Lababpour Abdolmajid, Lee Choul-Gyun
Department of Biotechnology, Institute of Industrial Biotechnology, Inha University, 253 Younghyun-Dong, Nam-Gu, Incheon 402-751, Republic of Korea.
J Biosci Bioeng. 2006 Feb;101(2):104-10. doi: 10.1263/jbb.101.104.
A first-order derivative spectrophotometric method has been developed for the simultaneous measurement of chlorophyll and astaxanthin concentrations in Haematococcus pluvialis cells. Acetone was selected for the extraction of pigments because of its good sensitivity and low toxicity compared with other organic solvents tested; the tested solvents included acetone, methanol, hexane, chloroform, n-propanol, and acetonitrile. A first-order derivative spectrophotometric method was used to eliminate the effects of the overlaping of the chlorophyll and astaxanthin peaks. The linear ranges in 1D evaluation were from 0.50 to 20.0 microg x ml(-1) for chlorophyll and from 1.00 to 12.0 microg x ml(-1) for astaxanthin. The limits of detection of the analytical procedure were found to be 0.35 microg x ml(-1) for chlorophyll and 0.25 microg x ml(-1) for astaxanthin. The relative standard deviations for the determination of 7.0 microg x ml(-1) chlorophyll and 5.0 microg x ml(-1) astaxanthin were 1.2% and 1.1%, respectively. The procedure was found to be simple, rapid, and reliable. This method was successfully applied to the determination of chlorophyll and astaxanthin concentrations in H. pluvialis cells. A good agreement was achieved between the results obtained by the proposed method and HPLC method.
已开发出一种一阶导数分光光度法，用于同时测定雨生红球藻细胞中叶绿素和虾青素的浓度。与其他测试的有机溶剂（包括丙酮、甲醇、己烷、氯仿、正丙醇和乙腈）相比，丙酮因具有良好的灵敏度和低毒性而被选作色素提取剂。采用一阶导数分光光度法消除叶绿素和虾青素峰重叠的影响。一维评估中的线性范围为：叶绿素0.50至20.0μg·ml⁻¹，虾青素1.00至12.0μg·ml⁻¹。该分析方法的检测限为：叶绿素0.35μg·ml⁻¹，虾青素0.25μg·ml⁻¹。测定7.0μg·ml⁻¹叶绿素和5.0μg·ml⁻¹虾青素的相对标准偏差分别为1.2%和1.1%。该方法简便、快速且可靠。该方法已成功应用于雨生红球藻细胞中叶绿素和虾青素浓度的测定。所提方法与高效液相色谱法所得结果之间具有良好的一致性。
