Pemberton P W, Gagjee P, Chaloner C, Braganza J M, Lobley R W
University Department of Gastroenterology, Royal Infirmary, Manchester, England.
Clin Chim Acta. 1991 Jul 15;199(3):253-62. doi: 10.1016/0009-8981(91)90119-w.
Spectrofluorimetry was investigated as an alternative to HPLC for determining p-aminobenzoic acid and p-aminosalicylic acid in the N-benzoyl-L-tyrosyl-p-aminobenzoic acid/p-aminosalicylic acid test of pancreatic exocrine function. Urine specimens were hydrolysed for 30 min in 4 M NaOH at 100 degrees C. The fluorescence of p-aminobenzoic acid was measured in dimethyl sulphoxide solution (lambda ex = 300 nm, lambda em = 340 nm) and that of p-aminosalicylic acid in sodium acetate buffer, pH 4.0 (lambda ex = 297 nm, lambda em = 394 nm). The linear range was 0.038-8 mM for p-aminobenzoic acid and 0.051-12 mM for p-aminosalicylic acid, within-batch precision was 2.2% and 5.5%, respectively, and the entire analysis could be completed within 40 min. Although not eliminated, drug interference was greatly reduced in comparison with colorimetry. In 23 consecutive pancreatic function tests there was an excellent correlation between the p-aminobenzoic acid/p-aminosalicylic acid excretion index obtained by fluorimetry and the results from HPLC analysis (y = 0.914x + 0.070, r = 0.987, p less than 0.001). The method is simple, cost-effective and may be particularly valuable in developing countries having a high incidence of chronic pancreatitis.
在胰腺外分泌功能的N-苯甲酰-L-酪氨酸-p-氨基苯甲酸/p-氨基水杨酸试验中,研究了用荧光分光光度法替代高效液相色谱法来测定对氨基苯甲酸和对氨基水杨酸。尿标本在100℃下于4M氢氧化钠中水解30分钟。在二甲基亚砜溶液中(激发波长λex = 300nm,发射波长λem = 340nm)测定对氨基苯甲酸的荧光,在pH 4.0的醋酸钠缓冲液中(激发波长λex = 297nm,发射波长λem = 394nm)测定对氨基水杨酸的荧光。对氨基苯甲酸的线性范围为0.038 - 8mM,对氨基水杨酸的线性范围为0.051 - 12mM,批内精密度分别为2.2%和5.5%,整个分析可在40分钟内完成。与比色法相比,虽然药物干扰没有消除,但大大减少。在连续23次胰腺功能试验中,荧光法获得的对氨基苯甲酸/对氨基水杨酸排泄指数与高效液相色谱分析结果之间有极好的相关性(y = 0.914x + 0.070,r = 0.987,p < 0.001)。该方法简单、经济高效,在慢性胰腺炎发病率高的发展中国家可能特别有价值。