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

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,5μm）;流速：0.7 m L·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10.08-100.80μg·mL^-1r=0.999 8）、3.55-35.50μg·mL^-1r=0.999 3）、5.80-58.00μg·mL^-1r=0.999 1）、4.20-42.00μg·mL^-1r=0.999 9）、40.50-405.00μg·mL^-1r=1.000）、12.28-122.80μg·mL^-1r=0.999 5）、7.90-79.00μg·mL^-1r=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。

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 λm） 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）, 3.55-35.50μg·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. KEY WORDS：