SPE-HPLC-ELSD法测定复方丹参浸膏中五种低聚糖成分的方法研究

Determination of Five Oligosaccharide in Compound Danshen Extract by SPE-HPLCELSD

目的建立一种以固相萃取-高效液相色谱-蒸发光散射法（SPE-HPLC-ELSD）对复方丹参浸膏中的五种低聚糖成分进行测定的方法。方法使用Prevail TM Carbohydrate ES色谱柱,以乙腈和水为流动相,梯度洗脱（0-23 min,乙腈75%→60%）,柱温30℃,流速为0.8 ml·min^-1;ELSD漂移管温度60℃,载气为空气,气压25 psi。结果复方丹参浸膏中的五种低聚糖成分能够得到较好的分离,葡萄糖、果糖、蔗糖、Glu-Fru-Gal三糖和Gal-Gal-Glu-Fru四糖的线性范围分别为0.246-1.230,0.236-1.182,0.246-1.230,0.068 2-1.364和0.580 5-5.805 0 mg;平均加样回收率（n=6）分别为101.7%（RSD=1.6%）、102.0%（RSD=1.0%）、100.8%（RSD=0.9%）、102.8%（以棉子糖计,RSD=1.2%）和98.5%（RSD=1.8%）。结论该方法快速简捷、结果准确稳定,适合于复方丹参浸膏中的五种低聚糖成分的含量测定。

Objective An SPE-HPLC-ELSD method was established for the quantitative analysis of the oligosaccharide in compound Danshen extract. Methods The chromatographic separation was achieved on Prevail TM Carbohydrate ES column using a mobile phase composed of a mixture of acetonitrile and water by gradient elution（0 - 15 min,acetonitrile 75%→60%） at a flow rate of 0.8 ml · min^-1. The ELSD drift tube temperature should be set at 60℃ and the carrying gas was air. Results Under the conditions above, five oligosaccharide could be well separated, and the linear ranges of fructose,glucose, sucrose, Glu–Fru-Gal and Gal-Gal-Glu-Fru were 0.246 - 1.230 、0.236 - 1.182、0.246 - 1.230 、0.068 2 - 1.364 and 0.580 5 - 5.805 0 mg; The average spiked recoveries（n = 6） were 103.0%（RSD = 2.4%）、103.3%（RSD = 2.3%）、102.4%（RSD= 2.3%）、102.8%（ RSD = 1.2%） and 98.5%（RSD = 1.8%）. Conclusion This method is accurate, rapid, simple and reproducible, and thus it is suitable for the determination of oligosaccharide in compound Danshen extract.