石榴果实没食子酸及其合成相关物质HPLC 测定体系优化

摘要

建立了‘泰山三白甜’石榴果实不同部位没食子酸及其合成相关物质的高效液相色谱（HPLC）定性和定量检测方法。色谱条件：色谱柱为 Agilent Zorbax XDB －C18（4．6 mm ×150 mm，5μm），没食子酸以乙腈－2％冰乙酸水溶液为流动相（20∶80），检测波长270 nm；莽草酸和3－脱氢莽草酸以甲醇－1％磷酸水溶液（5∶95）为流动相，检测波长214 nm；五没食子酰葡萄糖以乙腈－0．5％磷酸水溶液（20∶80）为流动相，检测波长275 nm，4种物质均等度洗脱，流速均为0．8 mL／min，柱温均为30℃。结果表明，4种物质在一定线性范围内的峰面积与质量浓度呈良好的线性关系，相关系数均大于0．99，检测限为0．12～0．24 mg／L；加标回收率为98．9％～102．5％，相对标准偏差（RSD）为1．2％～2．2％。‘泰山三白甜’石榴果实不同部位没食子酸、莽草酸、3－脱氢莽草酸和五没食子酰葡萄糖含量存在显著差异，果皮中含量最高，果汁次之，种子中最低。该方法操作简单，灵敏度高，重复性好，可用于没食子酸及其合成相关物质的快速鉴定及含量测定。%The qualitative and quantitative determination methods of gallic acid and its synthetic related substances in different parts of Taishansanbaitian pomegranate fruit were developed by high performance liquid chromatography (HPLC).The chromatographic column was Agilent Zorbax XDB -C1 8 (4.6 mm ×1 50 mm, 5 μm).The mobile phase of gallic acid was acetonitrile and 2% glacial acetic acid solution with the ratio of 20∶80.Its determine wavelength was 270 nm.The mobile phase of shikimic acid and 3 -dehydroshikimic acid was methanol and 1 % phosphoric acid with the ratio of 5∶95.Their determine wavelengths both were 21 4 nm.The mobile phase of pentagalloylglucose was acetonitrile and 0.5% phosphoric acid with the ratio of 20∶80.Its determine wavelength was 275 nm.The 4 substances were all isocratic elution with flow rate of 0.8 mL/min and column temperature of 30℃.The results indicated that the peak area and mass concentration of 4 substances presented good linearities in certain ranges.The correlation coefficients of 4 substances were all o-ver 0.99.The detection limits were 0.1 2 ~0.24 mg/L.The standard recovery rates were 98.9% ~1 02.5%. The relative standard deviationas (RSDs)were 1 .2% ~2.2%.The contents of gallic acid,shikimic acid, 3 -dehydroshikimic acid and pentagalloylglucose were significantly different in different parts of Taishansan-baitian pomegranate fruit.The contents were the highest in fruit peel,followed by juice and seed.The determi-nation method was simple,and had high sensitivity and good reproducibility,which could be used for rapid i-dentification and determination of gallic acid and its synthetic related substances in pomegranate fruit.