Comparison effects of mannitol and hypertonic saline on coagulation in vitro: a randomized, blinded trial.

We will take 20 ml blood ( 5 tube of 4ml) an antecubital vein from healthy volunteers for study.. Four ml blood sample was seperated for each group. In each group 2 ml blood were collected into polypropylene tubes (Vacuette, Greiner Bio-one, Austria) containing 3.2% buffered citrate, giving a volume ratio of 1:10 for ROTEM analysis and 2 ml blood were collected into EDTA tubes containing 1.2 mg anhydrous EDTA per 1ml blood (Vacuette, Greiner Bio-one, Austria) for hemogram analysis.Immediately after sampling all blood samples were diluted with the test solutions.Thereafter, within 2 hours after blood withdrawal, thromboelastometry coagulation analysis (ROTEM; Pentapharm Co., Munich, Germany) was made in random order for all samples. The coagulation process will initiate by recalcification (Star-TEM; Pentapharm Co., Munich,Germany) and activation with tissue factor EXTEM and INTEM for monitoring the extrinsic system, and intrinsec system . In the FIBTEM assay (fib-TEM, Pentapharm Co.) a plateletinhibitor (GPIIb/IIIa platelet receptor antagonist abciximab) together with cytochalasin D was added to assess the contribution of fibrinogen to clot strength, and EXTEM analysis was carried out. Automatically measured thromboelastometry parameters of blood coagulation will be recorded. This parameters are Clotting time (CT), Clot formation time (CFT) and Maximum clot firmness (MCF). (We will record each CT,CFT and MCF measurements at INTEM, EXTEM and FIBTEM) We also will record hemoglobin and platelet value of each sample. In present study there are five arm. Arm 1: Control group. Sample contain 2 ml pure blood. We will make thromboelastometry coagulation analysis this sample. Arm 2 : 7% Mannitol group. Sample contain: 1.86ml blood and 0.14 ml 20% mannitol. We will make thromboelastometry coagulation analysis this sample. Arm 3: 7% hypertonic saline group. Sample contain 1.86 ml blood and 0.14 ml 3%NACL. We will make thromboelastometry coagulation analysis this sample. Arm 4: 6% Mannitol - 8%HES group. Sample contain 1.72 ml blood, 0.12 ml 20% mannitol and 0.16 ml Hydroxyethyl Starch 130/0.4. We will make thromboelastometry coagulation analysis this sample. Arm 5: 6% Hypertonic saline - 8%HES group. contain 1.72 ml blood, 0.12 ml 3%NACL and 0.16 ml Hydroxyethyl Starch 130/0.4. We will make thromboelastometry coagulation analysis this sample.