Objective: To establish an HPLC method for simultaneous determination of eight nucleosides in Rhizoma Arisaema

Rhizoma Pinellie and Rhizoma Typhonii. Method: The chromatographic separation was achieved on a Phenomenex Luna PFP (2) (4.6 mm×250 mm

5 μm) column with methanol-water solution as mobile phase (gradient elution). The flow rate was 0.8 mL·min-1

the column temperature was maintained at 35℃ and the detective wavelength was set at 260 nm. Result: The linear ranges of uracil

hypoxanthine

xanthine

uridine

inosine

guanosine

thymidine and adenosine were 0.004 34-0.434 μg (r=0.999 9)

0.002 34-0.234 μg (r=0.999 9)

0.004 07-0.407 μg (r=0.999 7)

0.010 90-1.090 μg (r=0.999 9)

0.002 01-0.201 μg (r=0.999 9)

0.007 32-0.732 μg (r=0.999 9)

0.001 22-0.122 μg (r=0.999 7)

0.002 07-0.207 μg (r=0.999 9) respectively. The average recoveries of the eight nucleosides were 104.2%

98.7%

103.7%

99.4%

101.8%

102.9%

98.3%

102.3% and RSDs were 2.1%

2.4%

3.0%

1.4%

2.0%

1.9%

2.8%

2.7% respectively. Conclusion: The method is simple

accurate and rapid. It can be used for determination of nucleosides in Rhizoma Arisaema

Rhizoma Pinellie and Rhizoma Typhonii.