See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/369856458

Implication of storage conditions on luminescence response from P.

phosphoreum in free and immobilized form

Article in Luminescence · April 2023

DOI: 10.1002/bio.4499

CITATIONS READS

0 14

4 authors, including:

Rajeev Ranjan Valentina Kratasyuk

Siberian Federal University Siberian Federal University
10 PUBLICATIONS 188 CITATIONS 154 PUBLICATIONS 1,426 CITATIONS

SEE PROFILE SEE PROFILE

All content following this page was uploaded by Valentina Kratasyuk on 11 March 2024.

The user has requested enhancement of the downloaded file.

Received: 23 December 2022 Revised: 27 March 2023 Accepted: 3 April 2023
DOI: 10.1002/bio.4499

RESEARCH ARTICLE

Implication of storage conditions on luminescence response

from Photobacterium phosphoreum in free and immobilized
form

Rajeev Ranjan 1,2 | Azhar Alamanova 1 | Antonina Sarangova 1 |

Valentina A. Kratasyuk 1,3

1
Laboratory of Bioluminescent
Biotechnologies, Department of Biophysics, Abstract
Institute of Fundamental Biology and
Bioluminescent bacteria in the form of a cell suspension for on-site hazard analysis
Biotechnology, Siberian Federal University,
79 Svobodny Prospect, Krasnoyarsk, Russia are not suitable as in vivo luminescence in free cells fluctuates and may lead to erro-
2
Department of Biomedical Science, Acharya neous results. Furthermore, the culture broth cannot be stored for long durations to
Narendra Dev College, University of Delhi,
Govindpuri, Kalkaji, New Delhi, India
continue sensing analytes as the luminescence ceases over time. Factors that affect
3
Institute of Biophysics SB RAS, Federal luminescence response include growth dynamism, and ambient environmental condi-
Research Center ‘Krasnoyarsk Science Center tions. The present study investigated the effect of storage conditions such as tem-
SB RAS’, Akademgorodok 50/50, Krasnoyarsk,
Russia perature (25 ± 2 C, room temperature; 4 C; and 20 C) and ambient aqueous
1
environment (M1: sucrose, 1.02 M; M2, bioluminescent media [tryptone, 10 g L ;
Correspondence
1 1 1 1
Rajeev Ranjan, Laboratory of Bioluminescent NaCl, 28.5 g L ; MgCl2.7H2O, 4.5 g L ; CaCl2, 0.5 g L ; KCl 0.5 g L ; yeast
Biotechnologies, Department of Biophysics, 1
extract, 1 g L ; H2O, 1 L]; M3, bioluminescent media and 95% glycerol, 1:1 ratio) on
Institute of Fundamental Biology and
Biotechnology, Siberian Federal University, the luminescence emission from the calcium alginate-immobilized Photobacterium
79 Svobodny Prospect, Krasnoyarsk 660 041,
phosphoreum (Sb) against the cells in free suspension for an extended period. The
Russia.
Email: rrandzhan@sfu-kras.ru; rajeev694@ results indicated that both the parameters that were undertaken markedly affected
gmail.com
the luminescence. In the study, Sb showed an enhanced luminescence emission than
Funding information the control up to 18.5-fold and for a prolonged period which can be efficiently
Gastronomic Institute of Siberian Federal
utilized for rapid biosensing of hazardous materials.
University; Gastronomic R&D Park for the
Development Programme of the Federal State
Educational Institution of Higher Education KEYWORDS
‘Siberian Federal University’, Grant/Award bioluminescent bacteria, immobilization, luminescence response, storage conditions
Number: (2021–2030)

1 | I N T RO DU CT I O N of the FMNH2 that forms an intermediate called 4α-hydroperoxyflavin

The use of bioluminescent bacteria (BB) and their enzyme system in
screening hazardous materials is widely accepted as they offer rapid
results without the requirement for expensive instrumentation,
dedicated laboratory space, and skilled personnel [1, 2]. BB emit
luminescence (490 nm) via an intracellular biochemical reaction. The
reaction components include bacterial luciferase (BLuc, 79 kDa, a
heterodimer with subunits designated as α, and β), a long-chain
aliphatic aldehyde having a carbon chain length in the range of
C10–C16, reduced flavin mononucleotide (FMNH2), and molecular
oxygen (O2) [3, 4]. The long-chain aldehyde binds to the 4α position
(FMN-4a-hydroperoxide) that releases energy in the form of light [5].
The luminescence emission is continuous in BB as they regenerate
the oxidized products such as the corresponding long-chain acid via
fatty acid reductase complex, while the FMN regains its reducing
power via an NADH-mediated oxidoreductase (NADH: FMN
reductase) [6].
In vivo, the luminescence of BB is sensitive toward analytes that
are generally hazardous [1]. The bioluminescence inhibition (BLI) is
observed within minutes and is dose-dependent. The reasons for BLI
may be attributed to (a) disruption of vital transport channels embed-
ded in the cell wall, (b) binding to enzyme residues such as –SH, or

Luminescence. 2023;1–5. wileyonlinelibrary.com/journal/bio © 2023 John Wiley & Sons Ltd. 1


2 RANJAN ET AL.
(c) the induction of reactive oxygen species [7, 8]. However, there are
limitations to the utility of these luminescent microorganisms. The
immediate concern is the marked variability in luminescence emission
during experimental analysis [9]. The dynamic nature of the bacteria
in suspension and thereof sensitivity toward molecular oxygen [10],
temperature, and the ambient environment are the presumptions for
such behaviors [11]. Another difficulty associated with the application
of free BB is their viability over an extended duration. Previously, the
use of freeze-dried preparations of BB was adopted for hazard assess-
ment. The process of freeze drying is rather difficult, time intensive,
and requires careful optimization of different parameters including
co-additives such as trehalose, sorbitol, monosodium glutamate,
glucose, and Tween 80. Although the reactivation of freeze-dried BB
requires a short incubation time of 15 to 30 min, during incubation,
the co-additives, humidity, and exposure to air were observed to neg-
atively affect the luminescence response [12–14].
Therefore, freeze-dried BB preparations could not be utilized for
real-time hazard assessment.
To circumvent such limitations, whole-cell immobilization of BB
was attempted to slow down the metabolic activities of viable cells
without affecting luminescence emission. The use of agar and thick
silicate films for the immobilization of recombinant BB was described
[15]. However, the immobilization of wild-type BB is rather difficult.
Osmolarity considerations must be accounted for to avoid osmotic
shock. Comparative assessment for luminescence response in agar-
, calcium alginate-, and carrageenan-immobilized wild-type Photobac-
terium leiognathi cells has also been reported previously and was
utilized for hazard assessment [16]. However, reports on the role of
the ambient media surrounding the BB have not been well studied.
In the present research, three different ambient environments
were selected such as 1.02 M aqueous sucrose solution (M1), biolumi-
nescent media (M2), and a 1:1 ratio of bioluminescent media and 95%
glycerol (M3) for studying the effect of calcium alginate-immobilized
wild-type Photobacterium phosphoreum (P. phosphoreum) on the lumi-
nescence response in comparison with cells in suspension form.
2 | MATERIALS AND METHODS
Tryptone, yeast extract, sodium alginate, sodium chloride (NaCl), mag-
nesium chloride (MgCl2.7H2O), sucrose, anhydrous calcium chloride
(CaCl2), and potassium chloride (KCl) were purchased from Medigen,
Novosibirsk, Russia. The cotton mesh was locally purchased from a
medical store. The wild-type 1889 strain of P. phosphoreum was
obtained from the culture collection facility at the Institute of Bio-
physics (IBP), Siberian Branch (SB), the Russian Academy of Sciences
(RAS), Krasnoyarsk Science Center (KSC) SB RAS.
A loopful of inoculum was taken from a freshly prepared slant of
P. phosphoreum and put into a 250-ml Erlenmeyer flask containing
50 ml of sterile broth (bioluminescent media: tryptone, 10 g L 1
; NaCl,
1 1 1 1
28.5 g L ; MgCl2.7H2O, 4.5 g L ; CaCl2, 0.5 g L ; KCl 0.5 g L
1
yeast extract, 1 g L ; H2O, 1 L) inside a class II biohazard safety
cabinet (ESCO, Singapore). The flask was incubated for a period of
15227243, 0, Downloaded from https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4499 by RAJEEV RANJAN - Siberian Federal University , Wiley Online Library on [27/04/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
18 h at 25 C under mild shaking of 75 rpm using an incubator shaker
(WiseCube WIS-20R, Witeg, Germany). Post-incubation, 1 ml of the
cultivated broth was transferred in three microfuge vials and sub-
jected to centrifugation at 4500 g at 20 C using a benchtop centrifuge
(Eppendorf 5430 R, Hamburg, Germany). The supernatants were care-
fully discarded while the pellets were reconstituted with the ambient
media designated as M1, M2, and M3. The optical density at 660 nm
was determined using a UV–Vis spectrophotometer (Cary 60, Agilent
Technologies, USA) for the reconstituted samples individually using a
quartz cuvette of 1 ml volume (Hellma Analytics, Müllheim, Germany).
Before the OD measurements, the blank readings were taken for the
three ambient solutions (M1, M2, and M3) and the baselines were
normalized.
2.1 | Whole-cell immobilization
In total, 0.1 g sodium alginate was weighed using a precision analytical
balance (Ohaus, Parsippany, USA) and subjected to a 20 ml volume
glass beaker containing 5 ml of distilled water. A PTFE magnetic stir
bar was immersed in the beaker kept on an IKA RH basic 2 magnetic
stirrer (Staufen, Germany) under mild shaking until the complete dis-
solution of sodium alginate in water to form a viscous 2% solution of
sodium alginate. Care was taken to avoid the formation of trapped air
bubbles inside the gel. Furthermore, 1 ml of 2% sodium alginate solu-
tion was aspirated using a broad end tip connected to a micropipette
and mixed homogenously with the 1 ml culture suspensions such as
M1, M2, and M3 individually. After thorough mixing, the viscous cell
suspension was allowed to pass through the tip-end orifice into a
chilled aqueous solution of 0.34 M calcium chloride. Immediately, the
viscous drop-hardened spherical beads were separated from the pre-
chilled calcium chloride solution using a cotton mesh, and immersed
into the respective ambient solutions (M1, M2, and M3).
2.2 | Luminescence measurements
The cells in suspension (control) and the immobilized form under dif-
ferent ambient environments (M1, M2, and M3) and temperatures
(25, 4, and 20 C) were recorded for their luminescence response
(Relative luminescence unit, RLU) in triplicate (n = 3) for 7 days using
a benchtop luminometer (Glomax 20/20, Promega, USA) and the
results were expressed as mean ± standard deviation (SD). Before the
luminescence measurements, aliquots of the samples kept at 4 and
20 C were allowed to attain 25 C.
In brief, 10 μl of sample from the shortly vortexed vials (M1, M2,
and M3) was transferred to a borosilicate luminometric cuvette pre-
filled with 240 μl of the respective ambient media. It was estimated
that 20 μl of the viscous cell suspension formed one immobilized
spherical structure (Spherical bead, Sb) and was equivalent to 10 μl of
; the cells in free form. Therefore, luminescence measurements in
immobilized cells were performed by immersing a single Sb in lumino-
metric cuvettes individually containing 230 μl of the ambient media

RANJAN ET AL.
(n = 3). The luminometric cuvettes containing the samples were
vortexed for a period of 5 s and inserted into the sample holder imme-
diately to perform the luminescence measurements.
3 | RESULTS AND DISCUSSION
3.1 | Optical density measurements
The optical density at 660 nm (OD660) for the cells in the vials
designated as M1, M2, and M3 were estimated to be 0.906, 0.909,
and 0.907 respectively. The OD660 measurements for the cells in
three different ambient media suggested a rough estimate of
P. phosphoreum cells in these vials and were presumed to be uniformly
distributed [17].
3.2 | Bioluminescence response of P. phosphoreum
in ambient media
Luminescence measurements in the vials M1, M2, and M3 before
immobilization showed the RLU values of 9.12  107 ± 1.2  105,
1.03  108 ± 1.43  107, and 1.95  104 ± 1.04  103 respectively
designated as 0 h (n = 3). The luminescence response of
P. phosphoreum cells was affected by the ambient environment and
can be arranged as M2 > M1 > M3. M3 was found to largely affect
the luminescence of these cells. The possible reason may be differ-
ences in osmolarity (iC, where i = the van ’t Hoff factor, and C = the
molarity of the solution) of the M3 and the intracellular environment
of P. phosphoreum cells [18]. In total, 1.02 M aqueous sucrose solution
(M1) was selected in the study as this led to peak luminescence
emission by the P. phosphoreum cells when different molar concentra-
tions of sucrose solution were used as the ambient media. A rough
estimate for the osmolarity of M2 solution can be calculated as
iNaClCNaCl + iMgCl2.7H2OCMgCl2.7H2O + iCaCl2CCaCl2 + iKClCKCl
(2  0.487 + 3  0.022 + 3  0.004 + 2  0.007) that sums up as
1.06 M. The results highlight the role of osmolarity in affecting lumi-
nescence levels. In M3, the osmolar strength of the solution can be
calculated as 1/2 (1.06 + iGlycerol C Glycerol) shows 5.68 M. The reason
for the selection of 50% glycerol (95% w/V) in the experimental
analysis was to understand how the P. phosphoreum responded to the
hyperosmotic environment of 5.68 M and the role of glycerol as a

cryoprotectant for P. phosphoreum at 20 C that might compensate
for luminescence loss.
3.2.1 | Bioluminescence response in M1
Luminescence levels in Sb for M1 were calculated to be
6.7  105 ± 7.5  104 (n = 3) at Day 1 (24 h post-storage) while the
luminescence in control was 2.3  10 ± 6.0  10 , merely 3.4% of
4 3
the Sb. The Day 2 to Day 6 measurements in control samples showed
a bell-shaped curve and were below 50% of luminescence in
15227243, 0, Downloaded from https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4499 by RAJEEV RANJAN - Siberian Federal University , Wiley Online Library on [27/04/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
3
comparison with Sb in all cases (Figure 1a). The study indicates the
role of immobilization in extending the luminescence profile of
P. phosphoreum cells and can be effectively utilized for hazard assess-
ment [19]. The initial rise of luminescence in the control at Day 3 may
be due to a rise in the number of cells, however, subsequent ceasing
luminescence might reflect a deficit in essential nutrient requirements.
At 4 C, the Sb showed an RLU response of 5.1  106 ± 1.8  105
while the control sample had comparatively higher luminescence
(5.3  106 ± 5.1  105) on Day 1. The luminescence response at Day
2 to Day 6 period for the cell suspension at 4 C was significantly
lower than Sb (Figure 1b). When Figure 1a is compared with
Figure 1b, the role of storage temperature of 4 in modulating lumi-
nescence from Sb can be demonstrated, while cells in free suspension
failed to emit enough luminescence to perform relevant bioassays.
A similar trend was observed when the storage temperature was
20 C (Figure 1c). Although the luminescence levels decayed expo-
nentially in Sb, it could provide scope for bioassays until a decent
period of 6 days (Table S1).
3.2.2 | Bioluminescence response in M2
The mean RLU value for Sb was 5.2  106 ± 6.2  105 after 24 h at
25 C and the control samples had 1.7  107 ± 1.4  106 after 24 h
(Day 1) (Table S2). The Day 2 to Day 6 RLU data showed a significant
decline in luminescence response in cell suspension (control) as well
as Sb. However, the RLU response in Sb was 18.5 times higher than
the control on Day 2 and more than three times higher in subsequent
measurements (Figure 2a).
The Day 1 RLU measurements at 4 C showed the biolumines-
cence response as 2.9  107 ± 2.2  106 for Sb and
6.8  10 ± 1.0  10 in the case of cells in suspension form (control).
7 6
Day 2 also observed elevated luminescence in the control than Sb. In
contrast, Day 3 RLU data suggested 1.82 times higher luminescence
in Sb than control and subsequent rise on Day 5 and Day 6 by 3.64
and 2.74 times respectively (Figure 2b).
At 20 C storage conditions, bioluminescence response from Sb
was estimated to be higher than control from Day 2 onwards up to
11.61 times. However, the Day 1 RLU data showed the RLU values of
Sb as 2.8  107 ± 2.6  106 and 7.8  107 ± 1.24  106 in control
(Figure 2c).
The data in this section suggested that the luminescence levels in
M2 at 4 and 20 C could not sustain luminescence to levels that
might be utilized for biosensing purposes. The Sb performed better at
25 C in M2. Moreover, cells in free suspension showed an elevated
response of luminescence than Sb at a few days' measurements. The
possible reason may be the destabilization of calcium-immobilized
P. phosphoreum cells (Sb) at lower temperatures in M2 as the ambient
environment has many ionic species that might interfere with the
structural integrity of the Sb [20]. The justification for the marked loss
of luminescence in M2 vials kept at 25 C is due to the enhanced met-
abolic activity of P. phosphoreum that could result in the accumulation
of waste by-products.
 15227243, 0, Downloaded from https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4499 by RAJEEV RANJAN - Siberian Federal University , Wiley Online Library on [27/04/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
4 RANJAN ET AL.

F I G U R E 1 Bioluminescence response of calcium alginate-immobilized Photobacterium phosphoreum cells (spherical bead, Sb) and
P. phosphoreum cell suspension (control) in 1.02 M aqueous sucrose solution (M1) up to a period of 6 days at (a) 25 C, (b) 4 C, and (c) 20 C.

F I G U R E 2 Bioluminescence response of calcium alginate-immobilized Photobacterium phosphoreum cells (spherical bead, Sb) and
P. phosphoreum cell suspension (control) in bioluminescent media (M2) up to a period of 6 days at (a) 25 C, (b) 4 C, and (c) 20 C.

F I G U R E 3 Bioluminescence response of calcium alginate-immobilized Photobacterium phosphoreum cells (spherical bead, Sb) and
P. phosphoreum cell suspension (control) in 1:1 ratio of bioluminescent media and 95% glycerol (M3) up to a period of 6 days at (a) 25 C, (b) 4 C,
and (c) 20 C.

3.2.3 | Bioluminescence response in M3
The Sb and the cell suspension (control) in M3 showed an entirely dif-
ferent trend in comparison with M1 and M2. Firstly, the luminescence
response at all the temperatures studied was much lower than their
counterparts.
At 25 C, the Sb emitted a mean RLU of 3649.3 ± 294.3 and was
1.37 times higher than the control post 24 h (Day 1) (Table S3). From
Day 4 to Day 6, luminescence levels in Sb increased approximately
four times compared with the control (Figure 3a). A similar observa-
tion was seen when the Sb and the control samples were stored at
4 C (Figure 3b).
In terms of luminescence emission, 20 C provided a suitable
environment for Sb than control. After 24 h (Day 1), the mean RLU
value for Sb was 10,044.7 ± 1635.0, 2.76 times higher than the con-
trol (Figure 3c). More than 1.6-fold luminescence emission continued
for Sb than control during subsequent measurements. The possible
reason for comparatively lower luminescence levels in M3 than M1
and M2 for Sb and control has been discussed in the previous section.
However, the point of consideration is their prolonged and relatively
stable luminescence from Day 1 until the end of the experimental
analysis. It can be argued that even at 5 M solutions, a portion of cells
can survive (assuming that lowering of RLU indicates loss of cell
viability) and emit luminescence signals that can be applied for
biosensing.
4 | CONC LU SION
The present study emphasizes the role of ambient media in affecting
the luminescence response from BB in free suspension as well as
immobilized form (Sb). Sb showed upliftment in the luminescence
response than control and can be efficiently utilized for hazard assess-
ment over a considerable period (up to 6 days in the present study)
when kept at 25 C. Therefore, Sb has the potential to be utilized for
real-time pre-screening of relevant analytes without the requirement

RANJAN ET AL.
of additional laboratory reagents and equipment. Sb is conveniently
prepared within a short period without the use of complicated proce-
dures and does not require pre-incubation, unlike freeze-dried BB.
AUTHOR CONTRIBUTIONS
Rajeev Ranjan: Conceptualization; investigation; data curation; roles/
writing—original draft; writing—review and editing. Azhar Alamanova:
Investigation; methodology. Antonina Sarangova: Formal analysis;
investigation; methodology. Valentina A. Kratasyuk: Supervision;
validation; project administration; formal analysis; writing—review and
editing.
ACKNOWLEDGMENTS
The authors acknowledge the Gastronomic Institute of Siberian
Federal University and the project Gastronomic R&D Park for the
Development Programme of the Federal State Educational Institution
of Higher Education ‘Siberian Federal University’ (2021–2030).
CONF LICT OF IN TE RE ST ST AT E MENT
The authors declare no competing conflicts of interest.
DATA AVAI LAB ILITY S TATEMENT
The data that supports the findings of this study are available in the
supplementary material of this article.
ORCID
Rajeev Ranjan https://orcid.org/0000-0003-4692-9504
Azhar Alamanova https://orcid.org/0000-0001-7493-1199
RE FE R ENC E S
[1] E. N. Esimbekova, V. P. Kalyabina, K. V. Kopylova, I. G. Torgashina,
V. A. Kratasyuk, Talanta 2021, 233, 122509.
[2] E. M. Kolosova, O. S. Sutormin, L. V. Stepanova, A. A. Shpedt, N. V.
Rimatskaya, I. E. Sukovataya, V. A. Kratasyuk, Environ. Technol. Innov.
2021, 24, 101842.
[3] S. E. Abdel Aal, A. M. Dessouki, H. H. Sokker, J. Radioanal. Nucl. Chem.
2001, 250, 329.
View publication stats
15227243, 0, Downloaded from https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4499 by RAJEEV RANJAN - Siberian Federal University , Wiley Online Library on [27/04/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
5
[4] S. Girotti, E. N. Ferri, M. G. Fumo, E. Maiolini, Anal. Chim. Acta 2008,
608, 2.
[5] E. Brodl, A. Winkler, P. Macheroux, Comput. Struct. Biotechnol. J.
2018, 16, 551.
[6] E. Esimbekova, V. Kratasyuk, O. Shimomura, Bioluminescence:
Fundamentals and Applications in Biotechnology, Vol. 1, Springer Berlin,
Heidelberg 2014, 67.
[7] E. T. Hwang, J. H. Lee, Y. J. Chae, Y. S. Kim, B. C. Kim, B. I. Sang,
M. B. Gu, Small 2008, 4, 746.
[8] M. Woutersen, S. Belkin, B. Brouwer, A. P. van Wezel, M. B. Heringa,
Anal. Bioanal. Chem. 2011, 400, 915.
[9] L. Camanzi, L. Bolelli, E. Maiolini, S. Girotti, D. Matteuzzi, Environ. Tox-
icol. Chem. 2011, 30, 801.
[10] Y. Li, S. Wang, X. He, S. Li, T. Zheng, Y. P. Chen, H. Cui, W. Wang,
Chem. Sci. 2021, 12, 12400.
[11] G. C. Gil, R. J. Mitchell, S. T. Changand, M. B. Gu, Biosens. Bioelectron.
2000, 15, 23.
[12] M. B. Gu, S. H. Choi, S. W. Kim, J. Biotechnol. 2001, 88, 95.
[13] S. H. Choi, M. B. Gu, Biosens. Bioelectron. 2002, 17, 433.
[14] J. Yang, S. Hu, A. Liao, Y. Weng, S. Liang, Y. Lin, Food Sci. Nutr. 2022,
10, 1841.
[15] R. J. Mitchell, M. B. Gu, Biosens. Bioelectron. 2006, 22, 192.
[16] R. Ranjan, N. K. Rastogi, M. S. Thakur, J. Hazard. Mater. 2012,
225, 114.
[17] E. A. Meighen, Luminescence 1999, 14, 3.
[18] V. L. Jennings, M. H. Rayner-Brandes, D. J. Bird, Water Res. 2001, 35,
3448.
[19] M. M. Calabretta, A. Lopreside, L. Montali, M. Zangheri, L. Evangelisti,
M. D'Elia, E. Michelini, Anal.Chim. Acta 2022, 1200, 339583.
[20] T. Mehrotra, S. Dev, A. Banerjee, A. Chatterjee, R. Singh, S. Aggarwal,
J. Environ. Chem. Eng. 2021, 9, 105920.
SUPPORTING INF ORMATION
Additional supporting information can be found online in the Support-
ing Information section at the end of this article.
How to cite this article: R. Ranjan, A. Alamanova,
A. Sarangova, V. A. Kratasyuk, Luminescence 2023, 1. https://
doi.org/10.1002/bio.4499