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目的:建立一种同时测定贝母药材中核5种苷类化合物尿苷、腺嘌呤、鸟苷、胸苷、腺苷的高效液相色谱法。方法:采用Welch Materials XB-C18色谱柱(4.6 mm×250 mm,5μm),甲醇-5 mmol.L-1乙酸铵-乙酸缓冲盐(pH4.30)作为流动相,以1 mL.min-1的流速进行梯度洗脱(洗脱梯度程序为:0~10 min,0%~1%A;10~20 min,1%~5%A;20~25 min,5%A;25~35 min,5%~30%A;35~37 min,30%~0%A;37~40 min,0%A);进样量20μL;检测波长260 nm。结果:在0.24~13.60 mg.L-1,各核苷类化合物的响应峰面积与其相应的浓度呈现良好的线性关系,r>0.998 3;各待测物的日内精密度和日间精密度的RSD均小于2.1%;重复性良好(RSD<5.5%);回收率范围在93.55%~101.88%,RSD<3.0%。结论:不同种贝母药材中核苷类化合物的含量大致顺序为湖北贝母>浙贝>川贝≈平贝;本方法简便、可靠、准确,可用于贝母药材中核苷类化合物的含量测定,为全面开发利用贝母药材提供进一步依据。
OBJECTIVE: To establish a HPLC method for simultaneous determination of uridine, adenine, guanosine, thymidine and adenosine in five kinds of glycosides in Fritillaria. METHODS: The samples were separated on a Welch Materials XB-C18 column (4.6 mm × 250 mm, 5 μm) and methanol-5 mmol·L -1 ammonium acetate-acetate buffer (pH 4.30) (Gradient elution procedure: 0-10 min, 0% -1% A; 10-20 min, 1-5% A; 20-25 min, 5% A; 25-35 min , 5% -30% A; 35-37 min, 30% -0% A; 37-40 min, 0% A); injection volume 20μL; detection wavelength 260 nm. Results: In the range of 0.24 ~ 13.60 mg.L-1, the peak area of each nucleoside compound showed a good linear relationship with its corresponding concentration, r> 0.998 3. The intra- and inter-day precision of each analyte RSD less than 2.1%, good reproducibility (RSD <5.5%), recoveries ranged from 93.55% to 101.88% and RSD <3.0%. Conclusion: The content of nucleosides in different species of Fritillaria is roughly in the order of Fritillaria> Fritillaria> Fritillaria cirrhosa × Fritillaria; the method is simple, reliable and accurate, which can be used to determine the content of nucleosides in Fritillaria, For further development and utilization of Fritillaria provide further basis.