The comparison of five precipitation methods for the isolation of HDL
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Tarih
2001
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Dergi ISSN
Cilt Başlığı
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Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Yüksek Dansiteli Lipoprotein (HDL) separasyonunda kullanılan dört çöktürme yöntemi [Dekstran sülfat 500,000 (DS)-MgClsub2/sub, polietilen glikol (PEG) 6000, PEG 20000, fosfotungstat (PTA)-MgClsub2/sub] heparin-MnClsub2/sub yöntemi ile karşılaştırıldı, Lipoprotein çökmesi üzerine reaktif konsantrasyonunun, inkübasyon ve santrifügasyon süresinin etkileri incelendi. 21 serum örneğinde; son MnClsub2/sub konsantrasyonun 91 mM olarak kullanılmasıyla heparin-MnClsub2/sub yöntemiyle 35.57 mg/dL ortalama HDL konsantrasyonları elde edildi. Son MgClsub2/sub konsantrasyonu 45 mM olan DS ile (son konsantrasyonu 0.909 g/L) heparin-MnClsub2/sub karşılaştırıldığında biraz düşük (-1.02 mg/dL)değerler bulundu. Oysa PEG 6000 (1.43 mg/dL) ve PTA-MgClsub2/sub (1.82 mg/dL) ile elde edilen sonuçlar hafifçe yüksekti. Aynı şekilde PEG 20000 ile elde edilen sonuçlar heparin-MnClsub2/sub ile elde edilenlerden yüksekti (0.98 mg/dL). PTA-MgClsub2/sub ile çöktürme yönteminin, diğer yöntemlerle mukayese edildiğinde, reaktif konsantrasyonu ve ısı değişikliklerine daha duyarlı olduğu gözlendi. Sonuçta tüm bu çöktürme yöntemlerinin hepsinin yeterli özgüllüğe, tekrarlanabilir/iğe ve duyarlığa sahip oldukları, hem klinik analizlerde hem de büyük çaplı epidemiyolojik çalışmalarda rahatlıkla kullanılabilecekleri tespit edildi.
We compared four precipitation methods [dextrane sulfate 500,000 (DS)-MgCl<sub>2</sub>, polyethylene glycol (PEG) 6,000, PEG 20,000, phosphotungstate (PTA)-MgCl<sub>2</sub>] for separation of serum high-density lipoprotein (HDL) with heparin-MnCl<sub>2</sub> methods. The effect of reagent concentration, incubation and centrifugation time on lipoprotein precipitation was assessed. In 21 serum samples, heparin-MnCl<sub>2</sub> method with final MnCl<sub>2</sub> at 91 mM produced results with mean HDL concentration of 35.57 mg/dL. Again results with DS (final concentration 0.909 g/L) at final MgCl<sub>2</sub> concentration of 45 mM averaged slightly lower (-1.02 mg/dL) than the comparison heparin-MnCl<sub>2</sub> method. While results that were obtained by the PEG 6,000 (1.43 mg/dL) and PTA-MgCl<sub>2</sub> (1.82 mg/dL) were slightly higher. Results by a PEG 20,000 precipitation method were again higher than heparin-MnCl<sub>2</sub> (0.98 mg/dL). Precipitaiton by PTA-MgCl<sub>2</sub> appeared more sensitive to reagent concentration and temperature variations than the others. We conclude that these all precipitation methods have enough specificity, precision and sensitivity and can be used in clinical analyses as well as in large scale epidemiological studies with satisfactory results.
We compared four precipitation methods [dextrane sulfate 500,000 (DS)-MgCl<sub>2</sub>, polyethylene glycol (PEG) 6,000, PEG 20,000, phosphotungstate (PTA)-MgCl<sub>2</sub>] for separation of serum high-density lipoprotein (HDL) with heparin-MnCl<sub>2</sub> methods. The effect of reagent concentration, incubation and centrifugation time on lipoprotein precipitation was assessed. In 21 serum samples, heparin-MnCl<sub>2</sub> method with final MnCl<sub>2</sub> at 91 mM produced results with mean HDL concentration of 35.57 mg/dL. Again results with DS (final concentration 0.909 g/L) at final MgCl<sub>2</sub> concentration of 45 mM averaged slightly lower (-1.02 mg/dL) than the comparison heparin-MnCl<sub>2</sub> method. While results that were obtained by the PEG 6,000 (1.43 mg/dL) and PTA-MgCl<sub>2</sub> (1.82 mg/dL) were slightly higher. Results by a PEG 20,000 precipitation method were again higher than heparin-MnCl<sub>2</sub> (0.98 mg/dL). Precipitaiton by PTA-MgCl<sub>2</sub> appeared more sensitive to reagent concentration and temperature variations than the others. We conclude that these all precipitation methods have enough specificity, precision and sensitivity and can be used in clinical analyses as well as in large scale epidemiological studies with satisfactory results.
Açıklama
Anahtar Kelimeler
Genel ve Dahili Tıp
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Gülhane Tıp Dergisi
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Cilt
43
Sayı
3
Künye
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