Peroxidase exists in different amount in herbal sources. Peroxidase is a glycoprotein which contains heme and includes different cation and anion isoenzymes that categorized in 3 groups: acid (pI<4), neuter (4<pI<10) and alkali (pI> 11). This enzyme is resistant against variable pH and has high thermal stability and has many applications in laboratory diagnosis for measurement hormones and bacterial toxins with immunoassay method in ELISA technique. In this study, we isolate and purify different peroxidase isoenzymes from Raphanus sativus l.niger. The crude enzyme was extracted from raw root plant by homogenization and concentrated crude juice using ammonium sulfate precipitation, the active fractions were collected by assaying peroxidase special method, three isoenzymes observed in primary gel electrophoresis then with purification and apply ion exchange chromatography include Q and SP- Sepharose, we observed two peroxidase isoenzymes. Isolation process of these two isoenzymes was done using SP-Sepharose chromatography procedure. In addition, we examine the detection of hydroxy aromatic derivations (phenol and benzidin) using entrapment of the peroxidase in polyacrylamide gel.Using a solution contains sodium acetate buffer and hydrogen peroxide, a peace slide of entrapment enzyme in gel in, can clearly detect phenol and benzidin in very short time. Therefore, we suggest using this model; ones could easily eliminate phenol and benzidin as primary pollution of much industrial sewage. We hope that by using abundant and cheap sources, enzyme production in our country will progress and be cost effective and we will be able to eliminate our industrial pollutions.

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