Question about calculating label claims on multiple API's

[snalexandria]

Not sure if this qualifies as a "student" level question, if not my apologies.

I am a pharmaceutical biotechnology student currently taking an HPLC course where we utilize Empower 3 software in our lab to run analysis on medication samples. Recently we performed such analysis on a migraine medication with three main peaks or API's (Acetaminophen, Aspirin, and Caffeine). During the processing stage, when filling out the alter sample table, I noticed a limitation wherein we needed to generate an individual result set for each API in order to calculate the respective label claims. This seemed due to the fact that there is only one Sample Weight column in the alter sample table.

Thus, the way we managed this was by plugging in the respective concentration for one API into the Sample Weight column for a specific sample, and its respective standard's concentration in the component editor, then calibrating and quantitating the bracketed standards against the samples to generate such a label claim. A result set was generated for this, then we'd return to the alter sample table and switch the Sample Weight to the next API and reperform the function.

Is it possible to somehow setup the alter sample table in such a way where you can generate a single result set which contains all 3 individual API label claims?

You can see an example of our alter sample table setup here: https://imgur.com/a/kJxPARB

[DR]

Yes. Weight is for sample weight. Label claim for each sample component should also be entered along with your dilution factor.
Click on a Standard in Alter samples, then open up the Component table and enter concentrations for each of your standard components (best way is to add the components with approx. retention times to the processing method, then draw the names into the component table from the processing method - this eliminates the possibility of not calculating something due to differences in spelling).
You should be able to get a mg/mL and %LC result for every component for each sample by processing the run just once.

[snalexandria]

Yes. Weight is for sample weight. Label claim for each sample component should also be entered along with your dilution factor.
Click on a Standard in Alter samples, then open up the Component table and enter concentrations for each of your standard components (best way is to add the components with approx. retention times to the processing method, then draw the names into the component table from the processing method - this eliminates the possibility of not calculating something due to differences in spelling).
You should be able to get a mg/mL and %LC result for every component for each sample by processing the run just once.

Sorry if there was some confusion in how I mentioned it. I am aware on how to calculate a % label claim in regards to adding the sample concentration to sample weight column and filling out the component editor with the standard concentrations. However, its more an issue of differentiating the sample weights.

The issue is the sample in question has 3 sample weights. One for Acetaminophen, one for Aspirin, and one for Caffeine, for each sample as each sample has 3 APIs. What I am trying to accomplish is generating a single result set that calculates an individual % agreement of label claim for all three APIs that don't share identical concentrations.

Currently, the only way we know how to do this is to fill in the sample weight for Acetaminophen, generate a result set, and then return to alter sample, replace the sample weight with Aspirin, generate a second result set, and then repeat of course for Caffeine.

Shouldn't there be a way to enter all 3 sample weight concentrations and generate all three % label claims without unnecessarily generating 3 result sets?

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For some specifics to this case:

Each sample contains these respective theoretical label claim API conc.
Acetaminophen: 1.2 mg/mL
Aspirin: 1.2 mg/mL
Caffeine: 0.312 mg/mL

Our issue being, it would be impossible to apply a 1.2 and 0.312 to the same field in sample weight for concentration. Unless I'm missing something. In any case, appreciate your attempts to help so far!

[DR]

If you're making one injection, you have one sample weight and, in this case, 3 label claim values (one per component), all for the single sample. Once you have a processing method with the 3 components listed in it, the rest is all covered in the components table (a.k.a. Alter Samples). In this table, your standards should have 3 different concentrations listed, and you samples will have one weight and one dilution factor and they should have a label claim for each of the 3 components. One quantitation step for the sample set should yield a result set that has all 3 components listed in it.

[snalexandria]

Hmm. I'll have to wait until I can get my hands back on the software again to take a look at this and perhaps take more photos, but this has not been how our instructor has demonstrated performing this thus far. Though admittedly, they are unsure on how to generate a single result set that contains each % label claim too.

We do add the 3 APIs with respective retention times and such to the processing method during processing/peak integration in order to integrate and label the main peaks for each API. However, when filling out the alter sample window we have always been directed to utilize the sample preparations API concentrations as the source data for inserting the sample weight along with a dilution of 100.

I guess the best question to ask you, is when you refer to a singular "sample weight" per injection where are you sourcing this value from exactly? Our sample weight has always been the API's concentration within the sample in question. This is no issue when there is only a singular API. However, if there are three separate concentrations within the same sample, (where in this case at least 2 are different from one another 1.2 and 0.312) how do you end up with only one singular sample weight in such a case?

[DR]

There are multiple ways to do it.
Look at your dimensional analysis and figure out whether your units all cancel out to deliver the desired result units.

When dealing with finished dosage forms, I input label claim as a weight. I input sample weight, I input a dilution factor and I input a theoretical dosage weight and a standard concentration for each standard component.

From there, it's sample peak area/std peak area*[std]/spl dilution factor*either theoretical or average dose weight. That gets you an assay value in weight.
weight/label claim *100=% label claim.

For multiple APIs in a run, you will have different standard concentrations and label claims for each API and the rest of the math is the same.

It pretty much comes down to opening up alter samples, clicking on your row where you've entered something and using the down arrow to add a new component row. Complete the same info for each new row's new component.
In your method report. right click on white space outside your tables and make sure "summary by all" is chosen.