Introduction: Ageratum conyzoides Linn is one of the Indonesian herbs are used as folk medicine. There are many studies to the therapeutic activity. However, the study of the influence in extraction parameters and how to standardize different batches of plant is still limited. Objective: The present study was designed to investigate the influence of extraction parameters on the variables matter, yield, phytochemical profile, and total quercetin from A. conyzoides L. leaves extract. The study also evaluates Liquid Chromatography-Mass Spectroscopy (LC-MS) and TLC fingerprint profiles of the extract to determine the proper quality of plant material of A. conyzoides L. leaves. Methods: Different temperatures (from 0°C to 60°C), solvent types (water and 70% ethanol (%v/v), and extraction time (192 h VS 2 h) were used to investigate the influence of extraction parameters. The profiles of extract were performed by TLC and LC-MS method. Results: The extract yield was influenced by temperature, solvent types, and time extraction. Our results suggest that the digestion technique at 60°C with 70% ethanol for 2h was recommended for the extraction method to obtained high yield and quercetin content as bioactive compound in A. conyzoides L. leaves extract. The range of quercetin content (40–52 ppm) combined with TLC and LC-MS profile can be used to determine the proper quality of plant material. Conclusion: All parameter of extraction have an affect on the yield extract and total quercetin on the final extract. Thus, there is a need an optimized method to obtain the suitable method.