高效液相色谱法检测胃液中色氨酸和利多卡因

Investigation of Tryptophan in Gastric Juice by High Performance Liquid Chromatography Coupling with Ultraviolet,Fluorescence Detection and Mass Spectrometric Identification

采用高效液相色谱-紫外-荧光和高效液相色谱-紫外-质谱联用法,证明胃癌患者胃液中存在的荧光物质主要为色氨酸。建立了反相高效液相色谱-双波长同时紫外检测法定量分析胃液中色氨酸和利多卡因的方法。使用Kromasil C18柱(150mm×4.6mm,5μm),以含0.1%三氟乙酸的甲醇溶液与含0.1%三氟乙酸水溶液为流动相,梯度洗脱分析。色氨酸使用278nm检测,浓度在0.5～200mg/L范围内峰面积对浓度的线性相关系数为0.9994;利多卡因使用254nm检测,浓度在20～5000mg/L范围内相关系数为0.9992。色氨酸和利多卡因的检出限分别为0.15和5mg/L;平均加标回收率分别为70.8%～110.4%(色氨酸)和87.1%～116.2%(利多卡因)。利用本方法对38例胃癌患者和48例非胃癌患者的胃液进行测试,两组胃液中色氨酸的含量存在一定的差异。

The HPLC-UV-Fluorescence technique and HPLC-UV-MS technique were used to determine fluorescence molecule in gastric juice of patients with malignant gastric cancer, which is identified as tryptophan. Lidocaine was detected in gastric juice of patients who were examined by gastric endoscopy. A method was developed to determine concentration of tryptophan and lidocaine in gastric juice using HPLC-UV technique, in which Kromasil C18 column（ 150 mm ×4.6 mm, 5 μm） and methanol（ containing 0.1% trifluoroacetic acid （TFA））-water（ containing 0.1% TFA） as gradient mobile phase were used. The linear relationships were obtained between the concentration of tryptophan in the range of 0.5 - 200 mg/L and its peak area at 278 nm with correlation coefficient of 0.9994, and between the concentration of lidocaine in the range of 20 - 5000 mg/L and its peak area at 254 nm with correlation coefficient of 0.9992. The limit of detection is 0.15 mg/L for tryptophan and 5 mg/L for lidocaine. The recovery is 70.8% - 110.4% for tryptophan and 87.1% - 116.2% for lidoeaine. 38 gastric juice samples of patients with malignant gastric disease and 48 gastric juice samples of patients with benign gastric disease were tested and the concentration of tryptophan in those two groups of gastric iuice samples was aoparentlv different.