柱前衍生化高效液相色谱法测定荞麦中D-手性肌醇含量的方法

Method for the Content Determination of D-Chiro-Inositol from Buckwheat by Pre-column Derivatization HPLC

建立了一种柱前衍生化高效液相色谱法(HPLC)测定荞麦中D-手性肌醇(D-CI)含量的方法,对衍生化试剂及方法的重复性、稳定性、精密性及加标回收率进行了考察。得到衍生化条件为:苯甲酰氯0.2 m L,吡啶0.6 m L,反应温度70℃,反应时间60 min;色谱条件为:C18柱(4.6 mm×250 mm,5μm),以乙腈和超纯水为流动相进行梯度洗脱,流速为1.0 m L/min,柱温30℃,UV检测波长230 nm。结果表明,荞麦中的D-CI获得了很好的分离,D-CI在0.004~0.04 mg/m L内线性相关性好,R^2为0.999,加标回收率为98.40%~100.67%,所建立的方法稳定、灵敏、重现性好,可用于测定荞麦中D-CI含量定量分析。

A method was established for the determination of D- chiro- inositol （ D- CI ） by pre- column derivatization HPLC. Repeatability, stability, precision and added standard recovery of the method and the derivatization conditions were investigated.Derivatization conditions were as follows：0.2 mL of benzoyl chloride,0.6 mL of pyridine, reaction temperature 70 ℃ , reaction time of 60 min.Chromatographic conditions were as follows：using C18 column （4.6mmx250mm,5μm） , with acetonitrile and ultrapure water as mobile phrase in gradient elution, flow rate of 1.0 mL/min, column temperature of 30 ℃, and UV detection wavelength of 230 nm.The results showed that D-CI of buckwheat won good separation by the method, and the method got a good linear relationship of D-CI concentration and peak area at a range of 0.004-0.04 mg/mL with 0.999 of R2 and the recovery was 98.40%- 100.67% .So the method was stable, sebsitive, good reproducibility, and could be used to determine the content of D-CI from buckwheat.