UPLC-MS/MS同时测定熟地黄中8个核苷类成分的含量

Simultaneous determination of eight nucleosides in Rehmanniae Radix Praeparata by UPLC-MS/MS

目的:建立超高效液相色谱-三重四极杆质谱法(UPLC-MS/MS法)同时测定熟地黄中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤8个核苷的含量。方法:采用ACQUITY UPLC BEH C_(18)(2.1 mm×50 mm,1.7μm)色谱柱,以0.1%甲酸溶液-甲醇为流动相梯度洗脱,流速0.2 mL·min^(-1),柱温30℃,进样量2μL,ESI+,多反应监测(MRM)对熟地黄饮片中8个核苷类成分进行含量测定。结果:所测8个核苷类成分在测定质量浓度范围内线性关系良好(R^2>0.996 0),平均加样回收率为98.6%～102.0%,RSD为1.3%～2.9%。6批样品中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤的含量范围分别为0.045～71.838、3.220～11.253、84.613～102.508、1.516～4.893、6.426～68.003、0.138～17.437、0.423～1.953、0.142～1.216μg·g^(-1)。结论:该方法适用于同时测定熟地黄饮片中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤8个核苷类成分含量,可用于熟地黄饮片的质量控制。

Objective:To establish an UPLC-MS/MS method for the simultaneous determination of guanosine,inosine,uridine,uracil,adenosine,cytidine,adenine and hypoxanthine in Rehmanniae Radix Praeparata.Methods:Waters BEH C18 column(2.1 mm×50 mm,1.7μm)with gradient elution of 0.1%formic acid and methanol was adopted.The flow rate was 0.2 mL·min-1 and the column temperature was 30℃.Eight nucleosides were detected in positive electrospray ionization mode and multiple reaction monitoring(MRM).Results:Eight nucleosides in Rehmanniae Radix Praeparata showed good linear relationships within their own ranges(R2>0.996 0),whose average recoveries were 98.6%-102.0%,with the RSDs of 1.3%-2.9%.The contents of eight nucleosides in 6 batches of samples were 0.045-71.838(guanosine),3.220-11.253(inosine),84.613-102.508(uridine),1.516-4.893(uracil),6.426-68.003(adenosine),0.138-17.437(cytidine),0.423-1.953(adenine)and 0.142-1.216(hypoxanthine)μg·g-1,respectively.Conclusion:The method for the simultaneous determination of the eight nucleosides can be used for qualitative and quantitative analysis of nucleosides in Rehmanniae Radix Praeparata.