Hi,
I am planning to analyze amino acids using EZ: FAAST kit of phenomenex, by GC-MS and then HPLC. Can you please share your experience with this kit? Is it a good choice or not?
Thanks.
Dear Mahesha,
Amino Acid analysis kits from waters by using UPLC is also good choice.
Hi Dr. Tripathi, thank you for your suggestion. I will go through it.
Mahesha...which kit u used for AA analysis? Kindly share your experience with me.
THanks
Hi I bought EZ:Faast Hydrolysate MS kit. But I used it for FID. I got really excellent results!
i too bought it...i m going to use it for amino acid profile in meat and milk samples...can you please suggest me a sample preparation protocol or a complete protocol. I will highly appreciate it.
Hello everyone,
I used EZfaast (FID) for free amino acid content in meat. But unfortunately, there is no determination of Arg. I need to determine Arg content. Has anyone determined Arg content by EZfaast?
And also has anyone compared EZfaast results to HPLC Amino acid analyser(JEOL etc...) results? Does it have differences?
@ sajid, sorry for delay, i missed your your msg.
I used the same protocol given along with the kit!
I am a trainee in a laboratory which wants to finalize a method of analysis of urinary amino acids. They have the kit EZ : Faast Amino acids Analysis LC/MS. But having followed the procedure to realize the solutions of calibrations and have thrown the analysis, we obtain poor quality results (with peaks which do not look like what is expected). Had any of you also this problem? How can we solve it?
Hello,
By reviewing my message, I realized that I was not very clear. Within the laboratory, they bought the kit for amino acids on the device LC-MS. We realized the analysis of the solution of level calibration 3 by using the data of the kit, with the aim of determining a data bank. The solution of calibration used, is 100 µL of SD1 + 100 µL of SD2 + 100 µL of SD3. We also realized the analysis of the solution of level calibration 1 with volumes of 50 µL. For the rest of the manipulations, we followed the stages explained in the kit.
We realized it, as said previously, to obtain a data bank (the times of retention of every amino acid). Indeed, the laboratory bought the column 2.0 mm while the indications supplied in the kit are for the column 3.0 mm.
The problem it is that the obtained results do not correspond to the profile which we should obtain. Attached, the photo of the graph obtained for the solution of level calibration 3 (that of the solution of level calibration 1 being the same but with peaks of more low intensities).
We tried to relaunch the analysis by changing the parameters or the source (APCI or ESI), but we always obtain a similar graph. To what can be due this problem, in your opinion?
Thank you for the answers that you can bring us. The laboratory assistants and myself are a little lost.
Cheyenne,
They listed the LC conditions for both columns in the kit manual. Also you need run with ESI or ACPI source.
For calibration, the manual is confusing. The way I recommend is dilute the SD1 or SD2 or SD3 to 10,100 times, depending which target amino acid you will test. Then on your sequence, you do the standard curves with conc. and ratio of target/ISTD. for example: you will have 3 sd1 at levels of0.2mM, 0.02mM and 0.002mM. Also you get 3 ratio to ISTD for each amino acid. So you can get a standard curve of ratio vs Conc. with R2~0.998-0.999.
For you chromatography you attached, Is that a Total scan? If yes, you need do a ion extra or run it in EIC or SIM mode.
Hope these help.
Hello, thank you for your help.
The joined photo is the total scan of the mixture of calibration level 3.
Having realized a dilution as you advised it to us, we manage to determine better our peaks. But we had to separate the various SD because together, the graph is illegible. We manage to find all our compounds.
Now our problem is that the times of retention are longer than planned, while we did not change our méhode (we are on the source APCI) and that a peak can contain two sought amino acids.
Thank you again for advice which you bring to us,
Cheyenne
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biochemical Techniques
- Analytical Chemistry
- pH