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Developing fast and sensitive methods to simultaneously analyze neurotransmitter and drug is of scientific significance for diagnosing and treating neurological diseases. Acupuncture can adjust and balance the blood gas, yin and yang and other physiological states through inserting acupuncture needle into the patient's body, so as to treat diseases, care health and strengthen body. In this study, the stainless-steel needle was used as electrode matrix for the sensor. Gold nanoparticles (Au NPs), polydopamine (PDA) and multi-wall carbon nanotubes (MWCNTs) were layer-by-layer modified onto the bare acupuncture needle (AN) through green and environment-friendly method, and the new interface showed efficient performance for simultaneous monitoring dopamine (DA), acetaminophen (AC) and chlorpromazine (CPZ). It was interesting that the synergistic interface of MWCNTs, PDA and AuNPs exhibits effect conductivity, and high electrocatalytic activity for three molecules. The sensor optimally exhibits wide linear ranges of 0.08-100 and 100-1000 mu M for DA, 0.07-100 and 100-1000 mu M for AC, 0.1-100 and 100-450 mu M for CPZ, respectively. The low detection limits are 0.015, 0.012, and 0.022 mu M correspondingly, which are competitively compared to previous reports. In addition, the sensor exhibits good anti-interference ability in the presence of excessive interference. The sensor can successfully monitor DA, AC and CPZ in human serum and urine with the recovery rates from 97.2 % to 102.6 %. The sensor showed good repeatability, stability, and reproducibility. The simultaneous detection of needlelike sensor should play important roles in biomedicine.