Fattah, Miswar
Indonesian Journal of Clinical Pharmacy

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Stabilitas Sampel SOD-Eritrosit dan GPx-Blood dalam Masa Penyimpanan Tujuh Hari Fattah, Miswar; Anggraeni, Sra R.; Alfian, Sofa D.; Levita, Jutti; Diantini, Ajeng
Indonesian Journal of Clinical Pharmacy Vol 1, No 4 (2012)
Publisher : Indonesian Journal of Clinical Pharmacy

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Abstract

Telah dilakukan penelitian mengenai aktivitas SOD eritrosit pada hari ke-0, 1, 3, 5, 7 dengan sentrifugasipada suhu ruangan (22,5 0C) dan pada suhu penyimpanan -80 0C, aktivitas SOD pada hari ke-0 dengan sentrifugasi pada suhu 4 0C, dan aktivitas SOD dengan inkubasi sampel whole blood selama satu hari pada suhu 2–8 0C serta aktivitas GPx hari ke-0, 1, 3, 5, 7 pada suhu penyimpanan 2–8 0C. Penelitian ini menggunakan reagen dari Randox Laboratories yang diukur pada panjang gelombang 505 nm untuk SOD dan 340 nm untuk GPx menggunakan alat Hitachi 917 dari Boehringer Mannheim. Data yang dianalisis menggunakan metode t-test menunjukkan bahwa aktivitas SOD pada hari ke-0, 1, 3, 5, dan 7 dengan sentrifugasi pada suhu ruangan tidak menunjukkan perbedaan yang signifikan, sedangkan pada hari ke-0 dengan sentrifugasi pada suhu 4 0C dan inkubasi sampel whole blood selama 1 hari pada suhu 2–8 0C berbeda secara signifikan. Aktivitas GPx pada hari ke-3 tidak berbeda secara signifikan, sementara aktivitas pada hari ke-0, 1, 5 dan 7 terdapat perbedaan yang signifikan.Kata kunci: Stabilitas enzim, Superoxide Dismutase (SOD), Glutathione Peroxidase (GPx) Stability of Seven Days Sample Storage of Erythrocyte’s SOD and Blood’s GPxAbstractThe research was about SOD erythrocyte activities at day 0, 1, 3, 5, and 7 which centrifuged at room temperature (22.5 0C) and storage temperature (-80 0C), SOD activities at day-0 which centrifuged at 4 0C, SOD whole blood activities with one day incubated at 2-8 0C and GPx activities at day 0, 1, 3, 5, and 7 with 2–8 0C storage temperature. Laboratory analysis were performed by using reagent from Randox Laboratories, and Hitachi 917 analyzer from Boehringer Mannheim. SOD activities were measured at 505 nm absorbance meanwhile 340 nm absorbance is used to measure GPx. Data was analyzed by using t-test method and showed that SOD activities at day 0, 1, 3, 5, and 7 with room temperature centrifuged had no significant differences. Significant differences are found at day-0 with centrifuged at 4 0C and one day incubated whole blood at 2–8 0C. GPx activities at day- 3 had no significant differences. Significant differences are found at day-0,1, 5 and 7 after storage.Key words: Enzyme stability, Superoxide Dismutase (SOD), Glutathione Peroxidase (GPx)
Analisis Kuantitatif 15-F2t-isoprostan dari Plasma Pasien Obstructive Sleep Apnea (OSA) dengan Metode Enzyme Linked Immunosorbent Assay (ELISA) menggunakan Teknik Ekstraksi Imunoafinitas Rusdi, Bertha; Gusdinar, Tutus; Fattah, Miswar
Indonesian Journal of Clinical Pharmacy Vol 1, No 1 (2012)
Publisher : Indonesian Journal of Clinical Pharmacy

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Abstract

15-F2t-isoprostan merupakan penanda stres oksidatif yang kadarnya dalam cairan biologis relatif rendah serta memiliki banyak isomer sehingga diperlukan ekstraksi sampel sebelum dilakukan pengukuran kadar. Metode ekstraksi yang digunakan diantaranya ekstraksi fase solid/Solid Phase Extraction (SPE) serta ekstraksi imunoafinitas. Perbaikan teknik ekstraksi SPE dan teknik ekstraksi imunoafinitas dilakukan untuk membandingkan hasil perolehan kembalinya. Pengukuran dilakukan dengan metode Enzyme Linked Immunosorbent Assay (ELISA). Penilaian efektivitas proses ekstraksi diamati melalui hasil perolehan kembali dari kedua teknik ekstraksi. Teknik ekstraksi dengan perolehan kembali tertinggi digunakan untuk mengukur kadar 15-F2t-isoprostan dari penderita Obstructive Sleep Apnea (OSA). Teknik ekstraksi imunoafinitas menghasilkan perolehan kembali 15-F2t-isoprostan yang cukup baik. Pada penderita OSA kadar 15-F2t-isoprostan dalam plasma cenderung tinggi sehingga memiliki risiko untuk menderita penyakit yang berhubungan dengan aktivitas biologis 15-F2t-isoprostan seperti arteriosklerosis.Kata kunci: 15-F2t-isoprostan, Solid Phase Extraction (SPE), ekstraksi imunoafinitas, ObstructiveSleep Apnea (OSA) Quantitative Analysis of Free 15-F2t-Isoprostane from Plasma of Obstructive Sleep Apnea Patients Using Enzyme Linked Immunosorbent Assay Method with Immunoaffinity Extraction TechniqueAbstract15-F2t-isoprostane is a biomarker in assessment of oxidative stress status that due to its relatively low concentration in biological fluid and also has many isomers, the 15-F2t-isoprostane sample need to be extracted prior to the quantifying processes. Extraction techniques commonly used to extract 15-F2t-isoprostane are solid phase extraction (SPE) and immunoaffinity extraction. Improvements to the SPE and immunoaffinity extraction techniques had been conducted, and the recovery results was then compared. The quantification of 15-F2t-isoprostane then was conducted using Enzyme Linked Immunosorbent Assay (ELISA) method. Then followed by the examination of the plasma recovery results. Extraction technique which had the highest recovery then was used to quantify 15-F2t-isoprostane from plasma of Obstructive Sleep Apnea (OSA) patients. Immunoaffinity extraction technique has a good recovery result. OSA patients have the tendency to have high 15-F2t-isoprostane concentrations in the plasma, therefore have a potential risk to get diseases related to the biological activities of 15-F2t-isoprostane, such as arteriosclerosis.Key words: 15-F2t-isoprostane, Solid Phase Extraction (SPE), immunoaffinity extraction, ObstructiveSleep Apnea