1. Process Use of 3, 3 – Diaminobenzidine (DAB) as a brown stain in microscopy and immunohistochemistry. To reduce personnel exposure to Diaminobenzidine (DAB), it is recommended to use DAB Easytablets, Fisher catalog number: AC32800-5000; VWR catalog number: 200057-400 or Sigma catalog number:D3939. An alternative for DAB, which can be evaluated as a possible replacement, is Hanker Yates Reagent from PolySciences, Inc., catalog number: 08661. Hanker Yates Reagent produces a blue reaction product. Hanker Yates reagent is not mutagenic or a potential carcinogen. 2. Physical Hazards q Incompatibilities and Reactivities: Metals 3. Health Hazards q Eye: Causes eye irritation q Skin: Causes skin irritation q Ingestion: May cause irritation of the digestive tract. The toxicological properties of this substance have not been fully investigated. May cause nausea and vomiting. q Inhalation: Causes respiratory tract irritation. May cause irritation of the respiratory tract with burning pain in the nose and throat, coughing, wheezing, shortness of breath and pulmonary edema. q Target organ: Blood q Mutagen q Suspect Human Carcinogen 4. Personal Protective Equipment q EYE PROTECTION · Safety glasses, goggles or face shields shall be worn during operations in which DAB might contact the eyes (e.g., through splashes of solution). o Uvex Genesis Safety Glasses can be ordered from Fisher, catalog number: 19-040-143 or VWR, catalog number: 15001-568 · Ordinary (street) prescription glasses do not provide adequate protection. Adequate safety glasses must meet the requirements of the Practice for Occupational Education Eye and Face Protection (ANSI Z87.1-1989) and must be equipped with side shields. o Uvex Astro OTG, Fisher catalog number: 18-029 or VWR catalog number: 33000-506 · Contact lenses shall not be worn if the potential exists for DAB exposure. q HAND PROTECTION · Use of Kimberly-Clark Safeskin Purple Nitrile Exam Gloves are recommended for use with DAB. They are available through Fisher Scientific or VWR. q LAB COATS, ETC. · Lab coat, closed-toe shoes and long-sleeved clothing shall be worn when handling DAB. Protective clothing shall be worn to prevent any possibility of skin contact with DAB. q SAFETY SHOWER/EYEWASH · Where the eyes or body of any person may be exposed to DAB, suitable facilities for quick drench or flushing of the eyes and body shall be provided within the work area for immediate emergency use. · Bottle-type eyewash stations are not acceptable. 5. Special Handling Procedures and Storage Requirements q Handling: Avoid breathing dust, vapor, mist, or gas. Avoid contact with skin and eyes. 6. Waste Disposal Liquid Waste: Excess DAB and all waste material containing DAB must be placed in an unbreakable secondary container labeled with the following: "HAZARDOUS WASTE ". Full containers of DAB waste must be disposed of according to WFUHS EH&S Hazardous Waste Program. Solid Waste: See Decontamination of Equipment and Area
7. Decontamination of Equipment and Area Glassware: · Chlorine bleach is NOT effective in removing the mutagenic properties of DAB. · Potassium Permanganate Method The most commonly used one currently involves potassium permanganate and sulfuric acid. End products are known to be non-mutagenic. 1. Take up bulk quantities of diaminobenzidine tetrahydrochloride dehydrate in water and bulk quantities of the free base in 0.1 M hydrochloric acid so that the concentration of DAB does not exceed 0.9 mg/ml. Dilute solutions with the same buffer, if necessary, so that the concentration does not exceed 0.9 mg/ml. 2. For each 10 ml of solution, add 5 ml of 0.2 M potassium permanganate solution and 5 ml of 2 M sulfuric acid solution. 3. Allow the mixture to stand overnight, decolorize by the addition of sodium ascorbate, neutralize and dispose solution down the drain with copious amounts of water. · Contact WFUHS EH&S at 716-1221 for further decontamination information. 8. Spill and Accident Procedures In the event of a liquid spill, secure the area and contact WFUBMC Security at 716-9111. Reference The Safe Disposal of Diaminobenzidine. Lunn, George; Sansone, Eric B. Applied Occupational Environmental Hygiene, 6(1), January 1991. p.49-53. |