WHOLE HUMAN BLOOD Blood isa body fluid in humans and animals that carries Essential constituents like nutri and allows the metabolic waste aways from cells, “Any person with good health can be a donor. Donor shouldn’t suffer from any transmitted disease. Haemoglobin content shoulda’ t less than 12.5g/dl of blood for female and 13.5g/dL of blood for male, Blood is around 7% of whole body weight with total volume of 5-7 L of which comprises of plasma and different kinds of cells. Whole blood is combination of cellular components, colloids and crystalloids. Collection- + Blood is collected aseptically from median cubital vein. im front of the elbow into a sterile container containing anti coagulant. + During collection the container is gently shaken, to cnsure that blood and anti coagulate are mixed well which. ‘prevents the formation of Fibrin clots. + Atatime not more than 420ml blood is taken. ‘The collected blood is sealed and freezes at 4-6°C Blod clotting factors: PROTHROMBIN _Trmoymor ASN AND CALCHUNHIONS THREEWHOLE HUMAN BLOOD + Heparin- naturally occurring anti-coagulant, expensive and looses its anticoagulant activity in in-vitro condition. + EDTA Di sodium salt- chelating agent and prevents blood coagulation but less stable, + Gitrates-Acid citrate dextrose(ACD) solution is the most commonly used Anti coagulant, Processing or testing after collection: = Rh system: it consist of 50 different types of blood group antigens also determine that whether doner’s blood is: having Rhesus factor or not ie. Rh+/Rh- + Rh factor- group of antigen in RBCs. This antigen will cause agglutination when it reacts with antibodies from Individuals without this antigen. + Determination of ABO grouping and determination of transmitted disease ‘Table 1.4 Blood grouping Storage: ae el * Sterile comtainer/ disposable bags containing anticoagulants, + Actemperature 4-6°G until required for use, Can be stored for 42days. + Ifstored ant room temperature (for I day), it may reduce post transfusion survival of Erythrocytes.DUCTS: COLLECTION, PROCESS DRIED HU + Insome circumstances . dried human plasma is used as substitute for whole blood. + The majoradvantage is . iit is stored properly can be used upto 5 years, ifprotected from light can be keptat 20° from refrigerator, can be given to patients of any blood group. Collection: = ‘The whole human citrated blood is used as source and supernatant fluid Is separated by centrifugation. * Batches of not more than 10 bottles are collected , mixed in correct ratio to neutralise Agglutinins (plasma antibody). + The mixture is stored at 4-6 °C (sterility testing is also done periodically). Processing: Preliminary freezing- 400 mL mixture is taken in the boutes and sealed with bacteriologically efficient pads and centrifuged at -18°C, The liquid snap-freezes and become distressed inside the boule. Primary drying: the bottles of frozen material are mounted horizontally in the drying: chamber under high vacuum for 2 days after which the moisture content of residual is about 2%, Secondary drying. then the boule are transferred to another chamber by vacuum desiccation over phosphorus. ‘peroxide. For one day product is left with only 0.5% moisture content.DRIED TIUMAN PLASMA Dircetion of use: Dried plasma should be reconstituted in Water for injection Sodium chloride injection Solution containing 2.5% dextrose and 0.45% sodium chloride. Must be used immediately after reconstitution. * Dried plasma must be soluble in these solvents and form solution within 10min, * fel formation wok place / incomplete solution is obtained . ILindicates dried plasma is deteriorated. Uses: + Mostly preferred in the treatment of severe burns where excessive loss of fluid and protein oceurs. "> May be given as substitutes of whole human blood (condition-in emergency whole Human blood is not available/ blood group matching; tests are not known)‘These are of non human origin , which could be use to restore the blood volume temporary. Properties ofsuch substitutes are: ‘Should have same osmotic pressure as whole blood. ‘Should have same viscosity as plasma. ‘Should have low rate of excretion but must completely eliminate form the body. Free from antigen, pyrogen and must be sterile ‘Stable in liquid form at normal and sterilising vemperanure, Economic and easy to prepare. "+ Examples of Plasma substitutes: 1. Gum saline (injection of sodium chloride and Acacia) British Pharmacopeia in 1932 prepared Gum saline by mixing: 6% Acacia in 0.9% NaCl solution. 2. PVP- (Polyvinylpyrrolidone) in 1950's century it was used a plasma substitute but has major problem was its Careinogenie nature. ‘Dextran- Ibis the most satisfactory plasma substitute.it was discovered by LOUIS PASTEUR as microbial product inPLASMA SUBTITUTES bi * Inoculum - Leuconostoc mesenteroides. This organism secrets an enzyme Dextran sucrase which converts sucrose into dextran. + Medium composition - Sucrose, peptones, yeast extract, Dipotassium hydrogen phosphate, manganese chloride and calcium chloride, © Prepration- Fresh culture of Z.mesenreroides is added to fermentation medium, Fixed the fermentation at 80 °G and incubate for 20 hours at pH 6-8 ¥ fermentation broth isadded with equal volume of chilled ethanol for precipitation ofdextran Centrifuge at 10,000rpm for 15min Discarded Supernatant Precipitates are dried under in vacuum « in presence of Calcium chloride: ‘The dextran produced may have high molecular weight which cannot be used as plasma substitutes as it may yield very viscous solution which may not be administered parenterally. It may cause renal damage and allergic reaction. So molecular weight of Dextran ean be reduced by following three methods Le.Acid hydrolysis, Thermal Degradation and Ultra sonication, Flowchart L6 Production ofDesiranPLASMA SUBTITUTES ‘Three methods of molecular weight reduction of dextran, ¥ + + eS “Therinal Degradation Ula Soalcation ope er Dsate acluciet restate ee eraetoaren Bioensa Saicentions tuted w2 a 90°C. Th cease coran pene en ems pieced Eee erence rer eeeied aan ere ee erties comttara eater areas ee eater ere is 10,0000(1 lakh) to 000 (2k thousand) daltons. Quality control tows performed Pyrogen testing. testing andl ysical testing for determination of antigen. Example: injections like Dextran 110 and Dextran 40. " Uses- in treatment of severe burns and in treatment ofacute Peritonitis.