蜂王浆中磷酸腺苷的提取及超高效液相色谱分析

比较了高氯酸提取、热水提取和热硫酸镁溶液提取3种提取方式对蜂王浆中磷酸腺苷三磷酸腺苷(ATP)、二磷酸腺苷(ADP)和单磷酸腺苷(AMP)的提取效果,发现在低温(低于4 ℃)下以5%高氯酸的提取效果最佳。采用超高效液相色谱-紫外检测法分析蜂王浆中的ATP, ADP和AMP的含量。以BEH Shield RP18柱(100 mm×2.1 mm,1.7 μm)为分析柱,以50 mmol/L的磷酸二氢铵(pH 6.5)和乙腈为流动相进行梯度洗脱,3种磷酸腺苷在4 min内实现了较好的分离。以加标王浆样品作添加回收率测定,ATP, ADP和AMP的回收率分别为84.1%～94.3%,86.2%～93.7%和91.0%～104.3%,相对标准偏差均小于10%。方法已被用于一些实际样品的分析,以了解ATP, ADP和AMP在蜂王浆样品中的分布情况。

Extraction and ultra-performance liquid chromatographic analysis of adenosine phosphates in royal jelly

Several different extraction procedures including perchloric acid extraction, boiling water extraction and boiling magnesium sulfate solution extraction were studied for the extraction of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) from the royal jelly. Among these methods, the extraction with 5% perchloric acid at below 4 ℃ was the optimum extraction method. A simple, fast and sensitive ultra-performance liquid chromatographic (UPLC) method was developed for the determination of ATP, ADP and AMP in royal jelly. The separation was achieved within 4 min using a BEH Shield RP18 column (100 mm×2.1 mm, 1.7 μm) with 50 mmol/L monoammonium phosphate solution (pH 6.5) and acetonitrile as the mobile phase. The spiked recoveries of ATP, ADP and AMP were 84.1%-94.3%、86.2%-93.7% and 91.0%-104.3%, respectively. The relative standard deviations were less than 10%. This method was successfully applied to the analysis of some royal jelly samples from beekeepers and markets for the investigation of distribution of ATP, ADP and AMP in royal jelly samples.