Analysis of Acrilamide by HPLC C18

Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.

8 posts Page 1 of 1
Dear all,
I am trying to set up a method for acrylamide in potato chips analysis but I don't known what is going wrong because I don't have much success. My idea is to detect acrilamide in potato chip at 100 ng/g and conduct the quantification with matrix-matched calibration curve. I have tried diferent HPLC methods with C18 columns, however, my feeling is that the acrilamide is not retained by the column and the different Rts indicated depend on the length of the peak tube. I finally manage to obtain a more o less gaussin peak using a zorbax C8 column.

On the other hand I tried to extract acrilamide with water, hot water, quechers, and SPE but the reproducibility is not very good, my RSD is about 60% for 500 ng/g.

Does anybody known a method that it is realiable and reproducible at 100 ng/g level?

Thank you very much in advance,
Carol
Hi Carol,

Feel free to review this article:

https://www.sciencedirect.com/science/a ... 7304019715

Journal of Chromatography A, Volume 1088, Issues 1–2, 23 September 2005, Pages 193-199

Abstract
A simple and rapid method using liquid chromatography coupled to diode array detection (LC–DAD) was developed for the determination of acrylamide in potato-based foods at low levels. The method entails extraction of acrylamide with methanol, purification with Carrez I and II solutions, evaporation and solvent change to water, and cleanup with a Oasis HLB solid-phase extraction (SPE) cartridge. The final extract was analyzed by LC–DAD for quantification and by liquid chromatography coupled to mass spectrometry (LC–MS) for confirmation. The chromatographic separations were performed on a hydrophilic and a hydrophobic interaction columns having good retention of acrylamide under 100% aqueous flow conditions (k′ 3.67 and 2.54, respectively). The limit of quantitation was estimated to be 4.0 μg/kg based on the signal-to-noise ratio of 3 recorded at 226 nm. Recoveries of acrylamide from potato chips samples spiked at levels of 250, 500 and 1000 (n = 4 for each level) μg/kg ranged between 92.8 and 96.2% with relative standard deviations of less than 5%. The results of this study revealed that a conventional LC instrument coupled to DAD can also be used accurately and precisely, as an alternative to tandem LC–MS methods for the determination of acrylamide in potato-based foods.
MattM
This one may also be of interest:

https://www.sciencedirect.com/science/a ... 7314011996

A new derivatization approach with d-cysteine for the sensitive and simple analysis of acrylamide in foods by liquid chromatography–tandem mass spectrometry

Abstract
A liquid chromatography–tandem mass spectrometry method (LC–MS/MS) was developed in order to determine the amount of acrylamide in foods after derivatization with d-cysteine. The sulfhydryl group of d-cysteine was added at the β-site double bond of acrylamide to form 2-amino-3-(3-amino-3-oxo-propyl)sulfanyl-propanoic acid. Deuterated acrylamide (acrylamide-d3) was chosen as the internal standard (IS) for analyzing the food samples. Acrylamide was extracted from 2.0 g of food sample with 6 mL of methylene chloride, and the organic extract was diluted with 3 mL of hexane, and then the analyte was back-extracted with 0.5 mL of pure water. The derivatization of acrylamide was performed in the water extract. The best reaction conditions (3.0 mg of d-cysteine, a pH 6.5, a reaction temperature of 90 °C, and a heating time of 50 min) were established by the variation of parameters. The formed derivative was injected into the LC–MS/MS without further extraction or purification procedures. Separation and detection were improved with the use of an ion-pairing reagent of perfluorooctanoic acid. Under the established conditions, the limits of detection and the limits of quantification were 0.04 μg/kg and 0.14 μg/kg, respectively, and the inter-day relative standard deviation was less than 8% at concentrations of 20 and 100 μg/kg. The method was successfully applied to determine the amount of acrylamide in potato chips, French fries, and coffee.
MattM
Hi,

Thanks a lot for your recomendation,

Did you try any of the method? There are several methods over the internet, however I would like one that somebody has used and verified that it works,

Thanks,
Carolina
Hi Carol,

No, these merely are abstracts of J. Chrom. A articles which seem to describe procedures that meet the criteria you are interested in achieving. I have not verified/validated either of these methods through an experiment on my own.

Perhaps the authors of these papers could be contacted and they could share with you further relevant details of the work they have published? I have had success with doing this in my past.

Best wishes in obtaining the information that you seek!
MattM
@ Rndirk,

Thank you or providing for the pointer above, there seem to be some relevant ideas within.
MattM
Some years ago we were contacted by someone from a company that was manufacturing polyacrylamides for use in contact lenses. The government was requiring them to report the content of unreacted acrylamide monomer. The customer had read that our company's size exclusion chromatography (SEC) columns can separate analytes the size of amino acids, and he wanted to know if they could also separate acrylamide monomer from the dimer and higher oligomers. It turned out that they could. The mobile phase was 50 mM formic acid. Maybe this would be a convenient way for you to perform this analysis.
PolyLC Inc.
(410) 992-5400
aalpert@polylc.com
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