磷脂水溶液制备方法的研究

Preparation Method of Phospholipid-Aqueous Solution

大豆磷脂具有多种生物活性,但其不溶于水的特性限制了细胞生物学功效的研究。以丙三醇、二甲基亚砜(DMSO)和无水乙醇作为磷脂的溶解剂,探究大豆磷脂细胞培养液的制备方法。首先,采用酶标仪全波长扫描并缩小波长范围确定大豆磷脂最佳线性吸收波长;其次,以正己烷溶解大豆磷脂并制备标准曲线,研究其在3种溶剂中的溶解度,并检测大豆磷脂-甘油-水溶液中溶质颗粒粒径;最后,采用MTT方法检测磷脂细胞培养液对Caco-2细胞的影响。结果表明:当波长为340nm时,大豆磷脂的吸光度与质量浓度之间呈最佳线性关系;大豆磷脂在无水乙醇、DMSO、丙三醇中的溶解量分别为28.8、31.6、180mg/mL;含大豆磷脂的溶液对Caco-2细胞的增殖有明显促进作用,随质量浓度的升高,促进作用增强。因此,丙三醇可作为大豆磷脂的溶解剂,配成水溶液用于大豆磷脂细胞功效的检测。

Soybean phospholipids have many bioactivities.However,the poor solubility of soybean phospholipids in water limits its cellular biological efficacy.In this work,glycerol,dimethylsulphoxide(DMSO)and ethanol were used as the solvents to dissolve phospholipids and to explore the preparation method of soybean phospholipids in cell culture solution.Firstly,the optimal linear absorption wavelength was obtained by the whole wavelength scan and gradually reducing the wavelength range with microplate reader.Secondly,the solubility of soybean phospholipids in three solvents were investigated according to the standard curve of soybean phospholipids in hexane solution,and the particle size of soybean phospholipid-glycerol-aqueous solution was measured by Malvern Particle.Finally,the effect of soybean phospholipid-glycerol solution on the Caco-2 cells was detected by MTT assay.The results showed that at 340 nm,the absorbance of soybean phospholipid had the best linear relationship with its concentration.The solubilities of soybean phospholipid in ethanol,DMSO and glycerol were 28.8 mg/mL,31.6 mg/mL and 180 mg/mL,respectively.The effect of soybean phospholipid-propanol solution on the Caco-2 cells was enhanced with the increase of its concentration.Therefore,glycerol was used as the soluble agent for soybean phospholipids in aqueous solution,which wald be used for the determination of soybean phospholipids in cell culture.