A novel and highly specific bulk membrane optode was developed for the ultra-sensitive detection of zinc(II) in biological, pharmaceutical, and water samples. The polymer inclusion membrane (PIM) utilized in this study consists of 50% polyvinyl chloride (PVC) as a base polymer, 9.00% DOP (dioctylphthalate) as the plasticizer, and 40.0% D2EHPA (di(2-ethlyhexyl)phosphoric acid) as the carrier. To facilitate the spectrophotometric determination of zinc(II), a colorimetric reagent, namely 4-(2-arsonophenylazo) salicylic acid (APASA) {1.00%, m/v}, was employed. When Zn(II) was extracted into the PIM, it led to the creation of the zinc-D2EHPA complex. This complex then underwent a reaction with APASA, resulting in the formation of a red Zn−APASA complex with a maximum absorption wavelength (λmax) of 558 nm. To optimize the response of the optode, a central composite design was employed, considering variables such as the amount of additive and reagent, response time, and pH. When operated under the specific optimal conditions, the sensor demonstrated a limit of quantification (LOQ) of 0.74 ng/mL (equivalent to 1.17 × 10−8 M) and a limit of detection (LOD) of 0.22 ng/mL (equivalent to 3.44 × 10−9 M). The optode membrane demonstrated excellent reproducibility, stability, and a relatively long lifespan, making it suitable for precise and accurate monitoring of Zn(II) ion content. Regeneration of the optode was achieved effectively using 0.25 nitric acid solution, and its response exhibited reversibility and reproducibility, showed a relative standard deviation of less than 1.33%. Moreover, the PIM-APASA optode exhibited a high level of effectiveness in accurately determining the presence of Zn(II) ions in real environmental samples.