+1 to the better than wallpapering - I thought arguing on the internet was invented to meet all our procrastination needs?
RTMcFadden said:
That difference is electric charge......The goal of the analytical method is that this would be the only difference.
Precisely, that idea is central to all my comments.
RTMcFadden said:
Rule #1 is that you need at least two points to create a curve...... To reliably perform quantitative analysis, I need to operation at or within the limits of this range.
OK. I've been trying to avoid stating my academic background because I've forgotten almost everything I learned....but to save you some time explaining the basics, I do have a science background and I'm not struggling with the maths or concepts of linear systems and experimental variability.......it's the chemistry terminology and techniques that I'm trying to catch up on.
RTMcFadden said:
The concentration portion of your discussion is what a Recovery study would be looking at. This is important to know.
Lets definine
recovery study as meaning figuring out what fraction of EPO from the original sample ends up in the final gel. In which case, yes, absolutely, that whole absorbency versus analyte quantity (y = mx +c) thing would be pretty handy....the more points the merrier within the linear region.....
RTMcFadden said:
Now, where I have a problem is that the data provided indicates that the LLoQ is 125, not 0. This puts the LLoQ inside of the calibration curve that would be generated from the identified control samples, which is an analytical No No. This is why I believe that the positive and negative controls run on the gel are for band identification, not to create the calibration curve...... I can only say that I believe something more is going on that we haven’t been told.
I also think it's primarily about band identification, because the rEPO 'adverse analytical finding' criteria now includes number of bands in the basic area and their relative intensities. It's this kind of point that makes me think the test procedure for rEPO is not a recovery study as per the above definition. It's an analysis of the absorbency ratios of different bands, and a rough estimate of overall concentration by comparing with some other reference analysis. The rough estimate of sample concentration would be used for eliminating results outside the analytical range, and adjusting concentration to facilitate accurate band absorbency analysis. I really find it hard to believe that the standard method for establishing EPO use by an athlete involves tipping some rEPO into the sample.........the lawyers and internet doping apologists would have a field day
. There is also no indication that any analyte is added to the athletes sample, until stability testing stage, that I could find in
http://www.wada-ama.org/rtecontent/document/td2007epo_en.pdf. But as you say, there is more going on than we have been told.....This article doesn't discuss 'adjusting the sample concentration' either....
RTMcFadden said:
f1 = f2 = f3 - these analytes would be proportional to each other, not equal. Yes, this could be a possible scenario, but would require a third marker.
f1,f2 and f3 are not analytes, they are the fraction of EPO from the original sample that makes it onto the gel after centrifuging and double blotting etc. I'm talking about the situation where no rEPO analyte is added. No third marker is required to calculate the ratio of rEPO:nEPO in the gel.
If the goal of the analytical method is met, (f1 = f2 = f3) then rEPO:nEPO in the gel is equal to rEPO:nEPO in the original sample. So stating rEPO in the sample, as a % of tEPO in the sample, is simply another way of expressing rEPO:nEPO in the gel, and does not require the absolute value if tEPO to be known. The accuracy is limited only by the uncertainties in estimating rEPO:nEPO in the gel. Having looked at the example of bands in the above link, I believe uncertainties in rEPO:nEPO could be quite small.....betcha I could write a nice bit of code to do the job.......nice smooth peaks, not too saturated, some little adjustments necessary to account for any overlap......no worries..........