光密度法测定蛋白核小球藻生物量

Determination of Chlorella pyrenoidosa biomass using optical density method

目的修正传统光密度测定微藻生物量的方法。方法通过扫描并比较蛋白核小球藻藻液和总色素提取物在200～800nm范围内的吸光值,确定最佳波长以修正传统光密度法测定波长,绘制标准曲线来间接测定蛋白核小球藻的生物量。比较修正前后光密度法测定蛋白核小球藻生物量的精确性。结果 517nm下对应的吸光值与蛋白核小球藻的生物量有显著的线性关系。采用修正后光密度法测定干重量具有良好的线性关系:Y=0.257 8X-0.005 1(R2=0.995 6),相比传统光密度法具有更好的相关性;采用修正后光密度法测定细胞密度具有良好的线性关系:D=2 320.4X+0.030 6(R2=0.999 3),相比传统光密度法具有更好的相关性;修正后光密度法与干重法实时测定的蛋白核小球藻生物量结果基本一致,而传统光密度法与干重法具有一定的偏差。结论采用517nm修正传统光密度法测定波长,有效消除了色素干扰带来的误差,可以作为蛋白核小球藻生物量测定的特征波长。

Aim To rectify the traditional method of determining Chlorella pyrenoidosa biomass through optical density. Methods The absorbance of the Chlorella pyrenoidosa was scanned and compared and the total pigment from 200nm to 800nm as well, the properly specific wavelength identified and the standard curves generated to determine the biomass of ChloreUa pyrenoidosa, the accuracy of methods for determining Chlorella pyrenoidosa biomass through optical density is compared between the retified and the traditional. Results It has been found that the absorbance under 517nm correlated closely with the biomass of Chlorella pyrenoidosa. Measuring the dry weight through rectified optical density method reached significant linear relationship ： Y = 0. 257 8X - 0. 005 1 （ R^2 = 0. 995 6）, far more better than the traditional; measuring the cell density through rectified optical density method reached significant linear relationship ： D = 2 320. 4X ＋ 0. 030 6 （ R2 = 0. 999 3 ）, far more better than the traditional;the result of realtime determination for Chlorella pyrenoidosa biomass was basically in accordance between the rectified optical density method and the dry weight method, while different between, the traditional optical density method and the dry weight method. Conclusion The method of optical density is handy and rapid for the determi- nation of the biomass of Chlorella pyrenoidosa and eliminates the interference using the wave length of 517nm.