Apparatus for molecular weight separation
- Richland, WA
- Haverhill, MA
The present invention relates generally to an apparatus and method for separating high molecular weight molecules from low molecular weight molecules. More specifically, the invention relates to the use of microdialysis for removal of the salt (low molecular weight molecules) from a nucleotide sample (high molecular weight molecules) for ESI-MS analysis. The dialysis or separation performance of the present invention is improved by (1) increasing dialysis temperature thereby increasing desalting efficiency and improving spectrum quality; (2) adding piperidine and imidazole to the dialysis buffer solution and reducing charge states and further increasing detection sensitivity for DNA; (3) using low concentrations (0-2.5 mM NH4OAc) of dialysis buffer and shifting the DNA negative ions to higher charge states, producing a nearly 10-fold increase in detection sensitivity and a slightly decreased desalting efficiency, (4) conducting a two-stage separation or (5) any combination of (1), (2), (3) and (4).
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- DOE Contract Number:
- AC06-76RL01830
- Assignee:
- Battelle Memorial Institute (Richland, WA)
- Patent Number(s):
- US 6187190
- OSTI ID:
- 873547
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
molecular
weight
separation
relates
method
separating
molecules
specifically
microdialysis
removal
salt
nucleotide
sample
esi-ms
analysis
dialysis
performance
improved
increasing
temperature
desalting
efficiency
improving
spectrum
quality
adding
piperidine
imidazole
buffer
solution
reducing
charge
detection
sensitivity
dna
concentrations
0-2
nh4oac
shifting
negative
producing
nearly
10-fold
increase
slightly
decreased
conducting
two-stage
combination
detection sensitivity
molecular weight
buffer solution
weight molecules
separation performance
reducing charge
weight separation
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