Lassek E, Montag A
Z Lebensm Unters Forsch. 1987 May;184(5):361-5. doi: 10.1007/BF01126658.
The following method was developed for the qualitative and quantitative determination of purine and pyrimidine bases in carbohydrate rich food: The bases were liberated from nucleic acids, nucleotides or nucleosides by acid hydrolysis in a pressure digestion vessel. A complete liberation without losses of purine bases occurs upon hydrolysis for 15 min at 240 degrees C with trifluoroacetic and formic acids (1+1; V + V), pyrimidine bases need 45 min at 240 degrees C. The products arising from side reactions (such as hydroxymethylfurfural from hexoses and furfural from pentoses) could be removed from the hydrolysate by extraction with dichlormethane. The liberated bases could be separated upon stepwise elution by cation exchange chromatography. They were detected and determined by UV-measurements, continuously monitoring at lambda = 260 nm, and integrating electronically. The evaluation was carried out by a method with internal standard.
在压力消解容器中通过酸水解从核酸、核苷酸或核苷中释放出碱基。在240℃下用三氟乙酸和甲酸(1 + 1;V + V)水解15分钟可使嘌呤碱基完全释放且无损失,嘧啶碱基在240℃下需要45分钟。副反应产生的产物(如己糖产生的羟甲基糠醛和戊糖产生的糠醛)可通过用二氯甲烷萃取从水解产物中除去。释放出的碱基可通过阳离子交换色谱法进行分步洗脱分离。通过紫外测量进行检测和定量,在λ = 260nm处连续监测并进行电子积分。通过内标法进行评估。