如意金黄散简化方3种有效成分的超声提取工艺优选

Optimization of extracting conditions for berberine,magnolol and emodin from simplified recipe of Ruyijinhuangsan

目的以小檗碱、厚朴酚和大黄素3种有效成分为指标,考察和筛选适于如意金黄散简化方新制剂制备工艺和质量控制研究的提取工艺。方法用HPLC法同时测定小檗碱、厚朴酚和大黄素,采用ACE.5C18-AR色谱柱,以甲醇-2%三乙胺(磷酸调pH 5.0)为流动相梯度洗脱,流速:1.0mL/min;柱温:30℃;双波长检测:λ1=294nm,λ2=345nm;进样量:10μL。以3种成分的提取率为指标,采用单因素试验和正交试验对其超声提取工艺进行优选。结果小檗碱、厚朴酚及大黄素的线性范围分别是:0.292～292mg/L(r2=0.999 8),0.292～292mg/L(r2=0.999 7),0.256～256mg/L(r2=0.999 5)。精密度RSD(n=6)分别为0.23%、0.17%、0.18%。供试品溶液12h内稳定性良好。平均回收率分别为98.26%(RSD 2.53%,n=6)、105.17%(RSD 2.06%,n=6)、100.39%(RSD 2.60%,n=6)。单因素试验考察了超声时间和次数、乙醇用量和浓度、酸浓度等因素,结果表明,随着超声时间增加、超声次数增多和乙醇含量提高,三组分的提取率均增加。结论正交试验结果表明最佳提取工艺为:95%乙醇(含1%盐酸)35mL,超声30min,提取3次。

Objective To establish a method for simultaneous determination of berberine, magnolol and emodin in the blend powder of Cortex Phellodendri Chinensis, Cortex Magnoliae Officinalis and Radix et Rhizoma Rhei, so as to provide evidence for preparation technique and quality control of simplified recipe of Ruyijinhuangsan. Methods High-performance liquid chromatography （HPLC） was used for simultaneous determination of berberine, magnolol and emodin at the following conditions： the column was ACE.5C18-AR, mobile phase was A： triethylamine, B： methanol. The flow rate was 1.0 mL/min, the temperature of column was 30℃, the detection wavelengths were set at 294 nm and 345 nm, and the injection volume was 10 μL. Using the yield ratio of three constituents as the assessment indices, we optimized the extraction technology by single factor test and orthogonal test L9 （34）. Results The calibration curves were linear within the range of 0.292-292 mg/L for berberine （r2=0.999 8）, 0.292-292 mg/L for magnolol （r2=0.999 7）, and 0.256-256 mg/L for emodin （r2=0.999 5）, with the average recovery rates being 98.26% （RSD 2.53%, n=6）, 105.17%（RSD 2.06%, n=6）, and 100.39% （RSD 2.60%, n=6）, respectively. The results of precisions were （n=6） 0.23%, 0.17%, 0.18%, respectively. The sample solutions were stable within 12 h. The single factor experiment analyzed ultrasonic duration and times, concentration of alcohol, the amount of solvent and the concentration of HCl. The results showed that, with the increase of ultrasonic duration, times and the alcohol concentration, the yield ratio of the three consitituents was increased.Conclusion The optimum options for extracting is as follows： ultrasonic extraction 3 times by 95% alcohol containing 0.1% HCl, with the volume at 35 mL, 30 min each time.