應用 HPLC-ELSD 分析尿中葡萄糖含量的研究

中文摘要我們建立一個利用高效能液相層析法（high performance liquid chromatography, HPLC）搭配蒸發性光散射檢測器（evaporative light scattering detector, ELSD），以 maltose 為內部標準品，應用在尿中葡萄糖的分析方法。利用 PrevailTM Carbohydrate ES column（5 μm, 250 × 4.6 mm）作為分離，移動相為 acetonitrile : H2O = 80:20（v/v），流速為 1.2 mL/min，ELSD 之分析條件為：霧化器氣體流速 1.0 L/min，加熱管溫度 100 ℃。本方法在尿液葡萄糖之異日間分析精確度 1.98 % ~ 3.81 %。其準確度為 2.16 ~ 3.81 %。定量極限 limit of quantitation（LOQ）為 40 μg/mL。線性範圍為 40 ~ 480 μg/mL，其相關係數 R2 ≧ 0.9992 ± 0.0001。本方法並成功地運用在高雄市三民區 31位居民尿液檢品中葡萄糖含量的測定。

關鍵字

蒸發性光散射檢測器（ELSD）；高效能液相層析；葡萄糖

並列摘要

英文摘要 An analytical method for determination of glucose in urine has been developed. Glucose and maltose, the internal standard (I.S.), were analyzed by high performance liquid chromatography (HPLC) with evaporative light-scattering detector (ELSD). Separation was performed on a Prevail Carbohydrate ES column (5 μm, 250 mm × 4.6 mm) with acetonitrile : H2O at the rate of 80 : 20% (v/v) with flow rate of 1.2 mL/min. ELSD conditions were optimized at nebulizer-gas flow rate of 1.0 L/min and drift tube temperature of 100 ℃. For glucose in urine the inter-assay precision was 3.81% at 60 μg/mL, 2.76% at 120 μg/mL, 1.98% at 320 μg/mL. The relative errors (R.E.) and relative standard deviations (R.S.D.) were all ≦ 2.16 % and 3.81 %, respectively. The limit of quantitation (LOQ) was determined to 40 μg/mL. Linearity was found in the range of 40~480 μg/mL with a correlation coefficient of 0.9992 ± 0.0001. The mean value of slope and intercept were 0.0025 ± 0.0001 and 0.0093 ± 0.0037, respectively. The developed analytical procedure was successfully applied to determine glucose content in urine samples from 31 communities in Sanmin District of Kaohsiung city.