【作者】 袁干军； 洪葵； 林海鹏；

【Author】 Yuan Ganjun1,2,3,Hong Kui1,2,*,Lin Haipeng2 (1.Hainan University,Haikou 570228; 2.Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101; 3.Hainan Medical College,Haikou 571101)

【机构】 海南大学； 中国热带农业科学院热带生物技术研究所； 海南医学院；

【摘要】 目的建立玫瑰孢链霉菌发酵液中达托霉素和A21978C组分的含量测定方法。方法以达托霉素为对照品,采用高效液相色谱法进行测定。色谱条件为色谱柱ShimadzuVP-ODS(250×4.6mm,5.0μm);梯度洗脱的流动相A:乙腈-0.05MpH7.2的磷酸缓冲液(30:70),流动相B:乙腈-0.05MpH7.2的磷酸缓冲液(70:30);流速1.0mL·min-1;检测波长为223nm,260nm,280nm,360nm。结果达托霉素和A21978C各组分之间完全分离,3个不同浓度的对照品溶液分别连续6次进样,峰面积的RSD≤0.3%;在0.55~22.00μg·mL-1的范围内,浓度(X)与峰面积(Y)具有良好的线性关系(r=0.9999),回归方程为:Y=0.4561X+0.0332;平均加样回收率为98.5%(RSD2.3%,n=9);检测限和定量限均为6.6ng(0.33μg·mL-1)。结论该方法灵敏准确,重现性好,适合于玫瑰孢链霉菌发酵液中达托霉素及A21978C组分的含量测定。更多还原

【Abstract】 Objective To develop a method for determination of daptomycin and A21978C components in fermentations produced by Streptomyces roseosporus.Methods Daptomycin, an A21978C component, is used as standard substance. A HPLC method was used, the system is equipped with a UV detector set at 223 nm, 260 nm, 280 nm, 360 nm and a Shimadzu VP-ODS column. a binary gradient elution was performed from mobile phase (A) acetonitrile-phosphate buffer (0.05 M, pH 7.2) (30:70, v/v) to mobile phase (B) acetonitrile-0.05 M phosphate buffer (0.05 M, pH 7.2) (50:50, v/v) at a flow rate of 1.0 mL·min-1.Results Daptomycin and A21978C components were complete separation each other, the RSD of peak area was not more than 0.3% with six sequences at three concentration levels;the calibration curve is linear at a concentration range of 0.55 to 22.00μg·mL-1(r = 0.9999), and the regression equation was Y =0.4 561 X+0.0 332; the average recovery was 98.5%(RSD 2.3%,n =9),and the limit of detection was 6.6 ng (0.33 μg·mL-1), which was equal to the limit of quantification.Conclusion The method is sensitive, accurate and good reappearance, and is propitious to the determination of daptomycin and A21978C components in fermentations produced by Streptomyces roseosporus NRRL 11379.更多还原