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, 2012)
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