Fast protein and metabolite liquid chromatography (FPLMC) was introduced years ago to enable the easy separation of high-molecular compounds such as proteins from small molecules and the identification of the low-molecular substances. In this paper, the method is applied for the rapid evaluation of freshness and monitoring the aging of animal meat and fish. A novel chromatographic sensor was developed with a deep UV LED-based photometric detection unit (255–265 nm), an original flow cuvette and registration scheme; the processing of a chromatogram with the sensor takes approximately 15 min. Strict isochronism between the elution of ATP metabolites, mainly hypoxanthine (Hx) and inosine monophosphate (IMP), and the time of maturation of meat or fish, was discovered. A new freshness index H* = [Hx]/[IMP] was introduced, which is proportional to the instrumental delay time in the FPMLC chromatograms: the H* index < 0.5 indicates the presence of inosine monophosphate (IMP) and the high quality of the meat or fish. Reasonably strong correlations were revealed between data obtained by FPMLC and total volatile basic nitrogen TVB-N (for fish) or volatile fatty acids VFA (for meat) content. Moreover, putative nucleotide salvage and an increase in the concentration of IMP were observed in fish after heat treatment using the FPMLC sensor and NMR technique.

Point-of-care testing (POCT) devices are becoming increasingly popular in the medical community as an alternative to conventional laboratory testing, especially for home treatments or other forms of outpatient care. Multiple-use chemical sensors with minimal requirements for disposables are among the most practical and cost-effective POC diagnostic instruments, especially in managing chronic conditions. An affordable, simple, and easy-to-use optical sensor based on fast protein liquid chromatography with direct UV absorption detection was developed for the rapid determination of the total protein concentration in effluent peritoneal dialysate and for the assessment of protein losses in end-stage renal disease (ESRD) patients on constant ambulatory peritoneal dialysis (CAPD). The sensor employs non-disposable PD-10 desalting columns for the separation of molecules with different molecular weights and a deep UV LED (maximum at 285 nm) as a light source for optical detection. The analytic procedure is relatively simple, takes 10–15 min, and potentially can be performed by patients themselves or nursing staff without laboratory training. Preliminary clinical trials on a group of 23 patients on CAPD revealed a good concordance between the protein concentrations in dialysate samples measured with the sensor and an automated biochemical analyzer; the mean relative error was about 10%, which is comparable with routine clinical laboratory methods.

Dietary nucleotides and nucleosides, primarily inosine monophosphate (IMP) and the adenine nucleotide pool (ANP), are widely considered as essential nutrients responsible for multiple biological functions. Food prepared from meat and fish is the main source of these substances in the human diet, and it is extremely important to implement storage and processing techniques ensuring their maximum preservation and even accumulation during maturation or conditioning. In experiments with freshly refrigerated grass carp and defrosted Alaska pollock fillets it was discovered, initially using Fast Protein and Metabolites Liquid Chromatography and the ATP-bioluminescence test, and afterwards validated by NMR spectroscopy, that heat treatment identical to conventional culinary processing in aqueous or wet media at temperatures above 62 °C leads to nucleotide salvage (recovery) in aged fish. A significant increase in the concentration of IMP, and even an emergence of ANP substances, were reliably demonstrated in fish samples which had already partially or fully lost these components during prolonged storage due to the ATP breakdown metabolic reactions. Owing to this recovery, the nutritive value of ready-to-eat food can be higher than was initially evaluated in raw products before heat treatment: an effect that should certainly be considered in practical nutrition. Moreover, it is necessary to reconsider the widely acknowledged system of indices of freshness based on nucleotides and nucleosides elaborated a long time ago for raw meat and fish products.

A novel simple optical sensor based on fast protein liquid chromatography was developed and tested for monitoring end stage renal disease (ESRD) patients treated with continuous ambulatory peritoneal dialysis (CAPD). The device provides direct determination of proteins and lower molecular weight metabolites in effluent peritoneal dialysate using ultraviolet (UV) photometric detection at the wavelengths 285 nm or 260 nm with deep ultraviolet light-emitting diodes. The sensor was calibrated with bovine serum albumin and nucleotides standard solutions. Chromatograms of peritoneal dialysate samples taken from a group of 28 ESRD patients were processed and approximated by a set of split-Gaussian functions. All chromatograms show three overlapping peaks: the first one represents proteins; the other two peaks probably correspond to mid- and low molecular weight metabolites. Strong correlation was reveled between the area of the first peak and total protein concentration determined by a standard biochemical assay, this makes possible estimation of peritoneal protein loss with a reasonable precision less than 15%. The area of the second peak correlated with dialysate optical density at a wavelength 355–365 nm, associated with the UV absorption of advanced glycation end (AGE) products. The third peak correlated with the optical density of the eluate at a wavelength 255–265 nm, associated with the UV absorption of purines and pyrimidines. Thus, we demonstrated the possibility of estimation of proteins and lower molecular weight metabolites in effluent peritoneal dialysate with the compact and affordable chromatographic optical sensor.

It is commonly accepted that dietary nucleotides should be considered as essential nutrients originating mainly but not exclusively from meat and fish dishes. Most research in food science related to nutrition nucleotides is focused on raw products, while the effects of thermal processing of ready-to-eat food on nucleotide content are largely overlooked by the scientific community. The aim of this study is to investigate the impact of thermal processing and cold storage on the content of dietary nucleotides in freshly prepared and canned ready-to-eat meat and fish food. The concentrations of ATP, ADP, AMP, IMP, Ino, and Hx were determined using NMR, HPLC, FPMLC, and ATP bioluminescence analytical techniques; freshness indices K and K₁ and adenylate energy charge (AEC) values were estimated to assess the freshness status and confirm a newly unveiled phenomenon of the reappearance of adenylate nucleotides. It was found that in freshly prepared at 65 °C ≤ T ≤ +100 °C and canned food, the concentration of free nucleotides was in the range of 0.001–0.01 µmol/mL and remained unchanged for a long time during cold storage; the correct distribution of mole fractions of adenylates corresponding to 0 < AEC < 0.5 was observed compared to 0.2 < AEC < 1.0 in the original raw samples, with either a high or low content of residual adenylates. It could be assumed that heating at nonenzymatic temperatures T > 65 °C can rupture cell membranes and release residual intracell nucleotides in quite a meaningful concentration. These findings may lead to a conceptual change in the views on food preparation processes, taking into account the phenomenon of the free adenylates renaissance and AEC bioenergetics.

Fresh fish and poultry meat are in high demand on the market: poultry, mainly chicken, is the second most consumed and the most affordable meat product in the world. Fish consumption varies greatly across regions but, in some countries, seafood is the main source of abundant and affordable macro- and micronutrients. Meat and, especially, fish are highly perishable products; methods and equipment for rapid, objective, and reliable assessing the freshness of fish and meat are crucial for the food industry. Generally recognized reference techniques such as total volatile basic nitrogen (TVB-N), volatile fatty acids (VFA), high pressure liquid chromatography (HPLC), mass spectrometry, or nuclear magnetic resonance (NMR) spectroscopy are time-consuming and require expensive and complex equipment. We developed a novel chromatographic optical sensor with a deep UV LED photometric detection (255–265 nm) for rapid assessment of meat and fish freshness based on determination of the relative content of adenosine triphosphate (ATP) metabolites. The sensor has a simple and compact design, and relatively low cost; sample preparation and processing of a chromatogram takes less than 30 min. The sensor was tested on Amur (farmed freshwater fish) and rooster meat, obtained from a local farmer. The samples were kept refrigerated at +4 °C, measurements were taken daily during a 14 day period. All chromatograms show two peaks: proteins are responsible for the first one; the second broad post-protein band is formed due to the overlapping of individual peaks of ATP and its metabolites. As fish and poultry meat are stored, ATP is converted into metabolites with lower molecular weight, which is reflected in the chromatograms—the elution time for the second peak increases. It was shown that this time can be directly associated with the freshness status of a product. As expected, poultry meat showed better storage stability and freshness retention compared to Amur fish.