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目的:建立HPLC法同时测定甜叶菊有效部位中的木犀草素-7-O-β-D-葡萄糖苷、槲皮苷、4,5-咖啡酰奎宁酸和从甜叶菊中分离得到的新化合物槲皮素-3-O-[4-O-反式-咖啡酰基-α-L-鼠李糖-(1→6)-β-D-半乳糖苷]4个酚类成分含量。方法:色谱柱为Hypersil-C18柱(4.6mm×250mm,5μm),流动相为乙腈(A)-0.2%磷酸溶液(B),梯度洗脱[0～30min,A-B(17∶83)→A-B(25∶75);30～40min,A-B(25∶75)→A-B(35∶65)],流速为1.0mL·min-1,检测波长为254nm,柱温:35℃。结果:木犀草素-7-O-β-D-葡萄糖苷、槲皮苷、4,5-双咖啡酰奎宁酸和槲皮素-3-O-[4-O-反式-咖啡酰基-α-L-鼠李糖-(1→6)-β-D-半乳糖苷]的线性范围分别为0.0328～0.656μg(r=0.9996),0.0232～0.464μg(r=0.9996),0.0204～0.408μg(r=0.9997),0.016～0.32μg(r=0.9997)。上述4个化合物平均加样回收率(n=3)均在98.69%～100.9%之间,且RSD均小于3%。结论:本方法可同时测定甜叶菊有效部位中4个酚类成分的含量,结果准确,重复性好。
OBJECTIVE: To establish a HPLC method for simultaneous determination of luteolin-7-O-β-D-glucoside, quercitrin, 4,5-caffeoylquinic acid in the active fractions of Stevia rebaudiana and new The content of quercetin-3-O- [4-O-trans-caffeoyl-α-L-rhamnose- (1 → 6) -β-D-galactoside] METHODS: The chromatographic column was Hypersil-C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of acetonitrile-A 0.2% phosphoric acid solution (B) (25:75); 30 ~ 40min, AB (25:75) → AB (35:65)], the flow rate was 1.0mL · min-1, the detection wavelength was 254nm and the column temperature was 35 ℃. RESULTS: Luteolin-7-O-β-D-glucoside, quercitrin, 4,5-dicaffeoylquinic acid and quercetin-3-O- [4-O-trans-caffeoyl (r = 0.9996), 0.0232 ~ 0.464μg (r = 0.9996), 0.0204 ~ 0.408 μg (r = 0.9997), 0.016-0.32 μg (r = 0.9997). The average recoveries of the four compounds (n = 3) ranged from 98.69% to 100.9% with RSD less than 3%. Conclusion: The method can simultaneously determine the content of four phenolic compounds in the effective parts of Stevia rebaudiana with accurate results and good repeatability.