Phellodendron amurense has been used for many years as a medical plant in traditional Chinese medicine and has shown great prospect in recent clinical trials for future applications. Berberine is an essential active compound contained in P. amurense. Our objective in this study was to quantify the content of berberine in P.amurense from sites at different elevations on Changbai Mountain. We collected samples of P. amurense from five different elevations on Changbai Mountain. Berberine in samples was extracted by ultrahigh pressure extraction(UPE). And the quantity was measured by high performance liquid chromatography(HPLC). First, the optimal HPLC conditions for berberine were identified with satisfactory precision(relative standard deviation, R SD5.6 %), good accuracy(relative error, R E 3.6 %) and good linear relation(R2= 0.9998) in the range of 6.576–328.8 mg L-1. Second, the combination of UPE and HPLC methods in quantitative analysis of berberine showed high repeatability(R SD= 3.28 %), reproducibility(R SD= 4.72 %),stability(R SD 1.27 %) and good recovery(99.54 %) for real plant materials. Samples from Heilongjiang Province at the lowest elevation contained the highest amount of berberine.Similarly, the lowest amount of berberine was recorded in samples from Changbai Forest Bureau of Jilin Province collected at the highest elevation in this paper. The proposed UPE–HPLC method is simple, reliable and low-cost for quantitative analysis of berberine. Content of berberine in P.amurense varied significantly by site on Changbai Mountain. Phellodendron amurense has been used for many years as a medical plant in traditional Chinese medicine and has shown great prospect in recent clinical trials for future applications. Berberine is an essential active compound contained in P. amurense. Our objective in this study was to quantify the content of berberine in P. elemese from sites at different elevations on Changbai Mountain. We collected samples of P. amurense from five different elevations on Changbai Mountain. Berberine in samples was extracted by ultrahigh pressure extraction (UPE). And the quantity was measured by First, the optimal HPLC conditions for berberine were identified with satisfactory precision (relative standard deviation, R SD 5.6%), good accuracy (relative error, RE 3.6%) and good linear relation (R2 = 0.9998) in the range of 6.576-328.8 mg L-1. Second, the combination of UPE and HPLC methods in quantitative analysis of berberine showed high repeatability (R SD = 3.28%), reproducibility (R SD = 4.72%), stability (R SD 1.27%) and good recovery (99.54%) for real plant materials. Samples from Heilongjiang Province at the lowest elevation contained the highest amount of berberine The similarity of the lowest amount of berberine was recorded in samples from Changbai Forest Bureau of Jilin Province collected at the highest elevation in this paper. The proposed UPE-HPLC method is simple, reliable and low-cost for quantitative analysis of berberine. Content of berberine in P.amurense varied significantly by site on Changbai Mountain.