The preparation methods of heme include base matter-organic solvent method, glacial acetic acid method, acetone method, methanol or methanol-ethanol method, ion exchange cellulose method.
Method 1. Basic substance-organic solvent method
Add 60L of methanol and 1kg of diethylamine into the reaction tank, stir evenly, add 5kg of fresh bovine blood powder, stir and extract at 45°C for 1 hour, cool, filter, concentrate the filtrate to 5L, add 10L of glacial acetic acid, 200g of strontium chloride, and heat and distill Remove residual methanol, heat up to 100-102°C for 1 h, cool, filter, and collect hemin crystals. The hemin crystals were successively washed with glacial acetic acid, water and acetone, and dried to obtain 85 g of hemin.
Bovine blood meal [methanol, diethylamine]→[45℃, 1h][filter]→filtrate [ice Hac, SrCl2]→[100-102℃,1h][filter, wash, dry]→hemin
Take 60L of methanol and 5kg of spray-dried bovine erythrocytes into the reaction tank, stir evenly, extract at 40°C for 30 minutes, cool, filter, concentrate the filtrate to 5L, add 10L of glacial acetic acid, 100g of sodium chloride, add 2L of water to dissolve, heat and distill Remove residual methanol, heat up to 100°C for 1 h, cool, filter, and collect hemin crystals. The hemin crystals were washed successively with glacial acetic acid and water, and dried to obtain 65 g of hemin.
Bovine erythrocytes [methanol]→[40℃, 30min][filter]→filtrate [ice Hac, NaCl,H2O]→[100℃,1h][filter, wash, dry]→hemin
Preparation of heme: Take 300kg of fresh blood powder, add 1200L of methanol, add 30kg of dichloroacetic acid, stir well, stir and extract at 45°C for 1h, cool, filter, and concentrate the filtrate to dryness to obtain 45kg of heme.
Fresh blood meal [methanol, dichloroacetic acid] → [45°C, 1h] [filtered, concentrated] → heme
Preparation of hemin: Take 45 kg of hemin, add 100 L of glacial acetic acid containing 2 kg of NaCl, boil and reflux for extraction for 1 hour, cool and filter to obtain hemin, wash with glacial acetic acid and water in turn, and dry to obtain hemin 4.8kg.
Heme [mixed solvent of NaCl and glacial acetic acid]→[boiling for 1h][washing and drying]→hemin
Method two, methanol-ethanol mixed solvent extraction
Preparation of acidic blood meal: Take 100 kg of blood meal, add 100 kg of 0.6 mol/L HCl solution, stir for hemolysis, and spray dry to obtain 40 kg of acidic blood meal.
Blood meal [HCl] → acidic blood meal
Preparation of heme Take 40kg of acidic blood meal, extract 3 times with a mixed solvent composed of 90% ethanol, 6% methanol and 4% water, the amount of mixed solvent is 800L, 300L and 200L respectively, combine the supernatant, distill and concentrate to obtain heme.
Acid blood meal [mixed solvent] → [extract 3 times] supernatant [concentrate] → heme.
