HPLC和UPLC法测定复方黄连素片中吴茱萸碱、吴茱萸次碱含量的方法比较

Comparison of HPLC and UPLC methods for determination of evodiamin and rutacarpine in compound Huangliansu tablets

目的:比较测定复方黄连素片中吴茱萸碱和吴茱萸次碱含量的高效液相色谱法(HPLC)与超高效液相色谱法(UPLC)。方法:HPLC色谱条件:采用Atlantis C18(4.6 mm×250 mm,5μm)色谱柱,以10 mmol.L-1醋酸铵(pH 3.5)为流动相A,乙腈为流动相B,梯度洗脱,流速1 mL.min-1,检测波长240 nm,柱温35℃;UPLC色谱条件:Waters Acquity HSS T3(2.1 mm×100 mm,1.7μm)色谱柱,以10 mmol.L-1醋酸铵(pH 3.5)为流动相A,以乙腈为流动相B,梯度洗脱,流速0.5 mL.min-1,检测波长240 nm,柱温35℃。结果:HPLC测定,吴茱萸碱和吴茱萸次碱线性范围分别为0.425～5.30μg和0.409～5.110μg(r>0.999),平均回收率(n=9)分别为98.6%和99.0%;UPLC测定,吴茱萸碱和吴茱萸次碱线性范围分别为0.106～0.743μg和0.102～0.715μg(r>0.999),平均回收率(n=9)分别为99.1%和99.2%。结论:UPLC较HPLC更加简便迅速,既达到了分析的高通量,得到更高的分析灵敏度,又大大提高了分析速度,减少了有机溶剂的消耗。UPLC能够成功替代HPLC,为复方黄连素片中吴茱萸药材提供新的质量控制方法。

Objective： To compare HPLC and UPLC methods in the determination of evodiamin and rutacarpine in compound Huangliansu tablets. Methods： HPLC condition： Atlantis C18 column （4. 6 mm × 250 mm, 5 μm） , 10 mmol L-1 ammonium acetate （pH 3.5 ） as mobile phase A, acetonitrile as mobile phase B, gradient elution, ve- locity at 1 mL min- 1, detected wavelength of 240 nm, column temperature at 35 ℃ ;UPLC condition：Waters Ac- quity HSS T3 column（2. 1 mm × 100 mm,1.7 μm） ,10 mmol L-1 ammonium acetate（pH 3.5） as mobile phase A, acetonitrile as mobile phase B, gradient elution, velocity at 0.5 mL min - l, detected wavelength of 240 nm, col- umn temperature at 35 ℃. Results： HPLC determination： the linear ranges of evodiamin and rutacarpine were 0. 425 -5.30 μg and 0. 409 -5. 110 μg（r 〉0. 999） ,respectively;the average recovery（n =9） were 98.6% and 99.0% ,respectively. UPLC determination：the linear ranges of evodiamin and rutacarpine were 0. 106 -0. 743 μg and 0. 102 - 0.715 μg （ r 〉 0. 999 ）, respectively ; the average recovery （ n = 9 ） were 99. 1% and 99.2 %, respective- ly. Conclusion： Comparing with HPLC method, UPLC method is relatively quicker, with higher throughput, higher sensitivity and less organic solvent. UPLC method can succeed alternative HPLC for Fructus Evodiae quality control in compound Huangliansu tablets.