Chromatographic analysis was conducted following a modified version of the method described previously [24 –26 (link)]. For standardization of RRE, chromatographic analysis was performed using a Hitachi HPLC-DAD system (Hitachi Co., Tokyo, Japan) consisting of a pump (L-2130), autosampler (L-2200), column oven (L-2350), and diode array UV/VIS detector (L-2455). An OptimaPak C18 column (5 μm, 4.6 × 250 mm, RS Tech, Daejeon, Korea) was maintained at 40°C for sample analysis. Output signals from the detector were recorded using EZChrom Elite software from Hitachi. The fingerprinting was conducted with the chemical standards rhapontisterone B, pomolic acid, and echinoynethiophene A (ChemFaces, Wuhan, Hubei, China). The standard solutions were prepared by dissolving each marker component in 100% methanol at 1 mg/mL. RR powder was weighed and dissolved in methanol at 5 mg/mL for analysis. The standard solutions and sample were injected at a volume of 10 μL and were detected at a UV wavelength of 254 nm. The mobile phase was water with 0.1% acetic acid (A) and acetonitrile with 0.1% acetic acid (B) at a flow rate of 1.0 mL/min with the following gradient flow: 10% B at 0–10 min, 10–50% B at 10–50 min, 50% B at 50–55 min, and 50–100% B at 55–60 min. The identification of the peaks was based on the UV spectrum and retention time of each marker component from the RRE.
Ezchrom elite software
Manufactured by Hitachi
Sourced in Japan
EZChrom Elite software is a data acquisition and analysis tool designed for chromatographic applications. It provides essential functionality for managing and processing chromatographic data from various analytical instruments.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using ezchrom elite software
1
Fingerprinting of Rhaponticum Carthamoides Extract
2
Amino Acid Analysis for Protein Quantitation
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Amino acid analysis was used to determine accurate total protein concentrations of each of the chloroform/methanol-extracted samples so that equal amounts of protein could be digested and subjected to DDA, DIA, and PRM analysis. Briefly, an aliquot of each serum sample was dried in a Speed-Vac and then hydrolyzed for 16 hours at 115°C in 100 μL of 6 N HCl, 0.2% phenol with 2 nmol norvaline as an internal standard that was used to correct for losses that may occur during sample transfers, drying, and so on. After hydrolysis, the acid was removed by drying in a Speed-Vac, and the resulting amino acids were dissolved in 100 μL loading buffer (0.02 N HCl) with 2 nmol taurine as a second internal standard that was used to independently monitor transfer of the sample onto the analyzer. The analyses were carried out on a Hitachi L-8900A Amino Acid Analyzer with ion-exchange separation of the amino acids and post-column derivatization with ninhydrin for detection. Data were collected and analyzed with the Hitachi EZChrom Elite software.
3
Analytical HPLC of Organic Compounds
4
Soybean Nutrient Composition Analysis
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Fresh leaves were randomly collected on the 35th day after the sowing of soybean seeds (G3, R1, SC) for the measurement of nutrient content. Half a gram of fresh leaflets and phosphate-buffered saline (PBS, pH 8.0) was placed in a mortar, ground with a pestle in an ice-water bath, and centrifuged at 2500× g for 10 min. The supernatant was used as the test solution. The composition and content of amino acids in soybeans were determined by a Hitachi amino acid analyzer using EZChrom Elite software (L-8900, Hitachi Limited, Japan). The content of soluble protein and soluble sugar in soybeans was determined using a total protein quantification kit and plant soluble sugar content test kit) from Qingdao Sci-Tech Innovation Quality Testing Co. LTD (Qingdao, China). The optical density (OD) of the reactants was measured using an ultraviolet spectrophotometer (UV-1800PC, Mapada; Shanghai, China). Three replicates per soybean variety were set up.
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