysis of Plant Extracts
2
HPLC Analysis of Phytoconstituents
Check if the same lab product or an alternative is used in the 5 most similar protocols
The presence of phytoconstituents, β-sitosterol, lupeol and quercetin were investigated by HPLC using Waters HPLC instrument (Kyoto, Japan) fitted with a dual pump LC-20AD binary system with photodiode array (PDA) detector SPD-M20A, Merck RPC18 column (I.D. 4.6 x 250 mm, 5 mm). MCU was dissolved in methanol and injected. Gradient elution was carried out with methanol: phosphate buffer (50 M) at pH 3, (70:30) and the flow rate was adjusted to 1.0 mL/min with 20 µL injection volume, detection by UV at 250 nm.
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The presence of phytoconstituents, β-sitosterol, lupeol and quercetin were investigated by HPLC using Waters HPLC instrument (Kyoto, Japan) fitted with a dual pump LC-20AD binary system with photodiode array (PDA) detector SPD-M20A, Merck RPC18 column (I.D. 4.6 x 250 mm, 5 mm). MCU was dissolved in methanol and injected. Gradient elution was carried out with methanol: phosphate buffer (50 M) at pH 3, (70:30) and the flow rate was adjusted to 1.0 mL/min with 20 µL injection volume, detection by UV at 250 nm.
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