Extraction And Determination Of Acrylamide In Polyacrylamide Gel By High Performance Liquid Chromatography

Objective To establish a method of extraction and determination of acrylamide (AM) in polyacrylamide gel (PAMG) by High Performance Liquid Chromatography (HPLC), which should be simple, accurate, and suitable for the determination of acrylamide in polyacrylamide gel. And then detect the samples of polyacrylamide gel and that of drawing from bodies.Methods To compare the recoveries of three pretreatment methods for polyacrylamide gel samples include microwave assisted extraction, water extraction, and accelerated solvent extraction. Pretreating the polyacrylamide gel by the method of water extraction, filtrating the extracts with 0.45μm PVDF film, cleaning it up with Oasis HLB solid-phase extraction (SPE) cartridge and Bond Elut-Accucat SPE cartridge, the final extract was quantified by HPLC.Results The recoveries of pretreatment methods of microwave assisted extraction, water extraction, and accelerated solvent extraction are 44.2~70.0%, 87.8~94.9%, and 36.2~64.2%. The chromatographic conditions include Angilent TC-C18 column, 100% water with the flow rate of 1.0mL/min used as mobile phase. Linear relationship between peak area and concentration of acrylamide was obtained in the range of 0.05~100μg/mL (R2=0.9997). The limit of detection (LOD) and the limit of quantitation (LOQ) were 0.05μg/mL and 0.15μg/mL in the basis of signal to noise ratios of 3:1 and 10:1, respectively. Relative standard deviations were 4.1% and 5.3% in within-day (n = 6) and day-to-day (n = 8) precision tests, respectively. The concentrations of acrylamide in Polyacrylamide Gel are in range of 0.2~0.8μg/ml, and that of drawing from bodies are all lower than detection limits.Conclusion The pretreatment method of water extraction is a simple, effective pretreatment method. So the method which the sample was soaked in quantitative water was chosen in this study. On the basis of characteristics of polyacrylamide gel and solubility of acrylamide in water, polyacrylamide gel could achieve swelling state completely in superfluous water. Then the concentration of acrylamide formed equilibrium between the water inside and outside the hydrogel. Subsequently, the precise content of acrylamide in polyacrylamide gel was calculated by means of determination of acrylamide in the extractant. Not only were sample preparation procedures simplified greatly but also a satisfactory result of recovery was obtained through replacement of detecting matrix. This HPLC method is simple, accurate and suitable for the determination of acrylamide in polyacrylamide gel. The detected data of polyacrylamide gel indicate that polyacrylamide gel is stable relatively and do not degrade to a great deal of acrylamide monomer in complicated environment in vivo. And the content of acrylamide in polyacrylamide gel removed from human bodies is lower than that of the polyacrylamide gel product. It is possible that the water in the gel can move freely in the complicated environment in vivo. So the acrylamide can go with water freely. And then acrylamide will be degraded by bodies. Consequently,toxicity of acrylamide monomer may not be the major factor causing postoperative complications. Maybe the property of polyacrylamide gel causing allergy is relative with postoperative complications. As a result, Further studies of the gel's microstructure and long-term change are still needed. And the better methods of determination of acrylamide in polyacrylamide gel are needed to be developed, for the content of acrylamide detected by this method is lower than the limit of detection.