A. distichum leaves were obtained from Woorinamoo Agricultural Union Corporation (Goesan, Korea). The collected leaves were then processed and extracted at Korea Prime Pharmacy Co., Ltd. (Gwangju, Korea). Two kilograms of the leaves were dried, crushed and extracted with 40 L of 70% ethanol, for 4 h at 75°C. After filtration using Whatman filter paper (No. 4, Whatman International Ltd., Kent, United Kingdom), the solvent was evaporated and lyophilized to obtain 1.02 kg of ALE extract which was stored at −20°C until use. The samples were analyzed by HPLC on a Shimadzu liquid chromatography system LC-20AD using a diode array detector (SPD-M20A) and YMC-Pack ODS-AM column (4.6 × 250 mm, 2.6 μm). Solvents A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile) were used as mobile phase. The run time was 30 min with a flow rate of 1 mL/min, detected at 330 nm and the concentration was calculated comparing the sample peak area to that of standards. The results obtained is given in Supplementary Fig. 1 .
Lc 20ad
Manufactured by YMC
Sourced in Japan
The LC-20AD is a liquid chromatography system designed for analytical-scale separations. It features a dual-plunger parallel-flow pump capable of generating high pressures, enabling the analysis of a wide range of sample types. The system is equipped with advanced control and data management software for reliable and precise operation.
Automatically generated - may contain errors
Lab products found in correlation
Ods am column, by YMC (1 mentions)
Whatman filter paper, by Cytiva (1 mentions)
Atractylenolide 3, by Merck Group (1 mentions)
Atractylenolide 1, by Merck Group (1 mentions)
Spd 20a, by YMC (1 mentions)
Triart c18 column, by YMC (1 mentions)
Lc dad system, by Shimadzu (1 mentions)
Pro c18 column, by YMC (1 mentions)
3 protocols using lc 20ad
1
Extraction and Analysis of Adhatoda Leaves
2
Atractylenolide Analysis by HPLC
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Samples were analyzed by a reverse-phase HPLC system (Shimadzu 20A, Kyoto, Japan) that consisted of an autosampler (SIL-20A), a binary pump (LC-20AD), and a photodiode array detector (SPD-20A) and was equipped with a YMC-Triart C18 column (5 μm × 4.6 mm × 250 mm) (YMC, Kyoto, Japan). Gradient flows for the two-solvent system (solvent A, 0.05% phosphoric acid in water; solvent B, acetonitrile) were as follows: 85% A/15% B at 0 min, 85% A/15% B at 5 min, 50% A/50% B at 15 min, 50% A/50% B at 20 min, 40% A/60% B at 25 min, 40% A/60% B at 30 min, 15% A/85% at 35 min, 15% A/85% at 40 min, 85% A/15% at 42 min, and 85% A/15% at 45 min. The flow rate of the mobile phase was 1.0 ml/min with an injection volume of 10 μl. Detection was performed at 220 nm for atractylenolide III (Sigma, St. Louis, MO, USA) or at 280 nm for atractylenolide I (Sigma).
3
Analytical and Preparative HPLC Purification
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Analytical HPLC was carried out using a Shimadzu LC-DAD system (Shimadzu, Kyoto, Japan; Degaser: DGU-20A3R prominence degassing Unit, Liquid Chromatograph: LC-20 AD prominence liquid chromatograph, SIL-20A HAT prominence autosampler, and SPA-M20A prominence diode-array detector) with a YMC-Pack Pro C18 column (S-5 µm, 12 nm, 150 × 10.0 mm). The preparative HPLC was performed according to Buckett et al. (2020) [43 (link)]. Regarding the biosynthesised compounds, each peak was collected using the analytical method and, after approximately 20 runs, this resulted in enough material for NMR.
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