ABSTRACT
OBJECTIVE:To establish a method for simultaneous determination of 4 effective components in the couple of Coptidis Rhizoma and Evodiae Fmctus,and to investigate rational proportion of the couple.METHODS:HPLC method was adopted.The determination was performed on Welch XB C18 column with mobile phase consisted of acetonitrile-1.5 mmol/L sodium dodecyl sulfate solution (pH adjusted to 5.0 with phosphoric acid) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 265 nm,and the column temperature was 30 ℃.The sample size was 10 μL.RESULTS:The linear ranges of berberine hydrochloride,martin hydrochloride,evodiamine and rutacarpine were 2.24-44.80 μg/mL(r=0.999 9),1.26-31.50 μg/mL(r=0.999 8),2.70-81.00 μg/mL(r=0.999 8),1.65-49.50 μg mL(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.The recoveries were 98.11％-100.73％ (RSD=1.04％,n=6),96.54％-103.47％(RSD=1.86％,n=6),95.49％-102.36％ (RSD=2.05％,n=6),97.19％-103.24％ (RSD=2.19％,n=6),respectively.When the proportion of Coptidis Rhizoma-Evodiae Fructus(m/m) was 2∶ 1,the comprehensive content of each component was the highest.CONCLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for simultaneous determination of 4 effective components in different compatibility proportions of the couple of Coptidis Rhizoma and Evodiae Fructus.The rational proportion of the couple of Coptidis Rhizoma and Evodiae Fructus is 2 ∶ 1.
ABSTRACT
OBJECTIVE: To determine the concentrations of evodiamine and rutacarpine in subcutaneous tissues of rats after administration of evodiae extract to evaluate the transdermal drug release properties. METHODS: The rats were treated with abdominal hair removal, followed by covering with black polyethylene film transdermal patch of evodiae extract. The concentrations of evodiamine and rutacarpine in the subcutaneous tissues at different time points were measured by microdialysis sampling technique combined with HPLC method. RESULTS: The maximum concentration(ρmax) of evodiamine in the dialysate was(417.24 ± 43.44) μg · mL-1, time to peak concentration (tmax) was (4.210 ± 0.630) h, and the area under the concentration time curve (AUC0→20) was (3 189.31 ± 789.97) μg · h · mL-1; the ρmax of rutacarpine was(287.13 ± 26.78) μg · mL-1, tmax was (3.980 ± 0.580) h, and the AUC0→16 was(l327.97 ± 245.87) μg · h · mL-1. CONCLUSION: In vivo microdialysis sampling technique combined with HPLC method can be used for the evaluation of transdermal drug release characteristics of evodiamine and rutacarpine. The method has the advantages of simple operation with high sensitivity and specificity. Evodiamine and rutacarpine in evodiae extract have the characteristics of slow transdermal penetration speed but high penetration amount.