目的: 测定银屑病外用方中7个主要活性成分(新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷、柠檬苦素)含量。方法:采用Waters Xbrige BEH C18色谱柱(150 mm×4.6 mm, 3.5 μm),以水(A)-乙腈(B)为流动相,梯度洗脱(0~5 min,17%B;5~10 min,17%B→20%B;10~15 min,20%B;15~25 min,20%B→100%B),流速1.0 mL·min-1,柱温30 ℃,进样量5 μL,检测波长为292 nm(0~19 min,新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷)、210 nm(19~25 min,柠檬苦素),使用MATLAB软件编程,对CSV格式数据进行双波长融合。结果:新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷和柠檬苦素线性范围分别为39.06～1 250 μg·mL-1(r=0.999 9)、26.25~840 μg·mL-1(r=0.999 7)、18.64～596.4 μg·mL-1(r=0.999 8)、6.31～202 μg·mL-1(r=0.999 6)、2.53～80.8 μg·mL-1(r=0.999 8)、1.33～42.6 μg·mL-1(r=0.999 7)和5.08～162.4 μg·mL-1(r=0.999 6),平均加样回收率(n=6)为96.1%~102.2%,RSD为0.48%~1.7%。银屑病外用方自制及医院提供的样品中新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷和柠檬苦素的含量分别为1.06、1.21、0.87、0.30、0.09、0.07、0.22 mg·mL-1和0.35、0.32、0.17、0.08、0.04、0.03、0.21 mg·mL-1。结论:该方法能够快速、准确地测定银屑病外用方中7个活性成分含量,可用于银屑病外用方的质量控制。
Objective: To determine the contents of 7 active components(neoastilbin, astilbin, neoisoastilbin, isoastilbin, engeletin, isoengeletin, limonin) in the psoriasis external prescription. Methods: The column was Waters Xbrige BEH C18(150 mm×4.6 mm, 3.5 μm). The mobile phase consisted of solvent A(water) and solvent B(acetonitrile) with gradient elution (0-5 min, 17%B; 5-10 min, 17%B→20%B; 10-15 min, 20%B; 15-25 min, 20%B→100%B). The flow rate was 1.0 mL·min-1. The column temperature was 30 ℃. Injection volume was 5 μL. The detection wavelength for neoastilbin, astilbin, neoisoastilbin, isoastilbin, engeletin and isoengeletin were set at 292 nm during 0~19 min, and 210 nm for limonin during 19~25 min. MATLAB was used for two-wavelength fusion of CSV data. Results: The calibration curves of neoastilbin, astilbin, neoisoastilbin, isoastilbin, engeletin, isoengeletin and limonin were all in good linearity over the ranges of 39.06-1 250 μg·mL-1(r=0.999 9), 26.25-840 μg·mL-1(r=0.999 7), 18.64-596.4 μg·mL-1(r=0.999 8), 6.31-202 μg·mL-1(r=0.999 6), 2.53-80.8 μg·mL-1(r=0.999 8), 1.33-42.6 μg·mL-1(r=0.999 7) and 5.08-162.4 μg·mL-1(r=0.999 6), respectively, and the average recoveries(n=6) were 96.1%~102.2% with RSD of 0.48%-1.7%. The contents of neoastilbin, astilbin, neoisoastilbin, isoastilbin, engeletin, isoengeletin and limonin in samples self-made and provided by a hospital were 1.06, 1.21, 0.87, 0.30, 0.09, 0.07, 0.22 mg·mL-1 and 0.35, 0.32, 0.17, 0.08, 0.04, 0.03, 0.21 mg·mL-1, respectively. Conclusion: This established method can determine the contents of 7 active ingredients in psoriasis external prescription quickly and accurately, and is also helpful for the quality control of psoriasis external prescription.
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