Quantification Of Heterocyclic Amines Research Articles

Quantification of hetero-cyclic amines from different categories of braised beef by optimized UPLC-TQ-XS/ESI method.

This research work has developed and optimized a sensitive analytical method for separation and quantification of heterocyclic amines (HCAs) mainly including PhIP, Harman, Norharman, IQ, MeIQ, AαC, MeAαC and Trp-P-2 by optimizing UPLC-TQ-XS using electrospray ionization source (ESI+) on ACQUITY UPLC® BEH C18 column in <7min, from braised beef sample matrix. Meanwhile, modified HCAs extraction by modifying QuEChERS (quick, easy, cheap, efficient, rugged and safe) technique and revisited with solid phase extraction (SPE) for HCAs purification, instead using traditional QuEChERS salts. Moreover, optimized pH conditions of HCA extracts before purification, for better extraction recoveries. Furthermore, this method was validated in terms of method validation parameters. Lastly, simulation of real braised beef model provided the minimum formation of HCAs by optimizing cooking parameters and precursors in a cooking system. Therefore, this method could be applied simultaneously on braised beef matrix either marketed or home cooked for HCAs analysis.

Analysis, Formation and Inhibition of Heterocyclic Amines in Foods: An Overview

Heterocyclic amines (HAs), an important class of toxicological compounds, can be formed through heating of four precursors, amino acids, creatinine, creatine and sugars. Numerous studies have shown that most HAs are mutagenic and carcinogenic, and the safety of HAs-containing foods has become a major concern for the public. This paper deals with an overview of analysis, formation and inhibition of HAs in foods. Analysis of HAs from foods is routinely conducted by protein precipitation, followed by acid-base solvent partition, purification with column chromatography and identification and quantification by high-performance liquid chromatography (HPLC), or gas chromatography-mass spectrometry (GC-MS). However, most of these methods result in low recovery. With HPLC, all 16 HAs can be simultaneously separated by a binary or ternary gradient solvent system and detected by UV. Using fluorescence detection, only nine HAs can be detected. For GC-MS, only a number of HAs are separated and identified by employing a temperature programming method. GC-MS provides rapid identification and quantification of HAs in foods, however, this technique requires a time-consuming derivatization and the number of HAs separation is limited. Also, some HAs may degrade and result in peak tailing. The application of HPLC with photodiode-array detection (HPLC-DAD) can not only permit rapid identification and quantification of HAs, but also enhance sensitivity and selectivity. Broiling and frying are two major processing methods causing the formation of high amount of HAs. The formation of HAs in foods during cooking can be inhibited by incorporating additives such as antioxidants and amino acids. However, the results have been controversial. Further research is necessary to study the effect of various additives on inhibition of HAs formation in foods.

Rapid determination of heterocyclic amines in ruminant meats using accelerated solvent extraction and ultra-high performance liquid chromatograph–mass spectrometry

Cooking techniques such as grilling confer several benefits to meat during food preparation including improved palatability, digestibility, preservation, and safety, as well as enhancing the sensory characteristics and net nutritional gain. However, grilling can lead to the formation of harmful compounds such heterocyclic amines (HCAs). HCAs are potent carcinogenic and mutagenic nitrogen containing compounds produced during certain cooking conditions of protein rich foods. Dietary intake of HCAs is associated with increased risk factors for cancers in humans. As such, there is overwhelming interest in identifying improved methods for rapid and accurate determination of heterocyclic amines in food matrices that is sensitive and avoids exhaustive sample preparation steps. Herein, we describe an approach that involves first extracting HCAs by pressurized accelerated solvent extractor using methanol as solvent, followed by addition of internal standard and quantification of HCAs by ultra-high performance liquid chromatography-high resolution accurate mass spectrometric detection (UHPLC-HRAMS). This method is fast, accurate, reproducible and does not require exhaustive sample pre-treatments prior to UHPLC-HRAMS analysis compared to existing/traditional methods for HCA analysis.•The method is automated, fast and uses tunable pressurized liquid extractor to selectively extract HCAs•Method does not require exhaustive cleanup and preconcentration steps prior to UHPLC/HRAMS analysis of HCAs•Validation showed method to be accurate, precise, and useful for routine multi-sample HCA analyses

Optimized HPLC Method for Analysis of Polar and Nonpolar Heterocyclic Amines in Cooked Meat Products

A simple, precise, and specific column high-performance liquid chromatographic (HPLC) method with UV absorption diode array and fluorescence detection has been developed by optimizing a previously described method for the simultaneous quantification of 15 polar and nonpolar heterocyclic amines (HAs) in fried meat products. The HPLC determination could be improved due to the application of a silica-based reversed-phase column with octadecyl groups (TSK-gel Super ODS) and a particle size of 2 microm. The separation of HAs in the complex meat matrix was performed with a 21 min mobile phase gradient. The method was validated for instrumental precision, repeatability, and selectivity and compared with a previously published method. After liquid adsorption of the basic sample mixture on diatomaceous earth, HAs were extracted with ethyl acetate. For cleanup, solid-phase extraction (silica propylsulfonic acid and octadecyl cartridges) and different washing steps were applied. Both nonpolar and polar HAs were determined in one fraction. The calibration curves of all HAs were linear for the applied detection system (correlation coefficient = 0.990-0.995). The recoveries, with the exception of 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2), were between 42 and 98% from meat samples spiked in a range of 1.5 to 3.3 ng/g for fluorescence-active and 4.3 to 8 ng/g for UV-active HAs. For quantification of HAs, the standard addition method was used for adjustment of different characteristics of HAs in the extraction. In fried meat samples (chicken breast and beef patties), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MelQx), 2-amino-3,4,8-trimethylimidazo[4,5-f] quinoxaline(4,8-DiMelQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), norharmane, and harmane were found in a concentration range of 0.02 to 14.3 ng/g.

Quantification of heterocyclic amines from thermally processed meats selected from a small‐scale population‐based study

Heterocyclic amines (HAs) are potent mutagens that form at high temperatures in cooked, protein-rich food. Due to their frequent intake, these compounds are considered a risk factor for human cancer. Cooking conditions and eating habits strongly influence the level of HA exposure. Thus, it is difficult to assess the intake of HAs in a large population. Food-frequency questionnaires (FFQs), designed to provide data on parameters that affect HA formation, were used to survey a small population (459 persons) from Barcelona (NE Spain). Subsequently, the most-consumed food items named were cooked according to the preferences of the population surveyed and analyzed for HAs using SPE and LC-MS/MS. In the population studied, the estimated intake via consumption of 13 meat dishes was 285.6 ng of mutagenic HAs per capita and day. PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) was the HA to which the population was most exposed, mainly from fried chicken and griddled beef. When the co-mutagens norharman and harman are included, the mean daily intake of HAs rises to 475.6 ng per capita and day. A novel putative DMIP regioisomer was detected in the cooked meats, which was analyzed in the present study by multistage MS.

Kinetics of Formation of Polar Heterocyclic Amines in a Meat Model System

ABSTRACTA model system was used to examine kinetics of formation of polar heterocyclic amines by heating the precursors creatinine, glucose and amino acids in proportions similar to those in bovine meat but at higher concentrations. Formation of heterocyclic amines was studied between 150 and 225°C for 0.5–120 min, depending on temperature. Heated samples were subjected to solid phase extraction and HPLC analysis, with photodiode array detection for identification and quantification of heterocyclic amines. IQx, MeIQx, 7,8‐DiMeIQx, 4,8‐DiMeIQx, PhIP, harman and norharman were identified. A first‐order reaction model and the Eyring equation were fitted to the formation of polar heterocyclic amines to obtain rate constants and their temperature dependence.

Quantification of heterocyclic amines in cooked foods

Quantification of heterocyclic amines in cooked foods