Proteinleri Ayırmak İçin Ters Faz Kromatografisi Kolon Dolgu Materyali Tasarımı: Bölüm 2- Kromatografik Performans

İki bölümden oluşan çalışmanın ikinci bölümünde çalışmanın ilk bölümünde sentezlenmiş olan eş boyutlu ve gözenekli poli(vinil asetat-co-divinilbenzen) [poli(VAc-co-DVB)] ve poli(divinilbenzen) [poli(DVB)] kolon dolgu materyalleri Ters Faz Kromatografisi (Reversed Phase Chromatography, RPC) için farklı koşullarda test edilmiştir. Sentez sonrası elde edilen partiküllerle doldurulan kolonların kromatografik performansları incelediğinde, poli(VAc-co-DVB) (2500 tabaka/m) ve poli(DVB) (1500 tabaka/m) kolon için literatürdeki değerlerden daha yüksek ve kararlı teorik tabaka sayısı değerleri elde edilirken pik çözünürlük değerlerinin de kabul edilen değerlerin üstünde olduğu görülmüştür. Poli(VAc-co-DVB) (%93-99) ulaşılan protein geri kazanım değerleri ise bütün proteinler için kantitatife yakın değerler olarak belirlenmiştir. Üretilen kolonlar için analitlerin, analizden analize ve günden güne tekrarlanabilirlik testleri yapılmış, hem “analizden analize “ hem de “günden güne” Bağıl Standart Sapma (BSS) değerleri % 2,8’in altında bulunmuştur.

Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance

In the second part of the two-part study, monodisperse and porous poly(vinyl acetate-co-divinylbenzene) [poly(VAc-co-DVB)] and poly(divinylbenzene) [poly(DVB)] column packing materials synthesized in the first part of the study were tested in Reverse Phase Chromatography (RPC) under different conditions. The investigation of column performances concluded that columns packed with poly(VAc-co-DVB) (2500 plate/m) and poly(DVB) (1500 plate/m), showed high and stable theoretical plate number and high peak resolution values compared to literature values. Protein recovery data in RPC column, poly(VAc-co-DVB) (93-99%), were determined as similar as quantitative values for all proteins. Reproducibility tests of analytes of poly(VAc-co-DVB) column were assessed "run to run" and "day to day" Relative Standard Deviation (RSD) values were determined under 2.8%.

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