Tritium uptake in crops in the area with a high level of atmospheric tritium oxide in the territory of the former Semipalatinsk test site.

During the period from 2019 to 2021, a series of experiments were carried out to study the uptake of tritium by crops in an area heavily contaminated with atmospheric tritium oxide (HTO), at the former Semipalatinsk test site in Kazakhstan. A quantitative assessment is given of the tritium uptake by typical crops (lettuce, tomatoes, peppers and beans) cultivated all over Kazakhstan in the case of a short-term tritium oxide vapor exposure. The plant samples were collected during and after exposure and analyzed for the tritium concentration in two chemical forms: tissue-free water tritium (TFWT) and organically bound tritium (OBT). During the entire series of experiments, the tritium concentration in free water from leaves and ambient air was of the same order of magnitude. The tissue water tritium concentrations of stems and edible parts was 1 to 2 orders of magnitude lower than in the surrounding air. The average value of the TFWT/HTOatm ratio in the leaves and the edible part was (0.73±0.2) and (0.04±0.002), respectively. The organically-bound tritium concentration is 1-2 orders of magnitude lower than the tissue water tritium and ambient air concentrations. Under aerial tritium oxide uptake, the distribution of tritium in non-leafy crops was as follows: leaf-stem-fruit (in decreasing order). After exposure, a non-significant amount of tritium is firmly retained in plants for a long time. The tissue water tritium concentrations correlate closely with atmospheric tritium oxid (r = 0.76), correlate weakly with temperature (r = 0.43) and relative humidity (r = -0.43), and correlate moderately with solar radiation intensity (r = 0.56). There was no reliable correlation between the concentration of tritium in organic matter and in ambient air. The concentration of tritium in the free water of leaves is closely correlated with the concentration in the free water of the stems (r = 0.95) and fruits (r = 0.78). The organically-bound tritium concentration in leaves is closely correlated with the organically-bound tritium concentration in stems (r = 0.99) and fruits (r = 98). The results of the study should be considered when evaluating the impact of tritium oxide emissions on the population living near nuclear power.

The Development and Application of Tritium-Labeled Compounds in Biomedical Research.

With low background radiation, tritiate compounds exclusively emit intense beta particles without structural changes. This makes them a useful tool in the drug discovery arsenal. Thanks to the recent rapid progress in tritium chemistry, the preparation and analysis of tritium-labeled compounds are now much easier, simpler, and cheaper. Pharmacokinetics, autoradiography, and protein binding studies have been much more efficient with the employment of tritium-labeled compounds. This review provides a comprehensive overview of tritium-labeled compounds regarding their properties, synthesis strategies, and applications.

Dually Labeled Neurotensin NTS1R Ligands for Probing Radiochemical and Fluorescence-Based Binding Assays.

The determination of ligand-receptor binding affinities plays a key role in the development process of pharmaceuticals. While the classical radiochemical binding assay uses radioligands, fluorescence-based binding assays require fluorescent probes. Usually, radio- and fluorescence-labeled ligands are dissimilar in terms of structure and bioactivity, and can be used in either radiochemical or fluorescence-based assays. Aiming for a close comparison of both assay types, we synthesized tritiated fluorescent neurotensin receptor ligands ([3H]13, [3H]18) and their nontritiated analogues (13, 18). The labeled probes were studied in radiochemical and fluorescence-based (high-content imaging, flow cytometry, fluorescence anisotropy) binding assays. Equilibrium saturation binding yielded well-comparable ligand-receptor affinities, indicating that all these setups can be used for the screening of new drugs. In contrast, discrepancies were found in the kinetic behavior of the probes, which can be attributed to technical differences of the methods and require further studies with respect to the elucidation of the underlying mechanisms.

Validating the application of the revised ICRP's biokinetic models for organic14C and organically bound tritium to members of the public.

In 2016, the International Commission on Radiological Protection (ICRP) has revised the biokinetic models for carbon and tritium in Publication 134 to calculate the dose coefficients of these radionuclides for workers. The following publication for members of the public is now in the process of revising by the ICRP. According to the draft manuscript published for consultation in 2023, the same models will be adopted for members of the public, although the parameters in these models are not corroborated by the metabolic data of radionuclides in foods. Dose coefficients for adult were estimated using modified models developed in this study to validate the application of the revised ICRP models to members of the public. In the modified models, several parameters were replaced based on the metabolic data of these nuclides in foods and compartments of radio-insensitive tissues were introduced. For these estimations, we utilised an inhouse program for internal-dose calculation developed by the Japan Atomic Energy Agency. The estimated dose coefficient values for ingestion of organic14C and organically bound tritium (OBT) ranged from 3.2 × 10-11-7.6 × 10-11Sv Bq-1and from 3.5 × 10-11-5.4 × 10-11Sv Bq-1, respectively. We concluded that the dose coefficient value of 1.6 × 10-10Sv Bq-1obtained by the revised ICRP's carbon model was conservative for members of the public, while the value of 5.1 × 10-11Sv Bq-1for OBT was appropriate.

Radioecological characteristics of Siberian roe deer (Capreolus pygargus Pal., 1771) inhabiting locations of nuclear weapon tests.

This paper reports the activity concentrations of 137Cs, 90Sr, 239+240Pu, 241Am, and 3Н in the form of tritiated water (НТО) and organically bound tritium (ОBТ) in the tissues and organs of roe deer (Capreolus pygargus Pal., 1771) that inhabit the 'Degelen' test location of the Semipalatinsk Test Site. Tissues and organs were sampled from six deer by killing. The activity concentrations of specific radionuclides in the samples were measured using γ-, α-, and ß-spectrometry. The radionuclide activity concentrations in the tissues and organs showed considerable variation, for example, 0.6-170 Bq kg-1 for 137Cs and 0.3-2.8×103 Bq kg-1 for 90Sr. The activity concentrations of radionuclides in animal muscular tissue did not exceed permissible values for the meat of wild animals. The tissues and organs in the roe deer were arranged as follows in descending order of their ability to accumulate 137Cs and 90Sr: for 137Cs, muscular tissue-kidneys-lungs-spleen-heart-liver-bone tissue; for 90Sr, bone tissue-liver-lungs-muscular tissue-spleen-heart-kidneys. The activity concentrations of 241Am and 239+240Pu did not exceed the minimum detectable activity. Therefore, no quantitative values could be determined for 241Am, and the 239+240Pu activity concentration could be derived for only one sample: 0.5±0.1 Bq kg-1 (liver). The distribution pattern of these radionuclides in the tissues and organs of the roe deer could not be determined because of insufficient data. The HTO volumetric activity in the tissues and organs of the examined animals ranged from 2.6×10-2 to 77 kBq l-1; activity concentration of OBT, 3.0×10-2 to 16 kBq kg-1; and OBT-to-HTO ratios, 2.0×10-3 to 5.3×102. This ratio can serve as an indicator of how long the examined animals stay in radioactively contaminated ecosystems. Within the 'Degelen' site, the activity concentrations of 90Sr and tritium, in the form of HTO and OBT, are expected to be high in the bone tissues, soft tissues, and organs, respectively.

Assessment of environmental impacts from authorized discharges of tritiated water from the Fukushima site to coastal and offshore regions.

In August 2023, the long-planned discharging of radioactive wastewater from the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) started after the confirmation of its feasibility and safety. As this water contains elevated amounts of tritium even after being diluted, a lot of resources have been invested in the monitoring of the Fukushima coastal region where the discharge outlet is located. We compare the first 3H surface activity concentrations from these measurements (up to the end of November 2023) with the available background values to evaluate a possible impact of the long-term discharging on humans and environmental levels of the radionuclide of interest in the same or nearby area. From our results, we can conclude that the joint effect of horizontal and vertical mixing has been significant enough to reduce tritium concentrations at the monitored locations in the region close to the FDNPP port two days after the end of the respective phase of the discharging beyond the detection limit of the applied analytical methods (∼0.3 Bq L-1) which is by five orders of magnitude lower than safety limit for drinking water set by the World Health Organization (WHO). Moreover, the distant correlation analysis showed that tritium concentrations at stations located further than 1.4 km were very close to pre-discharge levels (∼0.4 Bq L-1). We also estimated that the 3H activity concentration in the offshore Fukushima region would be elevated by 0.01 Bq L-1 at maximum over a year of continuous discharging, which is in concordance with the already published modeling papers and much less than the impact of the FDNPP accident in 2011.

Development of Peptide-Based Probes for Molecular Imaging of the Postsynaptic Density in the Brain.

The postsynaptic density (PSD) comprises numerous scaffolding proteins, receptors, and signaling molecules that coordinate synaptic transmission in the brain. Postsynaptic density protein 95 (PSD-95) is a master scaffold protein within the PSD and one of its most abundant proteins and therefore constitutes a very attractive biomarker of PSD function and its pathological changes. Here, we exploit a high-affinity inhibitor of PSD-95, AVLX-144, as a template for developing probes for molecular imaging of the PSD. AVLX-144-based probes were labeled with the radioisotopes fluorine-18 and tritium, as well as a fluorescent tag. Tracer binding showed saturable, displaceable, and uneven distribution in rat brain slices, proving effective in quantitative autoradiography and cell imaging studies. Notably, we observed diminished tracer binding in human post-mortem Parkinson's disease (PD) brain slices, suggesting postsynaptic impairment in PD. We thus offer a suite of translational probes for visualizing and understanding PSD-related pathologies.

Tissue-free water tritium and non-exchangeable organically bound tritium concentrations in fish near coastline during and after operation of Japan's first nuclear fuel reprocessing facility.

The risks from radioactive wastewater release from nuclear facilities into the ocean are a global concern. Radioactive contaminants, such as tritium (3H), in both forms of tissue free water tritium (TFWT) and non-exchangeable organically bound tritium (NE-OBT), can be incorporated into marine biota and cause radiation doses to biota and future consumers. However, no studies have been conducted to measure both forms of 3H in marine fish as well as evaluate the residence time in the vicinity of a nuclear fuel reprocessing facility. Here, fish from a brackish lake and from the Pacific Ocean coastline of Japan, which are near such a facility, were collected between 2006 and 2021. The reprocessing facility was operational between 2006 and 2009, during which time about 300 times more tritiated water was discharged per year into the ocean compared to the period when the facility was not operational. During operation the annual release was 30 times higher than the treated water released annually from Fukushima Daiichi. As expected, TFWT and NE-OBT concentrations increased in marine fish during operations and had peak values of 3.59 ± 0.03 and 0.56 ± 0.03 Bq/L, respectively. Total dose rates to the fish were 36,000 times lower than the 10 µGy h-1 benchmark. Concentrations gradually decreased to pre-operational levels as the facility was turned off with NE-OBT taking twice as long. Fish sampled from the brackish lake tended to have more incorporated TFWT and NE-OBT concentrations than ocean fish. This indicates that ocean tides might have contributed to the accumulation of discharged tritiated water in the lake via a narrow water channel, which highlights the importance of examining all marine ecosystems in future operations. In both marine environments, the estimated committed effective dose using the highest observed data through ingestion was well below public limits (91,000 times lower).

Tritium-Labeled Nanodiamonds as an Instrument to Analyze Bioprosthetic Valve Coatings: A Case of Using a Nanodiamond Containing Coating on a Pork Aorta.

Coatings with xenogenic materials, made of detonation nanodiamonds, provide additional strength and increase elasticity. A functionally developed surface of nanodiamonds makes it possible to apply antibiotics. Previous experiments show the stability of such coatings; however, studies on stability in the bloodstream and calcification of the material in natural conditions have yet to be conducted. Tritium-labeled nanodiamonds (negative and positive) were obtained by the tritium activation method and used to develop coatings for a pork aorta to analyze their stability in a pig's bloodstream using a radiotracer technique. A chitosan layer was applied from a solution of carbonic acid under high-pressure conditions to prevent calcification. The obtained materials were used to prepare a porcine conduit, which was surgically stitched inside the pig's aorta for four months. The aorta samples, including nanodiamond-coated and control samples, were analyzed for nanodiamond content and calcium, using the radiotracer and ICP-AES methods. A histological analysis of the materials was also performed. The obtained coatings illustrate a high in vivo stability and low levels of calcification for all types of nanodiamonds. Even though we did not use additional antibiotics in this case, the development of infection was not observed for negatively charged nanodiamonds, opening up prospects for their use in developing coatings.

Optimization of a new liquid scintillation spectrometer for the measurement of environmental levels of 3H in water samples.

The activity concentration of 3H in water samples collected from places unaffected by nuclear activities or for human consumption can be very low. In these cases, determination procedures must achieve a Minimum Detectable Activity (MDA) low enough to ensure that 3H is accurately determined. In this paper, we present a method that uses a new Liquid Scintillation Spectrometer (LSC in what follows): the Quantulus GCT 6220. Furthermore, a new liquid scintillation cocktail, the ProSafe LT+, has been tested for 3H measurement, showing to be a good option for the determination of low levels of this radionuclide. The MDAs achieved are low enough to enable the measurement of very low levels of 3H in recent environmental water. The results obtained using a Quantulus GCT 6220 and Prosafe LT + are compared to those obtained with a Quantulus 1220 and Prosafe HC + as liquid scintillation cocktail.

Identification of tritium-contaminated areas using a tritium survey on the snow cover, the case of Semipalatinsk test site.

The article presents research findings on the content of tritium in the snow cover of test locations 'Balapan' and 'Sary-Uzen' in the territory of the Semipalatinsk Test Site. Based upon results, a number of tritium-contaminated areas were revealed. At the 'Balapan' site, elevated tritium concentrations in the snow were revealed in the vicinity of the 'Atomic lake' close to the 'Karazhyra' deposit and at the location of Lake 'Kishkensor'. At the 'Sary-Uzen' site, elevated tritium concentrations in the snow were discovered near borehole 101, at object 'Lazurite' and at an unnamed creek outflowing from the mountain range. Maximal of tritium activity concentration in the snow were 45 Bq/kg and 36 Bq/kg at the 'Balapan' and 'Sary-Uzen' sites, respectively. Background tritium concentrations in the snow cover of the STS territory were found to be 4.6-12 Bq/kg.

Interlaboratory comparison exercises for organically-bound tritium in the development of reference materials of environmental samples.

The complex behavior of tritium and the probability of increasing tritium concentrations released in the environment were the promotors for the research and development of laboratory methods that enable to accurately determine the various forms of tritium including organically-bound tritium (OBT) for public and regulatory assurance. The measurement of tritium is a key step for dose and risk assessment. The Cernavoda Nuclear Power Plant (NPP) in Romania improved preparation methods and tested environmental matrices for OBT analysis through intercomparison exercises. This paper describes the international Organically-Bound Tritium (OBT) intercomparison exercise, organized by the Cernavoda Nuclear Power Plant (NPP) in 2019-2020, using fruit sample (quince) from Cernavoda town. Evaluation of the results from the participating laboratories was performed using both robust analysis (Algorithm A) method described in the ISO 13528:2015 standard and ANOVA method. The results obtained are encouraging as an increased number of participating laboratories did not change the observed dispersion of the results for activity concentration level around 50 Bq/L of combustion water. The stability of the remaining sample will be checked in time to investigate its use as a reference material for OBT analysis at the environmental levels.

Estimation of accumulation potential for tritium in olive flounder on exposure of treated water derived from Fukushima Daiichi Nuclear Power Station: Tritium transfer from seawater and food chain into organically bound tritium in the targeted fish.

This study estimated the accumulation potential of tritium, a major radionuclide released from the Fukushima Daiichi Nuclear Power Station (FDNPS), into the olive flounder as organically bound tritium (OBT) using a computer simulation model. In this estimation, two transfer pathways into the OBT were assumed: formation from tritiated water (HTO) in the tricarboxylic acid (TCA) cycle, and ingestion of OBT through the food chain (from phytoplankton, small fish, to the flounder). The food chain structure was reconstructed based on fish growth model. The OBT concentration in the flounder was estimated on three scenarios: Tritium was supplied to the flounder as only HTO in seawater (Scenario 1), as HTO in seawater and OBT formed from HTO in the small fish (Scenario 2), and as HTO in seawater and OBT accumulated in the small fish through the formation and ingestion of OBT in phytoplankton (Scenario 3). The estimated OBT concentrations in the flounder were in the following order: Scenario 3 > 2 > 1. The ratio of the estimated concentration in Scenario 1 to that in Scenario 3 reached a certain value (66 % after a year from the start of HTO exposure), indicating that the tritium transfer from the seawater into the flounder more significantly contributed to this concentration than ingestions of the small fish and the phytoplankton. Additionally, the difference between the estimations of Scenarios 1 and 2 is significantly larger than that between Scenarios 2 and 3. This suggests that phytoplankton contributed weakly to the OBT concentration in the flounder compared to the small fish.

Characterization of tritiated JNJ-GluN2B-5 (3-[3H] 1-(azetidin-1-yl)-2-(6-(4-fluoro-3-methyl-phenyl)pyrrolo[3,2-b]pyridin-1-yl)ethanone), a high affinity GluN2B radioligand with selectivity over sigma receptors.

Here, we describe the characterization of a radioligand selective for GluN2B-containing NMDA receptors, 3-[3H] 1-(azetidin-1-yl)-2-(6-(4-fluoro-3-methyl-phenyl)pyrrolo[3,2-b]pyridin-1-yl)ethanone ([3H]-JNJ- GluN2B-5). In rat cortical membranes, the compound bound to a single site, and the following kinetic parameters were measured; association rate constant Kon = 0.0066 ± 0.0006 min-1 nM-1, dissociation rate constant Koff = 0.0210 ± 0.0001 min-1 indicating calculated KD = Koff/Kon = 3.3 ± 0.4 nM, (mean ± SEM, n = 3). The equilibrium dissociation constant determined from saturation binding experiments in rat cortex was KD of 2.6 ± 0.3 nM (mean ± SEM, n = 3). In contrast to the widely used GluN2B radioligand [3H]-Ro 25-6981, whose affinity Ki for sigma 1 and sigma 2 receptors are 2 and 189 nM, respectively, [3H]-JNJ-GluN2B-5 exhibits no measurable affinity for sigma 1 and sigma 2 receptors (Ki > 10 µM for both) providing distinct selectivity advantages. Anatomical distribution of [3H]-JNJ-GluN2B-5 binding sites in rat, mouse, dog, monkey, and human brain tissue was studied using in vitro autoradiography, which showed high specific binding in the hippocampus and cortex and negligible binding in the cerebellum. Enhanced selectivity for GluN2B-containing receptors translated to a good signal-to-noise ratio in both in vitro radioligand binding and in vitro autoradiography assays. In conclusion, [3H]-JNJ-GluN2B-5 is a high-affinity GluN2B radioligand with excellent signal-to-noise ratio and unprecedented selectivity.

Tritium distribution in the 'water-soil-air' system in the Semipalatinsk Test Site.

This article presents research findings on 3H in abiotic environmental compartments, specifically, the 'water-soil-air' system. All of the research areas are located within the Semipalatinsk Test Site (STS): the perimeter of the 'Degelen' site, the riverside zone of the Shagan river and the 'background' area-the southeastern part of the STS (SEP). As research progressed, numerical values of 3H and its species were revealed in various environmental compartments. The presence of 3H was registered not only in underground nuclear test locations but also in the 'background' area-SEP. Maximum 3H tritium concentrations in the water were detected at the 'Degelen' site (up to 57000±5000 Bq/kg) and the Shagan riv (up to 61500±6000 Bq/kg), in the air of the 'Degelen' site (up to 56±11 Bq/m3), in the soil of the 'Degelen' site (up to 5170±500 Bq/kg) and the Shagan riv (4100±400 Bq/kg) in the free water, at SEP (up to 1710±170 Bq/kg) in the organic constituent. Based upon all of the findings, 3H was found to be readily distributed in abiotic environmental compartments depending on certain conditions. Research suggests that water plays a key role in 3H migration processes in the natural system of interest. The second most but equally important constituent is soil and microorganisms of plant and animal origin living there. These assumptions are indirectly proven by research findings that show the HTO and HT air concentration dynamics depending on the sampling location.

Biodistribution of Drug/ADA Complexes: The Impact of Immune Complex Formation on Antibody Distribution.

The clinical use of therapeutic monoclonal antibodies (mAbs) for the treatment of cancer, inflammation, and other indications has been successfully established. A critical aspect of drug-antibody pharmacokinetics is immunogenicity, which triggers an immune response via an anti-drug antibody (ADA) and forms drug/ADA immune complexes (ICs). As a consequence, there may be a reduced efficacy upon neutralization by ADA or an accelerated drug clearance. It is therefore important to understand immunogenicity in biological therapies. A drug-like immunoglobulin G (IgG) was radiolabeled with tritium, and ICs were formed using polyclonal ADA, directed against the complementary-determining region of the drug-IgG, to investigate in vivo biodistribution in rodents. It was demonstrated that 65% of the radioactive IC dose was excreted within the first 24 h, compared with only 6% in the control group who received non-complexed 3H-drug. Autoradiographic imaging at the early time point indicated a deposition of immune complexes in the liver, lung, and spleen indicated by an increased radioactivity signal. A biodistribution study confirmed the results and revealed further insights regarding excretion and plasma profiles. It is assumed that the immune complexes are readily taken up by the reticuloendothelial system. The ICs are degraded proteolytically, and the released radioactively labeled amino acids are redistributed throughout the body. These are mainly renally excreted as indicated by urine measurements or incorporated into protein synthesis. These biodistribution studies using tritium-labeled immune complexes described in this article underline the importance of understanding the immunogenicity induced by therapeutic proteins and the resulting influence on biological behavior.

Characterization of geological and lithological features in the area proximal to tritium-contaminated groundwater at the Semipalatinsk test site.

The article presents the results of a study of groundwater contaminated with tritium in the vicinity of the 'Atomic Lake' - a crater filled with water as a result of a thermonuclear explosion on the territory of the former Semipalatinsk test site. This crater was created as part of an experimental thermonuclear explosion in 1965 with the aim of creating an artificial reservoir in arid areas. The study was carried out to identify the source of groundwater contamination near the crater formed from a thermonuclear test. There were two possible factors of pollution: the influence of contaminated water from the crater on the groundwater of the adjacent area, or groundwater polluting the water in the crater. It was necessary to find out the source of groundwater contamination and its connection with the water in the funnel. For this purpose, a study of the geological and lithological conditions of the territory adjacent to the funnel was carried out, which was carried out using drilling operations and hydrological measurements. Drilling work made it possible to study the depth of distribution of groundwater, hydrological work made it possible to determine the conditions of distribution of groundwater, as well as to take samples of groundwater. The assessment of the degree of groundwater contamination was carried out through water sampling and laboratory analysis. As a result, it was established that the geological and lithological conditions of the area limit the flow of contaminated groundwater to the water in the crater - the 'Atomic Lake'. Despite the fact that the waters in the crater from a thermonuclear explosion and the groundwater of the adjacent territory are contaminated with the radionuclide tritium, they have different sources of contamination and are not interconnected. Radionuclide analysis of groundwater showed that increased concentrations of tritium with a specific activity of up to 95 000 Bq/l are found in groundwater near the river bed. Shagan and this is due to the influence of the flow of groundwater coming from other parts of the landfill.

Development of methodology for identification and assessment of ecosystems with an underground source of tritium.

The article considers the issues of working out the suitable approaches for identifying zones with the presence of underground near-surface waters with increased concentrations of tritium discharged into a surface reservoir. The following methods were used as possible methods: determination of tritium content in snow cover, determination of tritium content in vegetation in the form of tritium of free water and organically bound tritium, determination of tritium content in river water and coastal vegetation. The studies were carried out at a previously identified site where groundwater with a tritium concentration of up to 6000 Bq/l is present, located in the vicinity of the city of Obninsk (Kaluga region, Russia). As a result of the conducted research, it was concluded that the analysis of the distribution of tritium in vegetation is an excellent methodological technique for identifying areas of location of near-surface underground waters contaminated with tritium. As a control parameter, both the concentration of tritium in the free water of plants and the content of organically bound tritium can be used. To detect underground tritium contamination the most promising use is the following indicator - the content of OBT in the shoots of woody plants. This parameter is very informative, and the sampling procedure for its determination has no seasonal restrictions, unlike such parameters as the content of tritium in grass and leaves, the content of tritium in snow cover, surface waters, which are preferably collected only in summer or winter. It should be noted that the control of surface waters of the groundwater discharge zone may not be a sufficiently informative indicator for identifying areas of polluted water inflow, since it depends on the ratio of the volumes of leaking polluted groundwater and the annual flow of the watercourse.

Hydrochemical systematics and isotope (δ18O, δD and 3H) variations of aquifer system of southern Bengal Basin: implications for groundwater pollution.

Hydrogeological, hydrochemical and isotopic traits of the groundwater in the Quaternary aquifer system in an urban-periurban locality within and encircling the Kolkata-Howrah twin city in the south Bengal Basin have been synthesised to explain the present- and paleo-hydrological processes, surface and groundwater interaction and mixing dynamics of contamination of groundwater. Rock-weathering, evaporation, ion-exchange and active mineral dissolution are the key processes commanding the groundwater chemistry. Freshwater flushing from the recharge zones had thinned the entrapped sea water which has generated the present-day brackish water by a non-uniform fusion. The best-fit line of the plots of δD and δ18O of groundwater samples displays a slope lower than that of local meteoric water line (LMWL) and global meteoric water line (GMWL) which hints that isotopic constitution of the groundwater of the present area is primarily formed by evaporation before or in the recharging process. A wide range of δ18O values in groundwater suggests that these waters are not blended enough to remove dissimilarities in isotope configuration of recharge water. This also suggests that many groundwaters are a result of mixing of present-day recharge and an older integrant recharged under previously cooler climatic conditions. The groundwater samples are more depleted of oxygen at the shallower level. The depleted samples cluster around the Tolly's nala (canal) where upper aquitard is missing or < 10-m thick. The tritium values range between 0.70 and 15.02 which indicate the occurrence of 'sub-modern', 'a mix of modern and sub-modern water' and 'modern water'. It indicates mingling of isotope-depleted water from the Hugli River by means of Tolly's canal with relatively less-depleted groundwater of Kolkata's late Pleistocene aquifer. The tritium values and Cl/Br ratio of groundwater samples adjoining Tolly's canal and elsewhere refer the direct infiltration of 'modern wastewater and freshwater' which mixes with the 'sub-modern water' in the aquifer system.