December 28, 2024, 10:58:40 AM
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Topic: Over quantification of Cholesterol in Cheese with AOAC method(45.4.10_994.10)  (Read 4484 times)

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Offline SiberiaFrogUnit

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Hi, I work in Cheese making factory and I'm a person in charge of nutrition compound analyzing of cheese.

Our country has no forum like here, so I'm writing my question at here.

2 months ago, I found that some product's cholesterol amount is over than spec.
So I sent samples to our company's central laboratory.

But central laboratory's results were just a half of mine. What's the problem?

We both used same method(AOAC 45.4.10, 994.10 Cholesterole in Foods).
I did almost same as AOAC method.

Differences were,
1. use 99.99% EtOH HPLC grade(in method, use 95% EtOH)
2. didn't use silanize tube(when I used this method first time, I used both silanize tube and normal tube. But their results had no significant difference)

what's wrong with me??

Anyone who has any opinion, I don't care that is right or not, Just tell me about your idea freely.
If need more information, reply to me. I will answer ASAP.

Offline Arkcon

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We do have someone on this board who also performs analysis on cheese in the EU, so you are in luck.  They may be available shortly for more specific help.  However, whenever you make any change to a method, you can't just say it makes no difference once, you must validate the new method.  Basically, you must formulate an experimental plan, try many known samples -- some standards that are traceable back to another method, possibly even controlled by a regulatory body, and also some known and spiked samples -- perhaps 3 or more separate production lots, and have the statistics on the data to support that the new method gives identical results.  This should all be documented, and available for a regulatory body to review at their request.

The sudden result giving double values is very suspicious for me.  Perhaps you have made other changes, that even you are not aware of.  Small, minor changes in a method should only produce slight changes. A quick review of this method shows many wet chemical steps before the GC analysis, so it might be hard to pin down where the method might go wrong.  Logically, failure to silanize tubes should produce less cholesterol signal, because it does tend to bind to plastic tubes.  But analytical chemists often don't have the luxury of making determinations on what the "figure" ought to arise logically.  Instead, we have to rely on what we can prove empirically.
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline marquis

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Just as a sanity check, please double check your math. 

As Arkcon said, the factor of two is suspicious.  On possible cause could be a calculation error.

Offline SiberiaFrogUnit

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thanks, Arkcon, marquis.
I'll re-check all of procedures and calculations.
I think I need to open all of my results and calculating to find my fault more easily.
Now I'm working, so I will update all of mine after back to home.
Thanks you all.

Offline curiouscat

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I'll second the calculation check.

Also, can you re-run  another old sample that you've previously analysed?

Offline SiberiaFrogUnit

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I'm sorry for late. We had one serious problem. We all had to focus to that. And this problem is still ~ing... :'(

Here is my results and raw data.

And I re-checked all of my calculation and I'm sure that I used this formula.
the formula what I used is,

Cholesterole contents(mg/100g) = (ppm*/1000) x (3x100) / (sample weight(g)x( 25 / 100 ))

* Raw data's ppm result. Obtained from correlation curve.

Standard solution and ISTD solution were made everytime I perform analysis.

thanks you curiouscat.
I can re-run another old sample that I analyzed. But I'm sorry that I have no time to do that untill next week.

again, sorry for late.

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