HPLC-ELSD测定海洋真菌Aspergillus sp.中过氧麦角甾醇的含量

Determination of Ergosterol Peroxide in Marine Fungi Aspergillus sp. by HPLC-ELSD

关键词:  高效液相色谱法  海洋真菌  过氧麦角甾醇

Key words:  HPLC  marine fungi  ergosterol peroxide

摘要:

目的 建立高效液相色谱-蒸发光散射法（HPLC-ELSD）测定海洋真菌Aspergillus sp.中过氧麦角甾醇含量的方法。方法 人工发酵培养海洋真菌Aspergillus sp.，发酵液经乙酸乙酯萃取，大孔树脂分离，用HPLC-ELSD测定过氧麦角甾醇含量。采用C₁₈色谱柱，流动相为甲醇-水，梯度洗脱，蒸发光检测器：漂移管温度为75.0℃，雾化温度为30.0℃，载气压力为40 psi，增益值为40。结果 过氧麦角甾醇在2.5~40 μg·mL^-1内，进样质量的自然对数值与峰面积的自然对数值线性关系良好，平均加样回收率为100.1%，精密度试验RSD值为1.9%，重复性试验RSD值为1.61%，稳定性试验RSD值为1.46%。结论 本方法分离效果良好，分析速度快，结果准确，适用于海洋真菌Aspergillus sp.中过氧麦角甾醇的含量的测定。

Abstract:

OBJECTIVE To establish the method for determining ergosterol peroxide in marine fungi Aspergillus sp. by HPLC-ELSD. METHODS Marine fungi Aspergillus sp. was cultured by artificial fermentation, fermentation broth was extracted by ethyl acetate, and determined by HPLC-ELSD. C₁₈ column was used with the mobile phase of methanol and water gradient elution, and the determination condition of evaporative light detector was drift tube temperature of 75.0℃, atomization temperature of 30.0℃, carrier gas pressure of 40 psi, the gain was 40. RESULTS Pair value of sample quality and peak area had a good numerical linear relationship in the ergosterol peroxide concentration range of 2.5-40 μg·mL^-1, the average recovery was 100.1%, the RSD value of precision test was 1.9%, the RSD value of repeatability test was 1.61%, the RSD value of stability test was 1.46%. CONCLUSION The method has a good separation, quick analysis speed and accurate result, it is suitable for the determination of marine fungus Aspergillus sp. of peroxy ergosterol content.