AbstractThis study established a spectrum–effect relationship method for screening and quantifying the analgesic and anti‐inflammatory active ingredients in Angelicae Pubescentis Radix (AP) by ultra‐high‐performance liquid chromatography–quadrupole mass spectrometry detector analysis (UPLC–QDA). First, the fingerprint of AP was established to determine the common peaks. Next, six batches of AP samples, with significant differences, were selected for evaluation of pharmacological activity. Subsequently, the spectrum–effect relationship was used to screen the active ingredients. Finally, the screened ingredients were quantified using UPLC–QDA. In total, 21 common peaks were identified and four effective compounds (bergapten, columbianetin acetate, osthole and isoimperatorin) were selected using the gray relational analysis and partial least squares regression analysis. Quantitative analysis showed that the content of the four effective compounds was the highest in a randomly selected batch, S7 (Hubei). To our knowledge, this is the first attempt that evaluated the quality and spectrum–effect relationship of AP by quantitative analysis and chemometrics. This study identified the key pharmacologically active components of AP and thereby improved the quality evaluation system of AP. This method has broad application prospects for screening effective components and will be helpful in establishing more reliable, scientific and reasonable quality standards for AP and other traditional Chinese medicines.