Heat-processed Foods Research Articles

Furosine, a Maillard Reaction Product, Triggers Necroptosis in Hepatocytes by Regulating the RIPK1/RIPK3/MLKL Pathway.

As one of the typical Maillard reaction products, furosine has been widely reported in a variety of heat-processed food. Though furosine was shown to be toxic on organs, its toxicity mechanism is still unclear. The present study aimed to investigate the toxicity mechanism of furosine in liver tissue. An intragastric gavage mice model (42-day administration, 0.1/0.25/0.5 g/kg of furosine per day) and a mice primary hepatocyte model were employed to investigate the toxicity mechanism of furosine on mice liver tissue. A metabonomics analysis of mice liver, serum, and red blood cells (RBC) was performed. The special metabolic mediator of furosine, lysophosphatidylcholine 18:0 (LPC (18:0)) was identified. Then, the effect of the upstream gene phospholipase A2 gamma (PLA2-3) on LPC (18:0), as well as the effect of furosine (100 mg/L) on the receptor-interacting serine/threonine-protein kinase (RIPK)1/RIPK3/mixed lineage kinase domain-like protein (MLKL) pathway and inflammatory factors, was determined in liver tissue and primary hepatocytes. PLA2-3 was found to regulate the level of LPC (18:0) and activate the expression of RIPK1, RIPK3, P-MLKL, and of the inflammatory factors including tumor necrosis factor α (TNF-α) and interleukin (IL-1β), both in liver tissue and in primary hepatocytes. Upon treatment with furosine, the upstream sensor PLA2-3 activated the RIPK1/RIPK3/MLKL necroptosis pathway and caused inflammation by regulating the expression of LPC (18:0), which further caused liver damage.

Loop-mediated isothermal amplification (LAMP) approach for detection of heat-resistant Talaromyces flavus species

Talaromyces flavus is a soilborne fungus that can contaminate fruits. It constitutes serious influence on heat-processed food spoilage, as T. flavus belongs to the heat-resistant fungi group, which are able to survive the pasteurization process. Moreover T. flavus has been reported to be capable of mycotoxigenicity, therefore they have a serious threat to human health. To maintain the safety of food production, sensitive method for T. flavus detection was developed. The loop mediated amplification, abbreviated LAMP, reactions were designed as specific for detection of DNA replication licensing factor gene of T. flavus. The specificity of assay was confirmed by use of 5 T. flavus strains and 35 other fungal isolates. The achieved limit of detection was 1fg of T. flavus genomic DNA and 64 ascospores in 1 g of strawberry fruits or soil samples.

Fluorescence quenching biosensor for acrylamide detection in food products based on double-stranded DNA and gold nanoparticles

To protect human health from the side effects of acrylamide in heat-processed food samples, simple analytical approaches are highly desired to determine low concentrations of acrylamide. In this study, a simple, rapid and accurate fluorescent sensor was developed for detection of acrylamide based on gold nanoparticles (AuNPs) and FAM-labeled double-stranded DNA (FAM-dsDNA). The sensing method was developed in a way to produce a remarkable fluorescence intensity difference in the absence and presence of acrylamide. In the presence of acrylamide, the single-stranded DNA (ssDNA) and acrylamide adduct is formed. So that, FAM-labeled complementary strand DNA (FAM-csDNA) is free in the environment and adsorbed on the surface of AuNPs and as a result, the FAM is quenched by the AuNPs. Under optimized conditions, the presented fluorescent analytical approach showed high selectivity toward acrylamide with a wide linear response, 1 × 10−7 M–0.05 M, and a limit of detection (LOD) of 1 × 10−8 M for acrylamide. This new method indicated excellent analytical performance for acrylamide detection in potato fries water extract samples with LOD of 0.5 × 10−6 M.

Toxicology studies of furosine in vitro/in vivo and exploration of the related mechanism

AimFurosine is one of the Maillard reaction products (MRPs) and is found in a variety of heat-processed food. Yet its toxicity is still unclear. The present study was designed to assess furosine toxicity in cell models and in CD-1 mice, respectively. MethodsIn vitro, the effects of furosine on the cell viability, cell cycle and apoptosis (Hek293, HepG2, SK-N-SH and Caco2) were detected and evaluated, sensitive cell lines and proper dosage of furosine for further animal experiment were determined, and the mechanisms of toxicity were explored. In vivo, the acute toxicity studieswere performed, organ index, hematology parameters, functions of liver/kidney and pathological changes were detected and the target organs were uncovered. ResultsHek293 cells and HepG2 cells were themost sensitive to furosine with respect to cytotoxicity and apoptosis. Furosine inhibited mice weight gain, and affected the functions of liver and kidney. ConclusionsFurosine posed toxic effects on mice liver and kidney, suggested thatthey were the target organs for furosine toxicity. This study for the first time provides evidence that high dosages of furosine pose adverse biological effects on the health of animals through induction of cell apoptosis and activation of inflammatory necrosis response.

Development of a qPCR assay for the detection of heat-resistant Talaromyces flavus

Heat-resistant fungi of the species Talaromyces flavus, which inhabits soil and can contaminate fruits, constitutes significant impact on spoilage of heat-processed food. T. flavus possess the ability to produce numerous mycotoxins and is able to survive the process of pasteurization what makes it a treat to food industry. Up to date there is no rapid and reliable method to detect and identify T. flavus. Therefore in this study, a sensitive method for detecting T. flavus was developed. The primers (Tf1_F/R) specific to detection of DNA replication licensing factor gene of T. flavus were designed. With this set of primers, a qPCR reaction with SybrGreen detection was developed. The specificity of assay with use of 5 T. flavus strains and 35 other fungal isolates was tested. The detection threshold was 200 fg of T. flavus genomic DNA. The developed method was able to detect 640 ascospores in 1 g of strawberry fruits and soil samples.

Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed.

Food adulteration and feed contamination are significant issues in the food/feed industry, especially for meat products. Reliable techniques are needed to monitor these issues. Droplet Digital PCR (ddPCR) assays were developed and evaluated for detection and quantification of bovine, porcine, chicken and turkey DNA in food and feed samples. The ddPCR methods were designed based on mitochondrial DNA sequences and integrated with an artificial recombinant plasmid DNA to control variabilities in PCR procedures. The specificity of the ddPCR assays was confirmed by testing both target species and additional 18 non-target species. Linear regression established a detection range between 79 and 33200 copies of the target molecule from 0.26 to 176 pg of fresh animal tissue DNA with a coefficient of determination (R2) of 0.997–0.999. The quantification ranges of the methods for testing fortified heat-processed food and feed samples were 0.05–3.0% (wt/wt) for the bovine and turkey targets, and 0.01–1.0% (wt/wt) for pork and chicken targets. Our methods demonstrated acceptable repeatability and reproducibility for the analytical process for food and feed samples. Internal validation of the PCR process was monitored using a control chart for 74 consecutive ddPCR runs for quantifying bovine DNA. A matrix effect was observed while establishing calibration curves with the matrix type under testing, and the inclusion of an internal control in DNA extraction provides a useful means to overcome this effect. DNA degradation caused by heating, sonication or Taq I restriction enzyme digestion was found to reduce ddPCR readings by as much as 4.5 fold. The results illustrated the applicability of the methods to quantify meat species in food and feed samples without the need for a standard curve, and to potentially support enforcement activities for food authentication and feed control. Standard reference materials matching typical manufacturing processes are needed for future validation of ddPCR assays for absolute quantification of meat species.

Ohmic heating: A promising technology to reduce furan formation in sterilized vegetable and vegetable/meat baby foods

Abstract Occurrence of furan, classified as carcinogen 2B by the International Agency for Research on Cancer, in heat-processed foods, especially sterilized baby foods, is of a health concern. On this account, innovative processing practices ensuring microbial safety, acceptable sensorial features, and, at the same time, minimizing furan formation have to be searched. In this study, the potential of ohmic heating to mitigate furan formation was demonstrated. Compared to conventional retort sterilization, significant mitigation (70–90%) of furan was achieved, assumingly due to reduced degradation of furan precursor under faster terms heating conditions. In purees containing meat, approx. two times less furan was formed, regardless of the processing technology. In addition to furan, also other headspace volatiles were measured and statistically evaluated. Compounds originated through fatty acids oxidation and Maillard reaction products were more abundant in conventionally sterilized samples compared to those treated by ohmic heating. Industrial relevance Ohmic heating is an emerging technology being employed in the field of food processing which applies a direct electric current to food products, providing rapid and uniform heating throughout the product. Shorter heating times used in ohmic heating, as compared to conventional retort sterilization, reduce potential losses of valuable nutrients and as well as reduce the formation of undesirable processing contaminants, in particular furan. The presented work examines furan concentrations across various stages of baby food production, in order to compare ohmic heating and retort sterilization processes. Volatile compound fingerprints for baby food purees processed via both sterilization methods, both prior and post sterilization were assessed. The results presented in this work are of high potential interest to the baby food industry to reduce both heat induced chemical changes and exposure of infants and babies to hazardous processing contaminants such as furan.

Association between habitual dietary and lifestyle behaviours and skin autofluorescence (SAF), a marker of tissue accumulation of advanced glycation endproducts (AGEs), in healthy adults.

Advanced glycation endproducts (AGEs) are produced endogenously and also enter the body during the consumption of AGEs present in heat-processed food. It is unknown whether AGEs of dietary origin accumulate within the body of healthy individuals. AGEs can deposit within skin tissue long-term by crosslinking extracellular matrix proteins. The fluorescent nature of many AGEs enables their detection within the skin by non-invasively measuring skin autofluorescence (SAF). This study aimed to identify habitual dietary and lifestyle behaviours cross-sectionally associated with SAF in an adult population sample. 251 Healthy adult volunteers completed validated food frequency and physical activity questionnaires. Waist circumference, BMI, blood pressure and blood glucose was also measured. SAF was measured using an AGE Reader. Significant positive correlations were found between SAF and chronological age (r=0.63, P<0.001), waist circumference (r=0.28, P<0.01), body weight (r=0.24, P<0.05), BMI (r=0.23, P<0.05) and consumption of meat and meat products (r=0.22, P<0.05). A negative correlation was found between SAF and cereal consumption (r=-0.21, P<0.05). Cigarette smokers also had a significantly higher SAF than non-smokers (2.4 vs 2.0U, P<0.05). Regression analysis identified age, cigarette smoking, waist circumference and intake of meat products as significant predictors of SAF. The regression model explained 48% of the variation in SAF. Age, cigarette smoking, waist circumference and dietary consumption of meat/meat products were positively associated with SAF in this sample. Further research is required to determine whether frequent consumption of foods containing large quantities of dietary AGEs contribute to pathological disease processes in healthy individuals.

Process-induced formation of imidazoles in selected foods

The presence of 4-methylimidazole (4-MEI), 2-methylimidazole (2-MEI) and 2-acetyl-4-tetrahydroxybutylimidazole (THI) in some foods may result from the usage of caramel colorants E150c and E150d as food additives. This study demonstrates that alkylimidazoles are also byproducts formed from natural constituents in foods during thermal processes. A range of heat-processed foods that are known not to contain caramel colorants were analyzed by isotope dilution LC-MS/MS to determine the contamination levels. Highest 4-MEI concentrations (up to 466µg/kg) were observed in roasted barley, roasted malt and cocoa powders, with the concomitant presence of 2-MEI and/or THI in some cases, albeit at significantly lower levels. Low amounts of 4-MEI (<20µg/kg) were also detected in cereal-based foods such as breakfast cereals and bread toasted to a brown color (medium toasted). The occurrence of 4-MEI in certain processed foods is therefore not a reliable indicator of the presence of the additives E150c or E150d.

Relationship between HMF intake and SMF formation in vivo: An animal and human study.

5-Hydroxymethylfurfural (HMF) is a furanic compound produced in heat-processed foods by nonenzymatic browning reactions. HMF has been demonstrated to be hepato- and nephrotoxic in animals with a link to its metabolite 5-sulfooxymethylfurfural (SMF). To date little is known about either the formation of SMF from ingested HMF or the formation of DNA adducts in animals or human beings. To assess SMF in vivo formation, we first performed a study in mice treated with high/low doses of oral HMF. We found increased concentrations of SMF in plasma and DNA SMF-adducts in leukocytes, hepatic tissue, and kidneys by means of LC-MS/MS, but no spatial formation in such tissues was observed by MALDI-MS imaging technology due to low sensitivity. In a second experiment, we measured the exposure to HMF in a Spanish preadolescent population. We analyzed the concentration of HMF metabolites (plasma, urine) and measured, for the first time, the presence of SMF in plasma and DNA SMF-adducts in leukocytes. This study provides the first evidence that oral HMF is readily transformed into SMF in vivo, giving rise to the formation of DNA adducts in a direct relation with HMF intake, both in animals and human beings.

New insights into the molecular mechanisms of chemical carcinogenesis: In vivo adduction of histone H2B by a reactive metabolite of the chemical carcinogen furan

Furan is a rodent hepatocarcinogen ubiquitously found in the environment and heat-processed foods. Furan undergoes cytochrome P450 2E1-catalyzed bioactivation to cis-2-butene-1,4-dial (BDA), which has been shown to form an electrophilic conjugate (GSH-BDA) with glutathione. Both BDA and GSH-BDA yield covalent adducts with lysine residues in proteins. Dose- and time-dependent epigenetic histone alterations have been observed in furan-treated rats. While the covalent modification of histones by chemical carcinogens has long been proposed, histone-carcinogen adducts have eluded detection in vivo. In this study, we investigated if the covalent modification of histones by furan may occur in vivo prior to epigenetic histone alterations.Using a “bottom-up” methodology, involving the analysis of tryptic peptides by liquid chromatography – high resolution mass spectrometry, we obtained evidence for a cross-link between GSH-BDA and lysine 107 of histone H2B isolated from the livers of male F344 rats treated with tumorigenic doses of furan. This cross-link was detected at the shortest treatment period (90 days) in the lowest dose group (0.92mg/kg body weight/day), prior to the identification of epigenetic changes, and occurred at a lysine residue that is a target for epigenetic modifications and crucial for nucleosome stability. Our results represent the first unequivocal proof of the occurrence of carcinogen-modified histones in vivo and suggest that such modification happens at the initial stages of furan-induced carcinogenesis. This type of alteration may be general in scope, opening new insights into the mechanisms of chemical carcinogenesis/toxicity and new opportunities for the development of early compound-specific biomarkers of exposure.

Dummy-surface molecularly imprinted polymers on magnetic graphene oxide for rapid and selective quantification of acrylamide in heat-processed (including fried) foods.

Novel nano-sized dummy-surface molecularly imprinted polymers (DSMIPs) on a magnetic graphene oxide (GO-Fe3O4) surface were developed as substrates, using propionamide as a dummy template molecule for the selective recognition and rapid pre-concentration and removal of acrylamide (AM) from food samples. These products showed rapid kinetics, high binding capacity (adsorption at 3.68mg·g-1), and selectivity (imprinting factor α 2.83); the adsorption processes followed the Langmuir-Freundlich isotherm and pseudo-second-order kinetic models. Excellent recognition selectivity toward acrylamide was achieved compared to structural analogs, such as propionic and acrylic acids (selectivity factor β 2.33, and 2.20, respectively). Moreover, DSMIPs-GO-Fe3O4 was used to quantify acrylamide in food samples, yielding satisfactory recovery (86.7-94.3%) and low relative standard deviation (<4.85%). Thus, our DSMIPs-GO-Fe3O4-based procedure was demonstrated to be a convenient and practical method for the separation, enrichment, and removal of acrylamide from food samples.

A novel reference real-time PCR assay for the relative quantification of (game) meat species in raw and heat-processed food

In order to increase their profit, food producers may be tempted to replace expensive meat species by cheaper ones. Due to their selectivity and sensitivity, PCR based methods are frequently applied to identify and quantify meat species to detect food adulteration. Quantification of meat species in highly processed food products is, however, a difficult task. When we used a previously published reference system to relatively quantify the deer content in heat treated foodstuffs, we obtained recoveries that were substantially >100%. In the present study we aimed to improve the applicability of the reference system, targeting a 97 bp fragment of the myostatin gene, to heat treated products by reducing the length of the target sequence. The novel reference system, targeting a 70 bp fragment of the myostatin gene, was found to amplify the target region in 27 mammals and poultry species. Meat mixtures as well as raw and heat treated model sausages were analysed to demonstrate the applicability of the novel reference system for relatively quantifying the game meat content in processed food products. With the novel reference system, the bias introduced by heat treatment was largely eliminated.

Simultaneous determination of furan and 2-alkylfurans in heat-processed foods by automated static headspace gas chromatography-mass spectrometry

In this study, the levels of furan and 2-alkylfurans (2-methylfuran, 2-ethylfuran, 2, 5-dimethylfuran, 2-propylfuran, 2-butylfuran, 2-pentylfuran) were determined in a variety of foodstuffs by static headspace gas chromatography and mass spectrometry. The headspace operating conditions and the effects of food matrix on the responses of analytes were investigated. Besides, the co-elution of 2-ethylfuran/2, 5-dimethylfuran was processed using the automated deconvolution software for the correct identification and quantification of 2-ethylfuran and 2,5-dimethylfuran in food samples. The external matrix matched standard calibration method was chosen to correct the matrix effects for the analysis of furan and 2-alkylfurans in a total of 126 food samples. Results showed that furan and 2-alkylfurans were detected in 70.6% of food samples with total concentrations ranging from 0.5 to 789.4 ng g−1.

A simultaneous derivatization of 3-monochloropropanediol and 1,3-dichloropropane with hexamethyldisilazane–trimethylsilyl trifluoromethanesulfonate at room temperature for efficient analysis of food sample analysis

This paper reports the application of hexamethyldisilazane–trimethylsilyl trifluoromethanesulfonate (HMDS–TMSOTf) for the simultaneous silylation of 3-monochloro-1,2-propanediol (3-MCPD) and 1,3-dicholoropropanol (1,3-DCP) in solid and liquid food samples. 3-MCPD and 1,3-DCP are chloropropanols that have been established as Group 2B carcinogens in clinical testing. They can be found in heat-processed food, especially when an extended high-temperature treatment is required. However, the current AOAC detection method is time-consuming and expensive. Thus, HMDS–TMSOTf was used in this study to provide a safer, and cost-effective alternative to the HFBI method. Three important steps are involved in the quantification of 3-MCPD and 1,3-DCP: extraction, derivatization and quantification. The optimization of the derivatization process, which involved focusing on the catalyst volume, derivatization temperature, and derivatization time was performed based on the findings obtained from both the Box–Behnken modeling and a real experimental set up. With the optimized conditions, the newly developed method was used for actual food sample quantification and the results were compared with those obtained via the standard AOAC method. The developed method required less samples and reagents but it could be used to achieve lower limits of quantification (0.0043mgL−1 for 1,3-DCP and 0.0011mgL−1 for 3-MCPD) and detection (0.0028mgL−1 for 1,3-DCP and 0.0008mgL−1 for 3-MCPD). All the detected concentrations are below the maximum tolerable limit of 0.02mgL−1. The percentage of recovery obtained from food sample analysis was between 83% and 96%. The new procedure was validated with the AOAC method and showed a comparable performance. The HMDS–TMSOTf derivatization strategy is capable of simultaneously derivatizing 1,3-DCP and 3-MCPD at room temperature, and it also serves as a rapid, sensitive, and accurate analytical method for food samples analysis.

Direct determination of acrylamide in food by gas chromatography with nitrogen chemiluminescence detection

A method of gas chromatography with nitrogen chemiluminescence detection and using standard addition is described for the determination of acrylamide in heat-processed foods. Using a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation method removes the acrylamide precursors completely, and the risk of overestimating acrylamide concentration due to additional analyte formation in the hot gas chromatograph inlet is also avoided. Sample preparation is rapid and inexpensive. A Deans switch device is utilized to heart-cut acrylamide and to prevent interferences from the solvent and matrix from reaching the detector. The pre-column is backflushed at high temperature to maintain a clean baseline and shorten the cycle time compared to baking out the column. Quantitation using standard addition is employed for compensation of potential variability in the acrylamide extraction efficiency in acetonitrile. The limit of detection and the limit of the quantification obtained for this method are 27 and 81 μg/kg, respectively, in food samples (equivalent to 3.5 and 10.6 μg/L in acetonitrile, respectively), and the linear range is 76-9697 μg/kg in food samples (equivalent to 10-1280 μg/L in acetonitrile) with an R(2) value of 0.9999.

The kinetics of the inhibition of acrylamide by glycine in potato model systems.

Acrylamide (AA) is a potential carcinogen which widely exists in heat-processed foods. The addition of glycine (Gly) has been shown to reduce the formation of AA. The objective of this work was to investigate the kinetics of the inhibition of AA by Gly in both asparagine (Asn)/glucose (Glc) and Asn/Glc/Gly potato model systems during heating at 160 °C, 180 °C, and 200 °C. The simplified two consecutive first-order kinetic model fitted well to the changes of AA in both systems. No significant difference in rate constant (kF) and apparent activation energy (EaF) was observed for AA formation between the two systems (P > 0.05). Whereas EaE and only kE at 200 °C for AA elimination in the Asn/Glc/Gly system was significantly higher than Asn/Glc system (P < 0.05). The elimination reaction between Gly and AA was confirmed by the identification of their major reaction product 2-((3-amino-3-oxopropyl)amino)acetic acid in the Asn/Glc/(15) N-Gly system. The reduction of AA by Gly is predominantly attributed to the elimination reaction between Gly and AA.

Potential protective effect of L-cysteine against the toxicity of acrylamide and furan in exposed Xenopus laevis embryos: an interaction study.

The embryo toxicities of two food-processing-induced toxic compounds, acrylamide and furan, with and without added L-cysteine were examined individually and in mixtures using the frog embryo teratogenesis assay-Xenopus (FETAX). The following measures of developmental toxicity were used: (a) 96 h LC50, the median concentration causing 50% embryo lethality; (b) 96 h EC50, the median concentration causing 50% malformations of the surviving embryos; and (c) teratogenic index (96 h LC50/96 h EC50), an estimate of teratogenic risk. Calculations of toxic units (TU) were used to assess possible antagonism, synergism, or response addition of several mixtures. The evaluated compounds demonstrated counterintuitive effects. Furan had lower than expected toxicity in Xenopus embryos and, unlike acrylamide, does not seem to be teratogenic. However, the short duration of the tests may not show the full effects of furan if it is truly primarily genotoxic and carcinogenic. L-Cysteine showed unexpected properties in the delay of hatching of the embryos. The results from the interaction studies between combination of two or three components (acrylamide plus L-cysteine; furan plus L-cysteine; acrylamide plus furan; acrylamide plus furan and L-cysteine) show that furan and acrylamide seem to have less than response addition at 1:1 toxic unit ratio in lethality. Acrylamide and L-cysteine show severe antagonism even at low 19 acrylamide/1 L-cysteine TU ratios. Data from the mixture of acrylamide, furan, and L-cysteine show a slight antagonism, less than would have been expected from binary mixture exposures. Bioalkylation mechanisms and their prevention are discussed. There is a need to study the toxicological properties of mixtures of acrylamide and furan concurrently formed in heat-processed food.

Identification of Puffer Fish of the Genus Lagocephalus: L. lunaris, L. spadiceus and L. inermis, Using Multiplex PCR

A multiplex PCR assay was developed to identify widespread poison-containing puffer fishes of the genus Lagocephalus: L. lunaris, L. spadiceus and L. inermis, using a novel KUGEN_ILSspec primer set which is specific for 12S ribosomal RNA (12S rRNA) and Cytochrome b (Cyt b) genes. KUGEN_ILSspec set is composed of fish positive-amplified primers that produced a 289-bp fragment while L. lunaris-, L. spadiceus- and L. inermis-specific primers produced 123-, 196- and 493-bp fragments, respectively. The sensitivity of this primer set was found to be high as it was capable of amplifying targeted DNA at concentrations as low as 1 ng/μL. Moreover, the primer set was shown to amplify intact DNA and species-specific fragments from heat-processed food. Consequently, multiplex PCR technique in combination with KUGEN_ILSspec primer set could offer an effective measure for detecting poisonous puffer fish contamination in both fresh and processed fish products, which will greatly aid in public health control and law enforcing endeavors.

Pottenger's cats revisited: a classic example of a nutritional deficiency of taurine (820.4)

Objective. From 1932 to 1942, Dr. Francis Marion Pottenger Jr. conducted a group of experiments to determine the effects of heat‐processed food on cats. Cats fed an all‐raw diet were healthy while cats fed the cooked meat diet developed various health problems. At the time of Pottenger's studies the heat labile amino acid taurine had not yet been identified as essential for cats. This work shows that the deficiencies Pottenger identified in cats correspond with those of a taurine deficiency and are the direct result of the lack of taurine in the feline diet.Methods. Pub Med and EBSCO Host were utilized to obtain journal articles. The Pottenger papers were obtained from the Price‐Pottenger Nutritional Foundation. The physiological effects of a cooked diet described by Pottenger in his papers were compared with data from recent studies on taurine deficiency in the feline diet.Results. Pottenger’s main observations of near and far sightedness, cardiac leasions, increased stillbirth, low birthweight, poor kitten survival, and developmental abnormalities each correspond to published descriptions of taurine deficiency in cats with multiple references for each described condition.Conclusions. Taurine deficiency is a strong explanation for the symptoms observed by Pottenger in his cat studies. Pottenger’s own conclusion that there was an ‘as yet unidentified, heat‐labile protein factor’ is realized in taurine.