Direct extraction of double-stranded DNA into ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate and its quantification.
Ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate (BmimPF6), as a green solvent, was successfully used for the direct extraction of dsDNA. The extraction efficiency and the distribution coefficient values indicated that trace amounts of DNA at the levels of <5 ng microL-1 facilitate quantitative fast extraction, while proteins and metal species do not interfere. A total of 30% of the DNA in ionic liquid at approximately 20 ng microL-1 was back extracted into aqueous phase in phosphate-citrate buffer with a single-stage extraction. The extraction is demonstrated to be endothermic with an enthalpy of 34.3 kJ moL-1. The extraction mechanisms were proposed and verified by 31P NMR and FT-IR spectra. Interactions between cationic 1-butyl-3-methylimidazolium (Bmim+) and P-O bonds of phosphate groups in the DNA strands take place both in the dissolved BmimPF6 in aqueous phase and at the interface of the two phases. This interaction consequently led to the transformation of DNA conformations, along with a reduction of ethidium resonance light scattering at 510 nm, and a procedure for DNA quantification in ionic liquid was developed based on this observation.
Wang
Jian-Hua
JH
Research Center for Analytical Sciences, Box 332, Northeastern University, Shenyang 110004, China. jianhuajrz@mail.edu.cn
Cheng
De-Hong
DH
Chen
Xu-Wei
XW
Du
Zhuo
Z
Fang
Zhao-Lun
ZL
D002412:Cations / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D007093:Imidazoles / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D012996:Solutions / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared
C412621:1-butyl-3-methylimidazolium hexafluorophosphate; D002412:Cations; D007093:Imidazoles; D012996:Solutions; D004247:DNA