Heparin solution preparation

Figure 4 shows the relatively even adsorption on polyvinylpyridine surfaces spread by knife coating and dipped into radioactive heparin solution (500 fig./ml.) for 6 minutes. Six discs each 1 inch in diameter were die cut from each of two duplicate preparations which were similar in visual evenness to the ones shown. Planchet counts gave the results shown below ... 

The ability of heparin to complex ions and simple bases is important in various pharmacological preparations. The decontraction or spasmolytic action of the isolated atropinized guinea-pig ileum after histamine contraction, however, is due to phenol and cresol in heparin solutions and not to heparin itself . Green, Day and Roberts draw attention to the fact that heparin preparations contain more amino compounds than can be accounted for by the glucosamine , and it is these which cause the contraction of guinea-pig ileum. The contraction is different in shape from that of histamine, is not prevented by antihistamines, and is similar to that of polypeptides. Heparin also inhibits trypsin release of bradykinin from serum . 

Heparin and heparinoids are absorbed in only trace amounts when given in large doses orally . EDTA increases the absorption . Subcutaneous and intramuscular injection of various depot heparin preparations have not been found very satisfactory. They fail to give satisfactory blood levels and they increase Sensitization. With subcutaneous injection, the ability of heparin to become fixed to protein becomes a factor modifying absorption. One of the basic difficulties in deciding on the value of subcutaneous or intramuscular injections of heparin is that there has been no quantitative comparison made of the blood levels of heparin administered in these two ways, to determine how much appears in the circulation in active form. It is possible that much heparin never reaches the circulation. This is to be expected when it is remembered that heparin released by mast cells does not reach the circulation. Depository forms of heparin have been replaced by the use of concentrated heparin solutions (40 gm per cent), so that volumes of 0-2 to 0-4 ml. ean be injected into subcutaneous fat tissue or intramuscularly. 

Isolation and Characterization of Heparin Fractions. Preparative scale fractionation was carried out by sequential extraction of eight tubes, each containing 10 mg of heparin in 20 mL of 1-butanol (upper phase) and 20 ml of 0.01 M NaCl (lower phase). The biphasic solution contained 30 g HPC/ of butanol phase. After removing the lower, aqueous phase, the upper butanol phases of each tube were successively extracted with 4 mL of aqueous butanol-saturated solutions of NaCl containing 2 g/L HPC as listed in Table I. Fractions are identified by the NaCl concentration at which the fraction was extracted. Equivalent fractions from different tubes were combined, and, after making each tube at least 0.5 M in NaCl, the heparin was precipitated with three volumes of acetone. A more detailed description of the procedure was published previously (11). Analytical-scale fractionation of the three affinity-fractionated heparins was carried out similarly, except that only 1 v ol (as uronate) of each was fractionated. Molecular weight distributions of the fractions were determined on 6% agarose as described (11). 

Heparin solution, 1 ml Prepare 10 mg/ml solution of Sigma grade 1-A sodium salt (H3393) in 50% glycerol/ddH20 store at -20°C. 

By filling the lumen of the catheter with such a solution the formation of blood clots is prevented and the flow is maintained. Sodium citrate solution is an alternative for a concentrated heparin solution and should be preferred because of the anti-microbiological effect [20]. Citra-Lock is available as a medical device. This product contains 46,7 % sodium citrate and is CE registered class lib. Are there justifiable reasons to prepare the solution This could be for example when the marketed product is associated with more side effects due to the higher concentration, or is delivered in a container that is hard to use in practice. If those reasons are absent, then the marketed product is to be preferred. 

Heparin was prepared in 5 mM sodium phosphate buffer (pH 7.0) containing 150 mM sodium chloride at 10 mg/mL. To 80 pi of heparin, 20 pi of heparin lyase (0.03 lU in the same buffer) was added. The reaction mixture was incubated overnight at 30 C. After the reaction was complete, protein was removed from the reaction mixture by adjusting its pH to 3.0 and passing this solution through a 1 mL column of sulfopropyl-Sephadex adjusted with 30 mM hydrochloric acid to the same pH. The material fail-... 

The analysis of human plasma for acetaminophen, the active ingredient in some pain relievers, involves a unique extraction procedure. Small-volume samples (approximately 200 fiL) of heparinized plasma, which is plasma that is treated with heparin, a natural anticoagulant found in biological tissue, are first placed in centrifuge tubes and treated with 1 N HC1 to adjust the pH. Ethyl acetate is then added to extract the acetaminophen from the samples. The tubes are vortexed, and after allowed to separate, the ethyl acetate layer containing the analyte is decanted. The resulting solutions are evaporated to dryness and then reconstituted with an 18% methanol solution, which is the final sample preparation step before HPLC analysis. The procedure is a challenge because the initial sample size is so small. 

The cholinesterase-inhibiting activity of the phosphorofluoridates was compared quantitatively with that of eserine sulphate thus. To 0-2 ml. of heparinized human plasma was added 05 ml. of a solution containing either eserine or the phosphorofluoridate in varying concentrations then the mixture was kept at room temperature for 10 min. before 1 /tg. of acetylcholine in 1 c.c. saline solution was added. After 5 min. at room temperature, the mixture was made up to 10 ml. with frog saline containing eserine 1/100,000, which at once stopped the action of any cholinesterase not yet inactivated. The solution was then assayed for acetylcholine on the frog rectus-muscle preparation. 

Heparin (from pig intestinal mucosa). Most likely contaminants are mucopolysaccharides including heparin sulphate and dermatan sulphate. Purified by pptn with cetylpyridinium chloride from saturated solutions of high ionic strength. [Cifonelli and Roden Biochemical Preparations 12 12 1968]. 

Aprotinin Aprotinin should not be mixed with corticosteroids, heparin, nutrient-containing amino acids or fat emulsions, and tetracyclines. Care must be exercised when diluting solutions of colony-stimulating factors, as they become adsorbed to glass or plastic materials, and the preparation must contain a carrier protein such as albumin to avoid the losses. 

Bone marrow is placed into a balanced salt solution containing preservative-free heparin and a single cell suspension prepared by passing the marrow through a fine wire mesh. Alternatively, a small Potter-Elvehjem tissue homogenizer (Hll) may be used. The nucleated cell count is adjusted to 5-10 X 106 ml of balanced salt solution and deoxyuridine added. Following incubation, 5 p,Ci of tritiated thymidine is added and the mixture incubated for an additional hour. The cells are then washed and the nucleated cell count determined. A 0.1 ml aliquot is then placed on a filter paper disk and dried. The activity of the dried disks is then measured in a scintillation... 

PVPyrH Poly-4-vinylpyridine (PVPyr) prepared by conventional persulfate catalyzed polymerization was dissolved in ethyl alcohol or methyl Cellosolve (2 to 3% solution) for application to polyester film substances. These films adsorb heparin from solutions of sodium heparin to produce anticoagulant surfaces (PVPyrH). Films were painted on, or spread by knife coating. 

Earlier data (3) obtained with tests in cones and chemical determination suggested that the anticoagulant effect of heparin introduced on the wall was greater than that to be expected if all the heparin were dissolved in the plasma. This conclusion requires checking with more uniform heparin layers and improved coagulation test techniques on flat surfaces. It is known that some heparin goes into solution in plasma from GBH or PVPyrH surfaces, as prepared by us, and it seems probable that this is generally true for any adsorbed heparin system in which the... 

Preparation Procedures. Five grains of p-aminopolystyrene (II) prepared as before from cross-linked polystyrene was refluxed with 5% CH3I in CH3OH for 10 hours. The product was filtered, washed with water and methanol, and then dried under vacuum. This quaternary ammonium polymer further reacted with 50 mgm. of heparin in water solution to form the complex (VI). The sulfur microanalysis showed 0.9% by weight. 

The interaction of partially purified, commercial preparations of P. vulgaris with a large number of serum glycoproteins was studied by Morse.105 The lectin precipitated a2-macroglobulin, /3-lipoproteins, and immunoglobulin M. Preparations ofarglycoprotein, orosomucoid, and several immunoglobulin A myeloma proteins also reacted.105 Chon-droitin 4-sulfate, dermatan sulfate, and heparin also precipitated a partially purified preparation of red kidney-bean the precipitation reaction was completely inhibited by 0.5 M sodium chloride solution.3663... 

Meyer and Schwartz, in a study of model compounds structurally related to heparin, showed that the hydrolytic release of sulfate under acid conditions from 2-amino-2-deoxy-D-glucose A -sulfate is more rapid than from D-glucose 6-0-sulfate. These workers also obtained a biologically inactive, nitroso derivative of heparin, and offered this as evidence for a sulfamic acid linkage in the molecule. Wolfrom, Shen and Summers prepared methyl 2-amino-2-deoxy-A-sulfo-tri-0-sulfo-(3-D-glucopyranoside dibarium salt, and found that, on heating a 3 X 10 M solution in 0.004 N hydrochloric acid, the A-sulfate is lost in 20 minutes and the 0-sulfate after 12 hours. 

---