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Analysis of acrylamide in food: dedicated to Professor Dr. Werner Baltes.

Since the first discovery of the presence of acrylamide in a variety of food products in April 2002, numerous methods have been developed to determine the acrylamide monomer in heat-treated carbohydrate-rich food. These detection methods are mainly MS-based, coupled with a chromatographic step using LC or GC. The Food Chemistry Institute (LCI) of the Association of the German Confectionery Industry (BDSI) therefore established a detection method by means of aqueous extraction plus a cleaning step and LC-MS/MS detection, making great efforts to ensure internal and external validation. Citing potato crisps as an example, we will in the following show how the German manufacturing companies have gone to great pains to reduce acrylamide levels in their products.

Acrylamide↗

2',3'-Cyclic nucleotide 3'-phosphohydrolase in acrylamide-induced optic neuropathy in rabbit.

The in vivo effects of acrylamide on the nerve tissues of the rabbit optic pathway were examined. The optic nerve of acrylamide-treated rabbits showed inflammatory changes and degeneration. The decrease of myelin was observed with myelin stain. The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC.3.1.4.37) which is firmly bound to myelin was markedly diminished by acrylamide intoxication.

2',3'-Cyclic Nucleotide 3'-Phosphodiesterase↗

The neurotoxicity of acrylamide: an overview.

This paper summarizes recent research conducted on the neurotoxicity of acrylamide. Of particular interest is research concerning the possible effects that acrylamide might have on the central nervous system. Neurochemical and psychopharmacological studies indicating that acrylamide affects neurotransmitter systems involved in the mediation of behavior are reviewed. Needs for future research are discussed.

Acrylamides↗

Conditioned taste aversion following acutely administered acrylamide.

Fischer-344 rats were acclimated to a daily 15 min period of water availability. After water consumption had stabilized, the rats were permitted a 15 min access to 0.15% (w/v) solution of saccharin. One to two min following saccharin presentation, they were given various doses of acrylamide (10-50 mg/kg) or lithium chloride (LiCl; 3 mEq/kg) by gavage. Three days later, aversion to the saccharin solution was determined. Acrylamide, like LiCl, produced an aversion to saccharin. The effect of acrylamide appeared to be dose-related with the threshold dose being 10 mg/kg.

Acrylamides↗

Interactions between "handling" and acrylamide on endocrine responses in rats.

Serum prolactin and corticosterone levels were measured in rats that had been gentled by handling and that had been exposed to 100 mg/kg acrylamide. Handling reduced both corticosterone and prolactin levels. Acrylamide had no effect on corticosterone levels. Prolactin levels were significantly lower in acrylamide-exposed nonhandled animals. These findings are discussed in terms of their implications for studying the effects of toxicants on neural function.

Acrylamides↗

Comparison of accelerod and rotarod sensitivity in detecting ethanol- and acrylamide-induced performance decrement in rats: review of experimental considerations of rotating rod systems.

The relative sensitivity of two rotating rod techniques in detecting performance decrement in rats was assessed after treatment with either ethanol or acrylamide. Performance on the rod during acceleration at approximately 1 rpm/sec (accelerod) was compared to that obtained on the same rod operated at a constant speed of 20 rpm (rotarod). Rats trained to either task received a single oral dose of ethanol (0.5, 1.0, 1.5, or 2.0 g/kg) or a series of ip doses of acrylamide (25 or 50 mg/kg/day) before testing. Accelerod performance was significantly more disrupted at lower doses and for longer periods of time after ethanol ingestion than was rotarod performance. Likewise, task disruption resulted from repeated injections of acrylamide also appeared at lower cumulative doses using the accelerod. A higher proportion of the native subjects were successfully trained and the mean time for training to minimum performance standards was significantly less using the accelerod. The greater sensitivity of the accelerod technique in detecting neurotoxic effects is attributed primarily to the fact that this test provides a continuous measure of the upper limit of performance rather than the quantal or arbitrarily truncated measure that one usually obtains from the rotarod. In light of these findings, a review is presented of the primary experimental variables that affect the reliability of data obtained in neurotoxic evaluations using the accelerod and rotarod procedures.

Acrylamides↗

Subchronic toxicity of acrylamide administered to rats in the drinking water followed by up to 144 days of recovery.

Groups of male and female Fischer 344 rats were administered acrylamide in their drinking water at 0, 0.05, 0.2, 1, 5, or 20 mg/kg/day for up to 93 days. Following the administration of acrylamide in the drinking water, male rats from each dose level were held for up to 144 days of recovery. The 20 mg/kg/day groups had definite treatment-related effects after 92 (males) and 93 (females) days. They were dragging the rear limbs, body weights were decreased, serum cholinesterase activity was decreased in top dose females, and packed cell volume, red blood cell, and hemoglobin values were slightly decreased in males and females. In the 20 mg/kg/day groups, the primary target tissue was the peripheral nerve with lesions consisting of severe degeneration characterized by demyelinization and axonal loss. Slight spinal cord degeneration was observed. Other effects included atrophy of skeletal muscle, testicular atrophy, and distended urinary bladders; these were probably secondary to the nerve degeneration. After 144 days of recovery, the lesions had partially or completely reversed. Parameters affected at the 5 mg/kg/day dose level after 92 (males) and 93 (females) days consisted of peripheral nerve degeneration which were of a lesser degree of severity than those seen in the 20 mg/kg/day groups and appeared to have completely reversed after 111 days of recovery. In rats given 1 mg/kg/day, a minimal treatment-related effect was observed in males after 92 days, and this was limited to very slight nerve degeneration using electron microscopy (females were not examined by electron microscopy). This observed effect appeared to have reversed after 25 days of recovery. No treatment-related effects were seen in any of the parameters monitored in the rats given 0.05 or 0.2 mg/kg/day of acrylamide.

Acrylamides↗

Confocal microscopy of thick sections from acrylamide gel embedded embryos.

The preparation of optically clear, thick sections of fragile embryonic tissues greatly aids the power of confocal scanning laser microscopy in imaging three-dimensional structures. We report here conditions for embedding, sectioning, and staining embryos in polyacrylamide gels for a variety of confocal imaging techniques. Infiltration of tissues in standard mixtures of 10-15% acrylamide monomer yields, upon polymerization, blocks that cut easily by vibratome between 50 and 1,000 microns. These conditions worked well for tissues previously stained or for staining gel sections with low molecular weight water-soluble fluorochromes (MW < 5 kD [e.g., propidium iodide, phalloidin]). For immunostaining of tissue after embedding and sectioning, the acrylamide concentration was reduced to 2-3% acrylamide to allow access of immunoglobulins to antigenic sites; such gels were supplemented with 1% agarose to facilitate sectioning and handling. Either method yielded abundant, optically clear, and easily handled sections for mounting and examination in water-miscible media.

Acrylic Resins↗

Water-soluble poly(acrylamide-allylamine) derivatives of saccharides for protein-saccharide binding studies.

Water-soluble poly(acrylamide-allylamine) copolymers containing covalently bound amino groups, prepared by copolymerization of acrylamide and allylamine, can be used as general carriers for coupling of different types of saccharides or saccharide derivatives. The water-soluble macromolecular carbohydrate derivatives can be easily labelled and used in various solid-phase techniques to study protein-saccharide interaction. Two types of coupling reaction were used to prepare polyacrylamide derivatives of saccharides: reductive amination was applied to couple the reducing disaccharides and a carbodiimide reaction was used to couple heparin via its carboxyl groups to the amino groups of the poly(acrylamide-allylamine) derivative. Peroxidase labelled or biotinylated derivatives were shown to be useful in studies on the binding properties of lectins and proteins from boar seminal plasma.

Acrylic Resins↗

Adsorption of bovine serum albumin onto acrylamide-maleic acid hydrogels.

In this study, acrylamide-maleic acid hydrogels containing different amounts of maleic acid have been prepared by irradiating with gamma radiation. They have been used in experiments on bovine serum albumin (BSA) adsorption. Acrylamide-maleic acid hydrogel containing 60 mg of maleic acid and irradiated at 5.20 kGy has been used for swelling and diffusion studies in water, solutions of NaCl and BSA. For this hydrogel, maximum and minimum swellings have been observed with water (1680%) and the solution of BSA (1085%), respectively. Diffusions of water, NaCl and BSA within hydrogels have been found as non-Fickian types of diffusion. In the experiments of BSA adsorption, Langmuir type adsorption has been found. The hydrogel prepared with 60 mg maleic acid and having been irradiated at 2.00 kGy has been found to be the best adsorption system for BSA. The adsorption capacity of acrylamide hydrogel has been increased by the addition of maleic acid almost 1.5-3-fold.

Acrylic Resins↗

Electrophoretic characterization of the nonspecific esterases of the mosquito, Culex tarsalis: conventional and isoelectric focused acrylamide gels.

1. The nonspecific esterases of the mosquito, Culex tarsalis, were examined through conventional and isoelectric focusing acrylamide gel electrophoresis. 2. Conventional acrylamide gel electrophoresis resolved five components. These were characterized as: three carboxylesterases, one acetylcholinesterase and one acetylesterase. 3. Isoelectric focusing resolved 18 components. These were characterized as: 14 carboxylesterases, two acetylcholinesterases, one acetylesterase and one arylesterase. 4. The reproducibility and reliability of isoelectric focusing is discussed and compared to conventional acrylamide gel electrophoresis for the examination of multi-component isozyme systems such as non-specific esterases.

Animals↗

Dominant lethal mutations, heritable translocations, and unscheduled DNA synthesis induced in male mouse germ cells by glycidamide, a metabolite of acrylamide.

The hypothesis that acrylamide induces dominant lethal mutations and heritable translocations in male mice, not through direct adduction, but by conversion to the reactive epoxide, glycidamide, was investigated. Three studies, namely, induction of dominant lethal mutations, heritable translocations, and unscheduled DNA synthesis in spermatids, which were conducted earlier in this laboratory for acrylamide, were also performed for glycidamide to determine its mutagenic properties and to compare responses. Results of these studies are consistent with the proposal that in vivo conversion to glycidamide is responsible for the mutagenicity of acrylamide in male mice.

Animals↗

Acrylamide in the Baylis-Hillman reaction: expanded reaction scope and the unexpected superiority of DABCO over more basic tertiary amine catalysts.

DMAP, DBU, and quinuclidine efficiently promote novel hydroalkoxylation reactions of acrylamide in primary alcohol solvents. DABCO is a comparatively poor hydroalkoxylation promoter and can effect clean, selective Baylis-Hillman reactions between acrylamide and aldehydes in alcoholic/aqueous media in which more basic nucleophilic catalysts promote hydroalkoxylation preferentially. Optimization of the reaction conditions has allowed acrylamide to be reacted with a range of aromatic aldehydes in moderate to excellent yields, including the first examples involving deactivated, electron-rich substrates such as p-tolualdehyde and o-anisaldehyde.

Journal Article↗

Alterations in nerve and muscle compound action potentials after acute acrylamide administration.

The early deficits of neurotoxicity induced by acrylamide were examined in rats by comparing nerve and muscle action potentials before and 24 hr after exposure to acrylamide (25, 50 or 100 mg/kg). No changes were seen in the nerve action potential amplitude or duration. The 25 mg/kg dose produced a more variable nerve conduction velocity. There was also a significant broadening of the muscle compound action potential. Neither of these effects were seen in the fasted controls. However, the lengthening of the relative refractory period of the muscle action potential was highly correlated with losses in body weight in the treatment groups and was identical to changes seen in control animals which were fasted for 24 hr. The slowed conduction of the muscle action potential may be a precursor of the nerve terminal damage which results from chronic exposure. Changes in the muscle refractory period, on the other hand, appear to be secondary to the loss in body weight which accompanies acrylamide administration.

Action Potentials↗

Micronuclei and developmental abnormalities in 4-day mouse embryos after paternal treatment with acrylamide.

The developmental consequences of paternal exposure to acrylamide (50 mg/kg i.p. for 5 days) were assessed in preimplantation embryos. There was a significant increase in the proportion of morphologically abnormal embryos after postmeiotic treatment during spermatogenesis (88.7% vs. 14.8% in control). Abnormal embryos had an average of 1.8 +/- 3.5 cells and > 80% had at least one fragmented nucleus. In addition, morphologically normal embryos were significantly delayed (34.3 +/- 12.8 cells per embryo vs. 57.6 +/- 15.7 in control, P < 0.001). Acrylamide caused 10- and 20-fold increases in frequencies of cells with micronuclei (MN) in morphologically normal and abnormal embryos, respectively (41 and 93 MN per 1,000 cells). Both centromere-negative (MN-) and centromere-positive (MN+) were induced. Nuclei of abnormal embryos were significantly larger (900 microm2 vs. 250 microm2) than controls. In addition, MN of abnormal embryos were larger than those of normal embryos (21.2 microm2 vs. 6.5 microm2, P < 0.01). Among control embryos, MN+ were significantly larger than MN- (P < 0.05). These findings suggest that the preimplantation embryo is a sensitive indicator of paternally transmitted effects on early development. Multiple mechanisms appear to be involved, including cytogenetic damage, proliferation arrest/delay, and fertilization failure. Future studies are needed to establish how induced cytological defects in preimplantation embryos contribute to birth defects and other postimplantation abnormalities.

Abnormalities, Drug-Induced↗

Effect of protein application mode and acrylamide concentration on the resolution of protein spots separated by two-dimensional gel electrophoresis.

Two-dimensional gel electrophoresis separates large numbers of proteins in two steps on the basis of differences in their pIs and molecular masses. The separation is usually performed on immobilized pH gradient strips, followed by gradient polyacrylamide gels separating proteins with molecular masses between 5-200 kDa. For the first-dimensional separation the protein samples are usually applied near one end of the strip. Using total soluble protein extracts of the bacterium Haemophilus influenzae, we found that simultaneous sample application at both the basic and the acidic ends of the strip resulted in detection of more and stronger protein spots in comparison with sample application at one end only. Because many proteins of an organism have similar pI and Mr values, an overlapping of protein spots is frequently observed in the second-dimensional separation. The soluble protein fraction of H. influenzae was further separated on gels of constant acrylamide concentration between 7.5% and 15.0%. We found that for proteins of molecular mass within certain ranges, the gels of homogeneous acrylamide concentration provided more efficient spot separation than the gradient gels. The observed improvements in spot resolution may be useful in the characterization of proteins from other organisms or cell lines.

Acrylamide↗

The effects of acrylamide on mouse germ-line and somatic cell chromosomes.

The industrial chemical acrylamide is suspected to induce potentially heritable genetic damage. While several studies in rodents have indicated that this substance can damage spermiogenic cells, resulting in dominant lethals and heritable translocations, cytogenetic assessments of premeiotic and meiotic cells after exposure have produced equivocal results. In the present study, various cytogenetic endpoints in both somatic and germ-line cells from acrylamide-treated mice were evaluated. Sister chromatid exchanges and micronuclei, but not chromosome aberrations, were induced in spleen cells; synaptonemal complex irregularities (asynapsis), but not chromosome aberrations, were induced in germ cells.

Acrylamide↗

Acrylamide: induction of heritable translocation in male mice.

Acrylamide (AA), known to induce dominant lethals in male rodents, was studied in the mouse heritable translocation test by using intraperitoneal injections on 5 consecutive days. Matings on days 7-10 following the last injection yielded a high frequency of translocation carriers in the F1 male population, which demonstrated that acrylamide is an effective inducer of translocations in postmeiotic germ cells. As an inducer of both dominant lethals and heritable translocations in late spermatids and early spermatozoa, AA is similar to alkylating agents such as ethylmethanesulfonate and ethylene oxide. However, AA's chemical structure, the nature of adducts formed with DNA, and it lack of mutagenicity in bacteria suggest a different mechanism as the basis for AA's germ cell mutagenicity.

Acrylamide↗