多波长HPLC梯度洗脱法同时测定眠安宁合剂中7个有效成分含量

Simultaneous Determination of Seven Active Components in Mian’anning Mixture by HPLC Under Multiple UV Wavelengths

关键词:  眠安宁合剂  5-羟甲基糠醛  远志(口山)酮Ⅲ  白术内酯Ⅰ  白术内酯Ⅲ  丹酚酸B  丹参酮ⅡA  梓醇

Key words:  Mian’anning mixture  5-hydroxymethyl-2-furaldehyde  polygalaxanthone Ⅲ  atractylenolide I  atractylenolide Ⅲ  salvianolic acid B  tanshinone ⅡA  catalpol

摘要:

目的 建立多波长高效液相梯度洗脱法同时测定眠安宁合剂中5-羟甲基糠醛、远志(口山)酮Ⅲ、白术内酯Ⅰ、白术内酯Ⅲ、丹酚酸B、丹参酮ⅡA、梓醇7个成分的含量。方法 采用依利特Hypersil C₁₈色谱柱(4.6 mm×250 mm，5 μm)；流速：0.7 mL·min^-1；流动相A为甲醇-乙腈(1∶3)，流动相B为0.1%磷酸水溶液，梯度洗脱；检测波长：5-羟甲基糠醛为284 nm，远志(口山)酮Ⅲ为320 nm，白术内酯Ⅰ和白术内酯Ⅲ为220 nm，丹酚酸B和丹参酮ⅡA为270 nm，梓醇为210 nm，柱温为25 ℃。结果 5-羟甲基糠醛、远志(口山)酮Ⅲ、白术内酯Ⅰ、白术内酯Ⅲ、丹酚酸B、丹参酮ⅡA和梓醇质量浓度分别在10.08~100.80 μg·mL^-1(r=0.999 8)、3.55~35.50 μg·mL^-1(r=0.999 3)、5.80~58.00 μg·mL^-1(r=0.999 1)、4.20~42.00 μg·mL^-1(r=0.999 9)、40.50~405.00 μg·mL^-1(r=1.000)、12.28~122.80 μg·mL^-1(r=0.999 5)、7.90~79.00 μg·mL^-1(r=0.999 9)内与其峰面积呈良好的线性关系；5-羟甲基糠醛、远志(口山)酮Ⅲ、白术内酯Ⅰ、白术内酯Ⅲ、丹酚酸B、丹参酮ⅡA和梓醇的平均加样回收率分别为98.56%，96.94%，99.61%，97.54%，100.04%，98.59%，99.81%，RSD分别为0.97%，1.73%，1.03%，1.35%，0.86%，1.70%，1.43%(n=6)。结论 建立的方法准确可靠，可用于眠安宁合剂的含量控制。

Abstract:

OBJECTIVE To develop an HPLC method with multiple wavelength ultraviolet for the simultaneous determination the content of 5-hydroxymethyl-2-furaldehyde, polygalaxanthone Ⅲ, atractylenolide Ⅰ, atractylenolide Ⅲ, salvianolic acid B, tanshinone ⅡA and catalpol in Mian’anning mixture. METHODS The sample was separated on a Hypersil C₁₈ column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of methanol-acetonitrile(1∶3) and 0.1% phosphoric acid solution. The flow rate was 0.7 mL·min^-1. The detection wavelength was set at 284 nm for 5-hydroxymethyl-2-furaldehyde, 320 nm for polygalaxanthone Ⅲ, 220 nm for atractylenolide Ⅰ and atractylenolide Ⅲ, 270 nm for salvianolic acid B and tanshinone ⅡA, 210 nm for catalpol. The column temperature was 25 ℃. RESULTS The calibration curves of 5-hydroxymethyl-2-furaldehyde, polygalaxanthone Ⅲ, atractylenolide Ⅰ, atractylenolide Ⅲ, salvianolic acid B, tanshinone ⅡA and catalpol were linear in the range of 10.08-100.80 μg·mL^-1(r=0.999 8), 3.55-35.50 μg·mL^-1(r=0.999 3), 5.80-58.00 μg·mL^-1(r=0.999 1), 4.20-42.00 μg·mL^-1(r=0.999 9), 40.50-405.00 μg·mL^-1(r=1.000), 12.28-122.80 μg·mL^-1(r=0.999 5), 7.90-79.00 μg·mL^-1(r=0.999 9), and the average recoveries(n=6) were 98.56%(RSD=0.97%), 96.94%(RSD=1.73%), 99.61% (RSD=1.03%), 97.54%(RSD=1.35%), 100.04%(RSD=0.86%), 98.59%(RSD=1.70%), 99.81%(RSD=1.43%), respectively. CONCLUSION The method is feasible, accurate and stable, which can control the quatity of Mian’anning Mixture.