School of Light Industry and Chemical Engineering, Dalian Polytechnic University, Dalian 116034, China.
College of Marine Science and Environment, Dalian Ocean University, Dalian 116023, China.
J Chromatogr Sci. 2023 Oct 3;61(8):737-742. doi: 10.1093/chromsci/bmac051.
A high-speed counter-current chromatographic (HSCCC) method using ethyl acetate-water as solvent system was established to separate isoquercetin from Toona sinensis. In an HSCCC single separation, the sample sizes of ethanol extract were optimized from 203 to 1200 mg. The results showed that the yield of the target compound increased from 4 to 26 mg, and the corresponding purity decreased from 93.30 to 81.82%. To further improve the yield and purity, liquid-liquid-refining extraction was introduced to pretreat the ethanol extract and enrich the target compound. The ethanol extract was extracted with n-hexane-ethyl acetate-method-water (1:5:1:5, v/v) and ethyl acetate-water in turn to remove the low-polarity and high-polarity impurities and obtain the crude sample. Under the similar conditions, 85.25 mg of the target compound with the purity of 95.12% was separated from 240 mg of the crude sample. Subsequently, a consecutive HSCCC was developed to obtain 257 mg of the target compound from 720 mg of crude sample, which was equivalent to 14.4 g of ethanol extract. This method improved the purity of the target compound, but more importantly, the sample size can reach 12 times of the maximum sample size of the ethanol extract in a single run.
建立了一种采用乙酸乙酯-水为溶剂体系的高速逆流色谱（HSCCC）方法，从香椿中分离异槲皮苷。在 HSCCC 单次分离中，优化了乙醇提取物的样品量从 203 至 1200 毫克。结果表明，目标化合物的收率从 4 增加到 26 毫克，相应的纯度从 93.30 降低到 81.82%。为了进一步提高产率和纯度，采用液-液精制萃取法预处理乙醇提取物并富集目标化合物。将乙醇提取物依次用正己烷-乙酸乙酯-水（1:5:1:5，v/v）和乙酸乙酯-水萃取，以去除低极性和高极性杂质，并获得粗样品。在相似的条件下，从 240 毫克粗样品中分离出 85.25 毫克纯度为 95.12%的目标化合物。随后，开发了连续 HSCCC，从 720 毫克粗样品中分离出 257 毫克目标化合物，相当于 14.4 克乙醇提取物。该方法提高了目标化合物的纯度，但更重要的是，单次运行的样品量可达到乙醇提取物最大样品量的 12 倍。
