Chen Jing, Chen Jianhua, Zhou Changlin
School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.
J Chromatogr Sci. 2008 Nov-Dec;46(10):912-6. doi: 10.1093/chromsci/46.10.912.
High-performance liquid chromatographic (HPLC) methods were respectively developed for the quantitative determination of dihydroxyacetone (DHA) and glycerol in the fermentation broth. Validation parameters such as linearity, precision, accuracy, and specificity, limit of detection (LOD), and limit of quantitation (LOQ) were determined. Both HPLC methods were carried out on a Lichrospher 5-NH2 column with a mobile phase constituted of acetonitrile and water (90:10, v/v). The linearity range for DHA was 2.00-12.00 mg/mL with a correlation coefficient (r) of 0.9994. The LOD and LOQ were 0.06 and 1.20 mg/mL, respectively. The linearity range for glycerol was 0.50-20.00 mg/mL with a correlation coefficient of 0.9998. The LOD and LOQ were 0.22 and 0.50 mg/mL, respectively. Also, the HPLC method to determine DHA was compared with an existing visible spectrophotometric method. Statistical analysis by F-test and t-test showed no significant difference at 95% confidence level between the two methods when applied to low DHA concentrations while a large deviation existed in the determinations of high DHA concentrations. The HPLC method was more accurate to determine high DHA concentrations.
分别开发了高效液相色谱（HPLC）方法用于定量测定发酵液中的二羟基丙酮（DHA）和甘油。测定了线性、精密度、准确度、特异性、检测限（LOD）和定量限（LOQ）等验证参数。两种HPLC方法均在Lichrospher 5-NH2柱上进行，流动相由乙腈和水（90:10，v/v）组成。DHA的线性范围为2.00-12.00 mg/mL，相关系数（r）为0.9994。检测限和定量限分别为0.06和1.20 mg/mL。甘油的线性范围为0.50-20.00 mg/mL，相关系数为0.9998。检测限和定量限分别为0.22和0.50 mg/mL。此外，将测定DHA的HPLC方法与现有的可见分光光度法进行了比较。F检验和t检验的统计分析表明，当应用于低DHA浓度时，两种方法在95%置信水平下无显著差异，但在高DHA浓度测定中存在较大偏差。HPLC方法测定高DHA浓度时更准确。
