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Abstrak :
Tujuan penelitian ini adalah untuk mengetahui apakah terdapat perbedaan antara hasil pemeriksaan hematologi rutin dan morfologi darah tepi eritrosit pada sampel darah dengan berbagai konsentrasi antikoagulan Na2EDTA yang berbeda. Penditian ini merupakan penelitian potong lintang. Bahan penelitian berupa 33 sampel darah vena mahasiswa Fakultas Kedokteran UGM Yogyakarta. Dua ml darah dibagi ke datum 4 tabling Na2EDTA yang masing-masing berisi antikoagulan dengan konsentrasi yang berbeda. Tabung pertama berisi Na2EDTA konsentrasi standar, 2 mg/dl, tabung yang lain secara berurutan berisi Na2EDTA dengan konsentrasi 4 mg/dl, 6 mg/dl, and 8 mg/dl. Sebelumnya dibuat sediaan hapus langsung dari setetes darah tanpa antikoagulan (sebagai kontrol) untuk pemeriksaan morfologi darah tepi (MDT). Darah dalam keempat tabung tersebut segera dilakukan pembuatan sediaan hapus dan diperiksa profit hematologi eritrositnya menggunakan SYSMEX SE-9500 automatic analyzer. Terdapat penurunan yang bermakna dari hitung eritrosit, hemoglobin, hematokrit, dan MCHC serta peningkatan yang bermakna dari nilai MCV dan RDW antara konsentrasi Na2EDTA yang berlebihan, sedangkan nilai MCH tidak ada perbedaan. Pemeriksaan MDT menunjukkan perubahan yang bermakna pada bentuk echinocytes serta ditemukan gambaran ghost cells pada sampel darah dengan Na2EDTA yang berlebihan. Disimpulkan bahwa antikoagulan Na2EDTA yang berlebihan akan berpengaruh terhadap morfologi dan beberapa parameter hematologi eritrosit. (Med J Indones 2006; 15:157-64)

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The purpose of this study is to know whether there are differences between hematology profile and morphology of erythrocyles of blood specimens which are prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine Gadjah Mada University. The criteria of subject were male, age from 18 until 22 years old and healthy, ascertained from history taking and vital sign examination. Blood samples from 33 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tube's. Before that, one drop of blood without Na2EDTA anticoagulant was used for making control blood film right after vein puncture. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/dl; the remaining tubes contained consecutively, 4 mg/dl, 6 mg/dl, and 8 mg/dl. Those samples were immediately examined using SYSMEX SE-9500 automatic analyzer for measuring erythrocytes hematological profile and were stained with Wright staining far morphological examination. These procedures were done before 20 minutes of vein puncture. There were significant decrease ofRBC count, HGB, HCT, and MCHC and also significant increase of MCV and RDW between different concentrations of excessive Na2EDTA anticoagulant. MCH did not have significant result. Morphological examination showed significant morphological changes in the form of echinocytes and appearance of ghost cells in the sample treated with excessive Na2EDTA anticoagulant concentration. In conclusion, there are differences in hematological profile and morphology of erythrocytes among blood specimen which are prepared with excessive Na2EDTA anticoagulant in different concentration, except for MCH. Excessive Na2EDTA anticoagulant concentration will affect the blood specimen for peripheral blood examination of erythrocytes by interfering morphology and some of hematological parameters. (Med J Indones 2006; J 5:157-64)

Abstrak :
Akurasi pemeriksaan profil lekosit dipengaruhi beberapa faktor preanalitik diantaranya konsentrasi antiokoagulan. Antikoagulan yang paling sering dipakai pada pemeriksaan darah rutin adalah EDTA. Ketidaksesuaian perbandingan konsentrasi EDTA dengan bahan darah berefek terhadap hasil pemeriksaan darah tepi diantaranya parameter lekosit. Penelitian ini bertujuan untuk mengetahui apakah terdapat perbedaan antara hasil pemeriksaan profil lekosit pada bahan darah dengan berbagai konsentrasi antikoagulan Na2EDTA yang berbeda. Penelitian ini merupakan penelitian potong lintang. Bahan penelitian berupa 33 sampel darah vena mahasiswa Fakultas Kedokteran UGM Yogyakarta. Dua mL darah dibagi ke dalam 4 tabung Na2EDTA. Tabung pertama berisi Na2EDTA konsentrasi standar, 2 mg/dl, tabung yang lain secara berurutan berisi Na2EDTA dengan konsentrasi 4 mg/dl, 6 mg/dl, and 8 mg/dl. Sebelumnya dibuat sediaan hapus langsung dari setetes darah tanpa antikoagulan (sebagai kontrol). Darah dalam keempat tabung tersebut segera dilakukan pembuatan sediaan hapus dan diperiksa profil hematologi lekositnya menggunakan SYSMEX SE-9500 automatic analyzer. Terdapat perbedaan yang bermakna dari hitung lekosit, hitung jenis lekosit absolut dan prosentase monosit pada konsentrasi Na2EDTA yang berlebihan. Prosentase netrofil relatif meningkat dan terdapat perbedaan yang bermakna. Prosentase limfosit, eosinofil dan basofil tidak berbeda secara bermakna. Pemeriksaan morfologi lekosit menunjukkan perubahan yang bermakna berupa tepi sitoplasma yang irreguler, vakuolisasi, dan lobus nukleus yang irreguler akibat pengaruh konsentrasi Na2EDTA yang belebihan. Disimpulkan bahwa penggunaan konsentrasi Na2EDTA yang berlebihan pada preparasi spesimen darah menyebabkan perubahan profil lekosit sesuai peningkatan konsentrasinya. Konsentrasi standar tidak mempengaruhi hitung lekosit dan hitung jenis lekosit serta morfologinya, kecuali berpengaruh terhadap tepi sitoplasma yang irreguler dan lobus nukleus yang irreguler. (Med J Indones 2007; 16:168-75)

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Accuracy of leukocytes profile assessment is influenced by several pre analytical factors, among others, the anticoagulant concentration. EDTA is one of the most frequently used anticoagulant in peripheral blood examination. Several references stated that inappropriate concentration of EDTA anticoagulant in blood sample may affect the result of leukocytes profile in peripheral blood examination. The aim of this study was to evaluate whether there are differences among leukocytes profile in peripheral blood examination specimens, which were prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine, Gadjah Mada University. Blood samples from 30 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tubes. Before that, one drop of blood without Na2EDTA anticoagulant was used to make blood film right after vein puncture, as control. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/ml; the remaining tubes contained 4 mg/ml, 6 mg/ml, and 8 mg/ml respectively. These samples were immediately examined using SYSMEX SE-9500 automatic cell counter to measure the total and differential leukocytes count; and were stained with Wright staining for morphological examination under the microscope. These procedures were done before 20 minutes of vein puncture. There were significant decrement of total leukocytes count, absolute differential leukocytes count and monocyte percentage following excessive Na2EDTA administration. Neutrophil percentage was found to be relatively increased and the difference was significant. Lymphocyte, eosinophil and basophil percentages were not significantly different. Morphological examination showed significant increment in irregular cytoplasm margin, vacoulation and irregular nuclei lobes following excessive Na2EDTA administration. It is concluded that excessive concentration of Na2EDTA used in blood specimen preparation, will lead to changes in leukocytes profile as the concentration increased. Standard Na2EDTA anticoagulant concentration did not alter any leukocytes count and morphology, except for irregular cytoplasm margin and irregular nuclei lobes. (Med J Indones 2007; 16:168-75)