OMB Meeting Book_9-11-14

3

Shoud work in the fume hood and ware personel protective equipment(PPE), For Chloroform should add information that it is“ suspected of causing cancer” to be more awearness during using.

4

Yes, the sample prep for the Phosphatidylinositol Determinations: section C. Hydrolysis, 3) the addition of 50% NaOH. I did not find a way to add the 50% NaOH, that would not cause a violate reaction. Was afraid sample of sample lose or may blow over.

5

No other safety concerns that were not addressed in the written method.

Do you have any information regarding the reproducibility of this method as written? If so, please spec

Question 3

Comment 1

User Comments

No

2

No more data for the reporducibility because for bound inositol the temperature in step heat is just final aglingment to use 130C in May 2014, instead of 110 or 120C.

3 Question 4 Comme t

Need change Base on comment in no. 1 Set a guidance on how to determine the appropriate hydrolysis temperature to ensure complete hydrolysis. Please see the answer to Q2. Based on your experience with the method, are there any recommended changes to the AOAC First Act None.

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3 Question 5

Do you have any general comments about the method?

Comment

User Comments

1

This method required an important instrumentation (2 pumps, a switching valve, 2 CarboPac Columns…). We experiment a gradient and the resolution is as good as with this system. Is it necessary to include in the method the quantification of myo-inositol bound as phosphatidylinositol ? (as we didn’t manage to perform this analysis we don’t know if it is of interest).

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The column-switching technique was a good idea for reducing the run time.

I really like your column switching and I'm looking at possibly modifying our method to use this for other types of products other than infant formula. We analyze all sorts of matrices from clients that contain a lot of things that stick really well to the MA1 column and even require blank injections after to remove anything remaining. The other good thing is this stuff doesn't see the electrode as well when you divert it away Recomment standard to use incase need to prove the %Recovery of boun-inositol analysis need additional add for the SPE step “ All diluent should rinse into centrifuge tube before rinse through the SPE. Need additional guid for Free inositol in step of adjustment to pH 4.5 “To determine the amount of acid that should be added, transfer small amont of sample solution into a small beaker, and insert a pH electrode into the beaker. Add 0.5% hydrochloric acid in 0.5 mL increments until the sample pH is 4.5. Record the amount of acid adde.

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