Waters Solution-Nucleotide Analysis in Dairy Products
Milk powder is added with various nucleotides such as uridine (UMP), cytidine (CMP), adenylate (AMP), guanylate (GMP), inosine (IMP), etc. Immunomodulatory function and memory.
Current challenges in nucleotide analysis:
Due to the high polarity of nucleotides, it is difficult to achieve good retention and separation with reversed-phase chromatography columns, so in order to improve the retention of nucleotides, ion-pair chromatography methods are often tried. Ion-pair chromatography methods have the following problems:
1. It is easy to foam, there are bubbles in the pipeline, which affects the analysis
2. Long balance time, prolonging analysis work time
3. Difficult to clean and damage the column
4. Easy to deteriorate, easy to block the pipeline and damage the instrument
Waters® solutions:
1. Avoid using ion pair reagents, only need to use volatile ammonium acetate, acetonitrile system
2. It is recommended to use Oasis® HLB SPE cartridges to prepare milk powder samples through purification to provide cleaner samples, improve sensitivity, and extend the life of the column and instrument
3. Use Amide chromatographic column analysis, and retain the polar compounds with a general reverse phase mobile phase, the method is simple and fast
Experimental results and chromatogram
Analysis results of 6-pin continuous injection of 5 kinds of nucleotide mixed standards
6-pin continuous injection analysis chart of actual sample
summary:
In this experiment, the Waters ACQUITY UPLC® H-Class system and BEHTM Amide 1.7μm 2.1 * 100mm chromatographic column were used to develop analytical methods for 5 nucleotides in infant milk powder. The experimental results show that:
1. On the Waters ACQUITY UPLC H-Class system, the BEH Amide 1.7μm 2.1 * 100mm column can be used to quickly separate 5 nucleotide standards in milk powder, and the resolution of the 5 compounds are all above 3.0; for the actual For milk powder samples, the separation between the five nucleotide samples and impurities is above 1.6.
2. The analysis method has good reproducibility. Among them, in the analysis results of 6 consecutive injections of milk powder samples, the RSD values ​​of the five kinds of nucleotide retention times were all less than 0.12%.
3. The Waters ACQUITY UPLC H-Class system has the Auto • Blend PlusTM function of the quaternary solvent. Therefore, method development on this system is very flexible and convenient, saves a lot of time for solvent configuration, and greatly improves the efficiency of the experiment. Thereby greatly reducing the solvent consumption of the experiment and reducing the cost of the experiment.
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