To explore the application of extraction and content determination in the prognosis of uncaria rhynchophylline and isorhynchophylline in Gambir Plant Nod. Methods: The uncaria rhynchophylline and isorhynchophylline extract was taken by water extract and methanol extract methods, and the content of the extract was determined by high performance liquid chromatography. Results: The good linear relationship of rhynchophylline was in the range of 2.5~80 μg/mL, the regression equation for the time Y=76.717 0X-0.072 7(r=0.999 8),RSD＜8.7%; In the linear relationship between the range of 2.0~80 μg/mL, rhynchophylline was good, the regression equation for the time Y=87.472 9X-0.366 6(r=0.999 7), RSD＜7.3%. The content of rhynchophylline and isorhynchophylline extracted by methanol extraction liquid was higher than those extracted by the water. And the best way to extract the contents was under the condition of 20 times the maximum amount of methanol, 24 hours of the invasion of cold time and 60 min of sonication. Conclusion: The methanol extraction method is prominent in extracting in uncaria rhynchophylline and isorhynchophylline, with good quality control, high repeatability, simple operation, and effect of improving prognosis, which is worthy of clinical application and promotion.