UPLC-MS/MS同时测定熟地黄中8个核苷类成分的含量

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 C₁₈（2.1 mm×50 mm，1.7 μm）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2 μL，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（R²>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、0.142~1.216 μg·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, 1.7 μm) 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.

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