Use of dried blood or dried matrix spotting for assessment of hematocrit has become a popular laboratory method.
However, the procedure may adversely affect the hematocrit values and thus interfere with the results.
Normal human hematocrit levels are 36 to 48 in adult women and 40 to 52 in adult men.
As the hematocrit rises the blood becomes more viscous and thus the diffusion characteristics of such blood is altered making the spot area variable posing a challenge for assessment.
Blood that is more viscous due to higher hematocrit fails to diffuse through the cellulose paper. However this may be corrected if non-cellulose media is used.
Experiment
Using this non cellulose media various hematocrit levels were assessed.
The hematocrit levels were adjusted as desired by adding or removing plasma from whole blood.
While the original sample had a hematocrit of 45, diluted and concentrated samples contained 20, 30, 45, 65 and 80 serially.
Paroxetine and Nortriptyline were the chosen analytes in the samples.
Results
The samples (15 µl of the 20ng/ml blood sample) were spotted on the Agilent Bond Elut DMS card (p/n A400150).
From each of the dried spots a 3mm disk was punched and the disks were placed onto a 96 well collection plate.
The variation of hematocrit obtained was between 20 and 80. The variability was around 11% in the non cellulose membranes while it was 31% deviated on a cellulose media.
Summary
The study showed that non cellulose media has a significant advantage over cellulose media. The spot variability is smaller with non cellulose media than with cellulose media.
Further cellulose media shows a variability of 50% for hematocrit and paroxetine showed a decrease in the response.
The Agilent Bond Elut DMS card with its non cellulose media outperforms cellulose media by removing the sample variability and improving validation and analysis on the whole.
Further information
About Agilent Technologies
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