HPLC波长切换法同时测定藤药中5种成分的含量

目的建立同时测定藤药中5种成分含量的HPLC方法。方法采用Phenomene C18色谱柱(4.6 mm×250 mm,5μm)。流动相采用甲醇(A)-水(B)梯度洗脱:0~15 min(45.0%A),15~27 min(45.0%→56.0%A),27~42 min(56.0%→70.0%A),42~50 min(70.0%→45.0%A)。流速0.8 m L/min;柱温25℃;检测波长:0~15 min(248 nm,α-玉柏碱),15~50 min(254 nm,升麻素苷、升麻素、5-O-甲基维斯阿米醇苷和亥茅酚苷)。结果测得α-玉柏碱、升麻素苷、升麻素、5-O-甲基维斯阿米醇苷和亥茅酚苷的平均加样回收率及对应的RSD分别为98.63%(1.60%)、97.82%(1.45%)、99.18%(1.48%)、97.18%(1.29%)和98.19%(1.80%),且在5.15~103.00、8.13~162.60、4.74~94.80、4.09~81.80、4.35~87.00μg/m L内,线性关系良好(r≥0.999 1)。结论采用HPLC波长切换法能有效分析藤药中的5个成分,方法准确可靠,重复性良好,结果稳定,可以作为藤药产品质量控制的方法。

Simultaneous determination of five constituents in Teng Yao by HPLC wavelength switching method

Objective To develop an HPLC method for simultaneous determination of five components in Teng Yao. Methods The chromatographic analysis was carried out on an Phenomene C18 column ( 4. 6 mm × 250 mm,5 μm). The mobile phase was methanol (A)-water (B) at 0. 8 mL/min in gradient elution mode which was 45. 0% A from 0 to 15 min,45. 0%→56. 0% A from 15 to 27 min,56. 0%→70. 0% A from 27 to 42 min,70% ~45% A from 42 to 50 min. The column temperature was set at 25 ℃,and detection wavelength was set at 248 nm (0~15 min,α-obscurine) and 254 nm (15 ~50 min,prim-o-glucosylcimifugin,cimifugin,5-O-methylvisammioside and sec-o-glucosylhamaudol) . Results Five components were determined by HPLC, including α-obscurine, prim-o-glu-cosylcimifugin,cimifugin,5-O-methylvisammioside and sec-o-glucosylhamaudol. The average recoveries and the corre-sponding RSD were 98. 63% ( 1. 60%) , 97. 82% ( 1. 45%) , 99. 18% ( 1. 48%) , 97. 18% ( 1. 29%) and 98. 19%(1. 80%),respectively. The linearity was good (r≥0. 9991) in the concentration ranges of 5. 15 ~103. 00 μg/mL, 8. 13~162. 60 μg/mL,4. 74 ~94. 80 μg/mL,4. 09 ~81. 80 μg/mL and 4. 35 ~87. 00 μg/mL. Conclusion Five components in Teng Yao are effectively analyzed by HPLC wavelength switching method,and the method is accurate, reproducible with high sensitivity,which can be applied to the quality control of Teng Yao.