The leaves of A. fruticosa L. used in this study were collected from Jiaxian, Shaanxi Province, China. Samples for isolation were harvested in June 2013. Leaves for quantitative analysis were collected from the same plants on May 20, June 30, and August 10, 2014, and samples at each sampling time were collected from three plants in a wild field. The sample collected on May 20 was named AL0520, and the remaining samples were also named the same way. Their botanical origins were identified by the corresponding author (Naisheng Bai), and a voucher specimen (AF-2013-01) has been deposited in Room 612, Department of Pharmaceutical Engineering, College of Chemical Engineering, Northwest University, Xi’an, China. Column chromatography was performed over silica gel (200–300 mesh; Qingdao Marine Chemical Factory, Qingdao, China), polyamide, MCI GEL CHP-20P, and Sephadex LH-20 (Sigma-Aldrich, St. Louis, MO, USA). Precoated silica gel 60 GF254 plates were supplied by Merck (Darmstadt, Germany).
Mci gel chp 20p
Manufactured by Merck Group
Sourced in China, Sweden, United States
The MCI GEL CHP-20P is a lab equipment product. It is a type of chromatography column used for the purification and separation of chemical compounds. The device operates using principles of high-performance column chromatography.
Automatically generated - may contain errors
Lab products found in correlation
Spectramax m3 multi mode microplate reader, by Molecular Devices (2 mentions)
Sephadex lh 20, by Merck Group (1 mentions)
Precoated silica gel 60g f254 plates, by Merck Group (1 mentions)
C18 column, by Dikma Technologies (1 mentions)
Agilent 1200 system, by Agilent Technologies (1 mentions)
Lc forte r 100, by YMC (1 mentions)
Jms 700, by JEOL (1 mentions)
Sephadex lh 20, by Cytiva (1 mentions)
Precoated rp 18 f254s plates, by Merck Group (1 mentions)
Lichroprep rp 18, by Merck Group (1 mentions)
P 2000 digital polarimeter, by Jasco (1 mentions)
Aq c18, by YMC (1 mentions)
Acclaim polar advantage 2 c 18, by Thermo Fisher Scientific (1 mentions)
Du650 spectrophotometer, by Beckman Coulter (1 mentions)
5 protocols using mci gel chp 20p
1
Isolation and Analysis of Fruticosa Leaf Compounds
2
Metabolite Extraction and HPLC Analysis of Physcomitrella patens
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The gametophores of P. patens were fully ground in liquid nitrogen. The sample (0.3 g) was dissolved with 5 mL methanol. After centrifugation (4,000 rpm, 10 min) of the extract, the supernatant was concentrated and rinsed with 4 mL 0.2% acetic acid containing cyclohexane (1:1, v/v). The mixture was subjected to MCI GEL CHP 20P (Sigma-Aldrich) resin solid-phase extraction column, and ethanol was used as eluent. The extraction was filtered with a 0.22 μm ultrafiltration membrane (Pall Corp., Port Washington). Then the analysis was carried out by high-performance liquid chromatography (HPLC) equipped with a photodiode array detector (Agilent 1,200 system). The experiment was under the following conditions: C18 column (4.6 mm×250 mm, 5 μm, Dikma Technologies, Beijing); injection volume, 20 µL; column temperature, 35 °C; flow rate, 1.0 mL/min; solvent system, acetonitrile (A): 0.2% acetic acid (B) with gradient elution. The gradient conditions were 3% A for 6 min, then a linear gradient to 90% over 65 min, and the detection wavelength was 280 nm.
3
Isolation and Purification of Compounds
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Used chemicals
were of analytical grade and were bought from Thermo Fisher Scientific
(Waltham, Massachusetts, USA). Octadecylsilanized (ODS) silica gel
(YMC Ltd., Japan), Sephadex LH-20 (Amersham Pharmacia Biotech, Sweden),
and MCI GEL CHP20P (Sigma-Aldrich Korea, Seoul, South Korea) were
used as packing materials for column chromatography. Recycling HPLC
(LC-Forte/R100, YMC Co., LTD, Kyoto, Japan) system arranged with a
UV detector was used for the isolation and purification of compounds.
NMR spectra of the purified compounds were conducted on a Bruker (AM
300, 500 MHz) and JEOL JMS-700 (JEOL Ltd., Akishima, Japan) was used
for the MS. The CD spectra were recorded on a Jasco J-815 (Japan)
CD spectrophotometer. The enzyme inhibition assays were evaluated
on a SpectraMaxM3Multi-Mode Microplate Reader (Molecular device, USA).
were of analytical grade and were bought from Thermo Fisher Scientific
(Waltham, Massachusetts, USA). Octadecylsilanized (ODS) silica gel
(YMC Ltd., Japan), Sephadex LH-20 (Amersham Pharmacia Biotech, Sweden),
and MCI GEL CHP20P (Sigma-Aldrich Korea, Seoul, South Korea) were
used as packing materials for column chromatography. Recycling HPLC
(LC-Forte/R100, YMC Co., LTD, Kyoto, Japan) system arranged with a
UV detector was used for the isolation and purification of compounds.
NMR spectra of the purified compounds were conducted on a Bruker (AM
300, 500 MHz) and JEOL JMS-700 (JEOL Ltd., Akishima, Japan) was used
for the MS. The CD spectra were recorded on a Jasco J-815 (Japan)
CD spectrophotometer. The enzyme inhibition assays were evaluated
on a SpectraMaxM3Multi-Mode Microplate Reader (Molecular device, USA).
4
NMR Characterization of Organic Compounds
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The NMR spectra were obtained at 400 and 100 MHz for 1H and 13C, respectively, on a Varian MR 400 NMR spectrometer (Agilent Technologies, Santa Clara, CA, USA) in CDCl3 or pyridine-d5, with solvent peaks as references. Silica gel 60 (Merck Millipore, 230–400 mesh), MCI gel CHP20P (Merck Millipore, 75–150 μm), and LiChroprep RP-18 (Merck Millipore, 40–63 μm) were used for column chromatography. Precoated silica gel plates (Merck Millipore, Kieselgel 60 F254, 0.25 mm) and precoated RP-18 F254s plates (Merck Millipore, 1.05560) were employed in TLC analyses. Spots were visualized under ultraviolet (UV) light or heating silica gel plates sprayed with 10% H2SO4 in CH3OH. High-performance liquid chromatography (HPLC) was performed on a Primaide 1110 pump equipped with a Primaide 1410 UV detector at 220 nm and a semipreparative reversed-phase column (Merck Millipore, Hibar Purospher RP-18e, 5 μm, 250 mm × 10 mm).
5
Phytochemical Analysis of A. fruticosa
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Open column chromatography was carried out using MCI GEL CHP20P (75–150 μm, Merck, Kenilworth, NJ, USA). Preparative recycling medium pressure liquid chromatography (MPLC) and HPLC were conducted on an LC-9130G NEXT (Jai Co., Ltd., Tokyo, Japan) using AQ C18 (S-10 μm, 12 nm, YMC, Kyoto, Japan) and Acclaim Polar Advantage II C-18 (S-5 μm, 12nm, Thermo Fisher Scientific, Waltham, MA, USA). One-dimensional, as well as 2D NMR spectra, were recorded by a Bruker AM500 spectrometer (Billerica, MA, USA), using either methanol-d4 or acetone-d6 as solvent and tetramethylsilane (TMS) as an internal standard. UV spectra were obtained using a DU650 spectrophotometer (Beckman Coulter, Brea, CA, USA). HRESIMS were conducted using Vion (Waters, Milford, MA, USA). Enzymatic assays were implemented using a SpectraMax M3 Multi−Mode microplate reader (Molecular Devices, San Jose, CA, USA). The specific rotation was estimated using a P−2000 Digital Polarimeter (JASCO, Tokyo, Japan). All chemicals for analysis were of first grade. The roots of A. fruticosa were harvested at Naedong in Jinju, South Korea, in June 2017.
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