# HS-SPME Gas Chromatography Approach for Underivatized Acrylamide Determination in Biscuits
## 4 Conclusions  
In this work, we propose a simple methodology for AA quantification based on direct extraction followed by quantification by GC-qMS (HS-SPME/GC-qMS) that does not require a complex sample preparation step.  
The biscuits were grinded and suspended in water for AA extraction from the solid matrix. Addition of propanol to the slurry promoted the AA migration to the headspace facilitating its adsorption on the SPME fiber. GC-qMS in ion extraction mode allowed to achieve a limit of quantification of 91.5 ug/kg of biscuits, which is below the 150 ug/kgbiscuits benchmark level for AA in biscuits for infants and young children. This sequential analysis, comprising extraction and an agent that modifies acrylamide solubility in water, always using the same vial, can be combined with GC-MS, an equipment that is nowadays available in food industries, allowing a simple and quick quality control by producers.  
The following are available online at [[https://www.mdpi.com/article/10.3390/foods10092183/s1](https://www.mdpi.com/article/10.3390/foods10092183/s1)]([https://www.mdpi.com/article/10.3390/foods10092183/s1](https://www.mdpi.com/article/10.3390/foods10092183/s1)), Table S1: List of ingredients used in the biscuit recipe, Table S2: Types of commercially available fiber coatings used in this work, Figure S1: GC-FID chromatogram with acrylamide with a concentration of 5 mg/vial obtained when using a PA fiber. Figure S2. (a) GC-MS chromatograms: (left) standard AA solution (6.4 ug/vial) and (right) biscuits sample with acrylamide detection (9.31-9.30 min); (b) spectrum for AA using the ion extraction chromatography (IEC) mode (71 and 55 \(m/z\) monitoring) and (c) literature full scan spectrum data for propenamide (AA).  
Conceptualization, C.P.P. and M.A.C.; data curation, S.P., A.F.S., A.R., C.N., K.K., Z.C. and S.M.R.; funding acquisition, C.P., Z.C. and M.A.C.; Investigation, C.P.P., A.F.S., A.C.M.N. and D.T.; methodology, C.P.P. and M.A.C.; software, A.R.; supervision, C.N.; validation, C.P.P. and S.P.; writing--original draft, C.P.P.; writing--review and editing, C.P.P., S.P., S.M.R. and M.A.C. All authors have read and agreed to the published version of the manuscript.  
This work received financial support from PT national funds (FCT/MCTES, Portuguese national funding agency for science, research and technology) for the financial support to LAQV  
Figure 6: Acrylamide (AA) content determined in wheat meal biscuits. Different letters represent values that are significantly different (\(p<0.05\)). Dashed lines represent the 350 ug/kg benchmark level for the presence of AA in biscuits and the 150 ug/kg for the benchmark level for the presence of AA in biscuits for infant and young children. * AA biscuit sample also quantified by an external certified laboratory using a conventional HPLC method.  
REQUIMTE (UIDB/50006/2020), CESAM (UIDB/50017/2020; UIDP/50017/2020), and CICECO (UIDB/50011/2020 & UIDP/50011/2020) Research Units through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement, and Compete 2020. This publication was also supported by the Operational program Integrated Infrastructure within the project: Demand-driven research for the sustainable and innovative foods, Drive4 SIFood, 313011V336, cofinanced by the European Regional Development Fund. [PERSON] contract (CEECIND/01873/2017) and [PERSON] post-doc grant (SFRH/BPD/ 117213/2016) was supported by FCT.  
Not applicable.