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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2.5~40μg·m L?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。
OBJECTIVE To establish a HPLC-ELSD method for the determination of peroxysorbendin in marine fungus Aspergillus sp. Methods The marine fungus Aspergillus sp. Was cultured by artificial fermentation. The fermentation broth was extracted with ethyl acetate and the macroporous resin was separated. The content of ergosterol in peroxisome was determined by HPLC-ELSD. A C18 column was used. The mobile phase consisted of methanol-water, gradient elution and evaporated light detector. The drift tube temperature was 75.0 ℃, the atomization temperature was 30.0 ℃, the carrier gas pressure was 40 psi and the gain was 40. Results The natural logarithm of injected quality had a good linear relationship with the natural logarithm of peak area in the range of 2.5-40 μg · m L · 1, with the average recovery of 100.1% and the precision test RSD of 1.9 %, Repeatability test RSD value of 1.61%, stability test RSD value of 1.46%. Conclusion The method has good separation effect, fast analysis speed and accurate results, and is suitable for the determination of peroxy-ergosterol in marine fungus Aspergillus sp.