摘要:
目的 建立同时测定白念珠菌中4种多胺(腐胺、尸胺、亚精胺和精胺)的HPLC方法。 方法 样品经苯甲酰氯衍生化,资生堂C18色谱柱(100 mm×3.0 mm,3.0 μm)分离,以甲醇(A)-0.1%甲酸水(B)为流动相进行梯度洗脱,0~5 min,30%~48% B; 5~12 min,48% B;12~15 min,48%~70% B;15~25 min,70% B,流速0.6 ml/min,检测波长254 nm。 结果 腐胺、尸胺、亚精胺和精胺浓度分别在0.775~77.50 μg/ml(r=0.999 9)、1.160~116.0 μg/ml(r=0、999 9)、5.800~580.0 μg/ml(r=0.999 8)、2.533~192.7 μg/ml(r=0.999 9)的范围内线性关系良好。4种多胺的加样回收率为94.27%~109.3%,精密度RSD<4%。 结论 该含量测定方法灵敏、重现性好,能同时测定白念珠菌中4种多胺的含量。
Abstract:
Objective To develop a HPLC method for simultaneous determination of four polyamines (putrescine, cadaverine, spermidine, spermine) in Candida albicans. Methods HPLC analysis was performed on a Shiseido C18 column(100 mm×3.0 mm, 3.0 μm) after benzoyl chloride derivatization.The mobile phase consist of methanol (A) and water containing 0.1%methane acid (B). The gradient elution program was as follow:0~5 min, 30%~48% B; 5~12 min, 48% B;12~15 min, 48%~70% B;15~25 min, 70% B, the flow rate was 0.6 ml/min, the detection wavelength was 254 nm. Results The linearity was obtained over 0.775~77.50 μg/ml (r=0.999 9) for putrescine, 1.160~116.0 μg/ml (r=0.999 9) for cadaverine, 5.800~580.0 μg/ml (r=0.999 8) for spermidine and 2.533~192.7 μg/ml (r=0.999 9) for spermine. The average recoveries were between 94.27%~109.3%.The RSDs of precision were less than 4%. Conclusion The present study provided a sensitive and reproducible method for the simultaneously determination of four polyamines levels in Candida albicans.
点击查看大图
下载:
