Anderson A D G, Poon P, Greenway G M, MacFie J
Combined Gastroenterology Research Group, Scarborough Hospital, Woodlands Drive, Scarborough YO12 6QL, UK.
Ann Clin Biochem. 2005 May;42(Pt 3):224-6. doi: 10.1258/0004563053857923.
Sucralose is a unique disaccharide probe which is stable in the colon and can be used to assess permeability over the whole gut. Additional information can be gained when sucralose is administered in combination with lactulose and a monosaccharide such as L-rhamnose in the form of a 'triple sugar test.' We describe a simple assay for urinary sucralose by HPLC with refractive index detection (HPLC-RI).
Phenyl-beta-D-glucopyranoside (internal standard) was added to 10 mL of urine, which was then passed through a 0.45 microm syringe filter. Elution was with 30% methanol (1 mL/min) on a reverse-phase C18 column. Detection was by refractive index, and integration based upon peak areas. Sixty standards of sucralose in human urine were analysed in order to quantify analytical variation.
The standard curve for urinary sucralose was linear from 25 to 500 mg/L (r>0.99). The limit of detection was 11 mg/L. Analytical recovery of sucralose at concentrations of 25, 50 and 100 mg/L was 101.5% (CV 7.59%), 102.9% (CV 5.82%) and 105.0% (CV 4.26%), respectively.
The technique described represents a simple assay for urinary sucralose which performed with acceptable accuracy and precision and should facilitate the use of the triple sugar test in clinical research.
三氯蔗糖是一种独特的二糖探针,在结肠中稳定,可用于评估全肠道的通透性。当三氯蔗糖与乳果糖和一种单糖(如L-鼠李糖)以“三糖试验”的形式联合使用时,可以获得更多信息。我们描述了一种通过高效液相色谱-示差折光检测法(HPLC-RI)测定尿中三氯蔗糖的简单方法。
将苯基-β-D-吡喃葡萄糖苷(内标)加入10 mL尿液中,然后通过0.45μm的注射器滤膜过滤。在反相C18柱上用30%甲醇(1 mL/min)洗脱。通过示差折光检测,并基于峰面积进行积分。分析了60份人尿中三氯蔗糖的标准品,以量化分析变异。
尿中三氯蔗糖的标准曲线在25至500 mg/L范围内呈线性(r>0.99)。检测限为11 mg/L。三氯蔗糖在25、50和100 mg/L浓度下的分析回收率分别为101.5%(CV 7.59%)、102.9%(CV 5.82%)和105.0%(CV 4.26%)。
所描述的技术代表了一种测定尿中三氯蔗糖的简单方法,其准确性和精密度可接受,应有助于在临床研究中使用三糖试验。