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Celery Methodology

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Chapter III Methodology
5.1 Study Design
The study will utilize the Quasi experimental research, which describes and analyzes the variables that will be manipulated to influence an effect to the independent variable. Blood will be obtained from students of Medical Technology in Far Eastern University. The blood samples obtained from the student will be observed microscopically after adding the plant extract. Prothrombin time test will also be used to compare the time of coagulation between citrated plasma added with the extract and the citrated plasma alone as reference control.

5.2 Study Setting
5.3 Study Population & Sampling Technique The blood sample will be taken from 30 male or female students of Medical Technology in Far Eastern University. 9 mL of blood will be collected from consented individuals. The subjects will be asked of their medical history regarding if they are not suffering from any blood disorders, diabetes, not a smoker, not taking any medication like heparin, non-steroidal anti inflammatory drugs, and warfarin and free from any bleeding disorders.

Blood Collection First, position the patient’s arm then apply the tourniquet above the antecubital fossa for 3-4 inches. Then, locate the vein and cleanse the venipuncture site with 70% isopropyl alcohol, air-dry. After drying, the tourniquet will be reapplied and the multisample needle will be inserted with the bevel up at an angle less than 30 degrees. Following the correct order of draw, citrated tube will be collected first followed by the non-additive tube and lastly the EDTA tube. 3 ml of blood will be collected from each tube. Remove the tourniquet as soon as blood flow is established. Then, apply direct pressure to the venipuncture site using cotton or gauze pad. The puncture equipment and biohazard wastes will be disposed properly. (Rodaks, 2012)
Collection and Preparation of Apium graveolens extract The celery plant will be bought from a local store in Manila. The plant will be sliced to pieces. The slices will be dried at room temperature for 2 days. After drying, it will be macerated into powder. Two extraction method will be utilized, Soxhlet extraction and decoction. Soxhlet extraction will be followed using methanol as the solvent. The powdered material will be placed in the Soxhlet extractor and will be diluted to different concentrations.

5.4 Data Processing
Microscopic Examination Blood smears will be made using the blood collected from the EDTA tube and non-additive tube. 1 ml of the whole blood diluted with 1 ml of celery extract will serve as the test sample while the 3 ml EDTA with whole blood as the reference control. (Angeles et. al., 2013)

Prothrombin Time The blood collected from the Sodium Citrate tube will be centrifuged at 4000rpm for 15 minutes. 0.5ml of platelet poor plasma (PPP) will be obtained within 30 min to avoid the leakage of cellular components. Three aliquots consisting 50uL of PPP aliquot will be separated, 50uL of the extract with three different concentrations will be mixed and another 100uL of PPP aliquot will be the reference control. The aliquots will be incubated at 37 degrees Celsius for 10 and 3 minutes respectively. 200 ml of the reagent that has been incubated will be added to the aliquots; subsequently the timer will be started. The tubes will be constantly mixed as soon as there is formation of gel, the timer will be stopped and it will be recorded. The procedure will be repeated for three trials (Angeles et. al., 2013)

Microscopic Examination Wedge smears diluted with the extract, stained with Wright’s stain, will be viewed under a microscope to examine the preservation of cell size and shape. It will be compared with the smears made from the EDTA tube as the reference control.
5.5.1 Statistical Analysis The data gathered will be analyed using Analysis of Variance (ANOVA). This test will determine whether the different concentrations of the extract has a significant difference between the variables of the experiment.

5.6 Data Analysis

Prothrombin Time | Sample 1 | Sample 2 | 10% Extract | 13.4 s | 12.9 s | 13 s | 9s | 10s | 12.2s | 30% Extract | 11.1s | 14.6s | 11.1 s | 13s | 11s | 13.2s | 90% Extract | 13.7s | 15.3s | 15.9 s | 13.9 s | 12s | 13.9s | Reference Conrol | 12.1s | 13.4s |

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