Objective: To establish an HPLC method for simultaneous determination of five nucleosides

uridine 5'-monophosphate

uridine

adenosine 5'-monophosphate monohydrate

guanosine

adenosine in Bailing capsules. Method: The separation was developed on an Ultimate AQ C18 column (4.6 mm×250 mm

5 μm) by gradient elution with methanol-0.05 mol·L-1 potassium dihydrogen phosphate solution

with flow rate at 1.0 mL·min-1 and column temperature at 25 ℃. The detection wavelength was at 260 nm. Result: The baseline separation of uridine 5'-monophosphate

uridine

adenosine 5'-monophosphate monohydrate

guanosine

and adenosine was successfully achieved. The results showed that these five ingredients had good linearity within the test ranges of 35.25-528.8(r=1.000)

38.07-571.1(r=1.000)

34.96-524.5(r=0.999 8)

38.34-575.1(r=1.000) and 39.84-597.6 ng(r=1.000)

respectively. The recovery were 101.2% (RSD 1.9%)

103.1% (RSD 1.1%)

96.7% (RSD 1.3%)

101.4% (RSD 2.7%) and 103.2% (RSD 1.6%)

respectively. Conclusion: The developed method of the HPLC-fingerprint and quantitative analysis showed good specificity

precision

reproducibility

stability and could be used for the quality control of Bailing capsules.