CC BY
4DOI 10.24412/CL-37135-2023-1-18-20
INVESTIGATION OF THE EFFECT OF PROLONGED EXPOSURE TO OCA-AEROSOL (GLYCERIN/PROPYLENE GLYCOL) ON RAT PLASMA BY RAMAN SPECTROSCOPY
ARINA SOKOVA1, EKATERINA LAZAREVA u, ALEXANDER POLOZHENKOV3, ARTEM
MYLNIKOV3
ALLA BUCHARSKAYA 1,2,3 AND VALERY TUCHIN 1,2,4
Scientific and Educational Institute of Optics and Biophotonics, Saratov State University, Russia Interdisciplinary Laboratory of Biophotonics, National Research Tomsk State University, Russia 3 Department of Pathological Anatomy, Saratov State Medical University, Russia 4 Laboratory of problems of laser diagnostics of technical and living systems Institute of Precision
Mechanics and Control RAS, Russia
ABSTRACT
Current problem of our time is the replacement of tobacco smoking with the use of devices with steam smoking mixtures. However, scientists have already proved that their use also has a negative effect on the human body causing irreversible changes [1-5]. Raman spectroscopy (RAMAN) is one of the methods that allows analyzing the molecular composition of a substance, in particular blood and plasma [6-7]. Various blood components have a significant effect on the Raman spectra [8-9]. The blood serum obtained after the separation of whole blood into fractions contains almost 90% water, about 6.6-8.5% proteins and other organic and mineral compounds, which are intermediate or final products of metabolism carried by blood.
The experiment was carried out using 14 female white laboratory rats of the Wistar line (weight 180 ± 30 g, aged 9 months). The animals were randomly selected and divided into 2 groups:1 - control group (without exposure); 2 - group with inhalation exposure to an OCA aerosol POD system. The content of glycerol and propylene glycol in the liquid intended for filling the POD system was 50/50%.
The impact of the POD system on rats was carried out using a specially designed installation, which is: a glass container with a volume of 0.003 m3 with two holes: the first hole for connecting the POD system, the second for the Janet syringe. The duration of one inhalation averaged 5-7 minutes, the total number of inhalations was 5 per day. The rest period of laboratory rats after one manipulation was 15 minutes. The animals were removed from the experiment on day 14. Excretion was carried out by decapitation, with the preliminary introduction of the dissociative injectable anesthetic Zoletil100. After that, blood sampling was performed.
The blood of laboratory rats was centrifuged for 15 minutes at 2000 rpm, after which blood plasma was taken and poured into separate test tubes. As a result, six blood plasma samples were obtained for the control group) and six samples for the group with inhalation exposure to OCA-aerosol (14 days).
To register the Raman spectra, hardware was used consisting of a spectrometer (Ocean Optics, QE65000), a diode laser (785 nm, laser energy 400 mw), a hand RAMAN probe (f = 7.5 mm) and a computer.
The RAMAN spectra after the analysis for the presence of peaks for the control group and the group of animals after exposure to OPA-aerosol are shown in Figure.
Figure: Raman spectrum for six samples of the control group (a) and the group exposed to inhalation
effects of OCA - aerosol (b)
The central position of the peaks was determined from the Raman spectra. Data for each of the groups are given in Table. To assess the impact of OCA, the impact efficiency coefficient was calculated EOC:
Eoc = /oca~Wo' x 100%,
I control
where Ioca - is the intensity of the selected line of the RAMAN spectrum for a group of animals after exposure to an OCA-aerosol, Icontroi - is the intensity of the selected line of the RAMAN spectrum for the control group. The results after averaging the data by groups are shown in Table and show by how many percent the intensity of RAMAN on the spectrum increased or decreased for the group exposed to inhalation effects of POD-systems aerosol relative to the intensity of the RAMAN spectrum for the control group. Since the resolution of the device is 10 cm-1, differences in the position of the peaks by a large amount may indicate possible changes in the molecular structure.
Table: Characteristics of RAMAN spectra for the control group and the group of animals exposed to inhalation effects of OPA -aerosol
Control group The group after the OPAaerosol EOC, % Component Raman shift, cm-1 Ref.
Raman shift, cm'1 Intensity, a.u. Raman shift, cm'1 Intensity, a.u.
422.85 6.30 422.85 4.94 -21.55 Histidine 422 [111
558.58 14.57 553.98 7.64 -47.55 Uracil 556 [111
620.18 15.36 622.44 10.21 -33.54 Glutathione 625 [111
642.74 13.08 651.73 9.24 -29.41 Tyrosine 643 [101
725.06 4.10 -- -- -- Acetyl coenzyme A 724 [111
837.75 11.08 839.88 10.39 -6.18 Tyrosine 830 [101
867.50 12.95 861.15 11.53 -10.94 D-(+)-Mannose 861 [111
-- -- 892.81 9.88 -- Palmitic acid 893 [111
911.67 13.21 913.76 11.65 -11.78 D-(+)-Trehalose 912 [111
982.02 4.70 -- -- -- Arginine 982 [111
1010.58 11.25 1010.58 9.43 -16.23 Phenylalanine 1003 [101
1046.96 4.83 -- -- -- Tryptophan 1046 [111
1063.00 4.14 -- -- -- Palmitic acid 1063 [111
1102.78 7.87 -- -- -- D-(+)-Trehalose 1102 [111
1173.22 7.58 1171.28 6.12 -19.23 Stearic acid 1173 [111
1272.36 10.73 -- -- -- Histidine 1271 [111
1342.91 13.29 1342.91 14.31 7.70 Hydrophobicity marker for tryptophan 1340 [101
1458.23 17.89 1458.23 18.74 4.72 D-Fructose-6-phosphate 1458 [111
1608.2 6.85 -- -- -- Phenylalanine 1609 [101
1626.62 10.25 1624.95 11.09 8.16 Amide 16301635 [101
1669.68 18.65 1668.04 18.15 -2.69 16601670 [101
According to the results obtained, it can be noted that the number of peaks in the intensity of the RAMAN spectrum in the group exposed to the OCA - aerosol significantly decreased in comparison with the control group. However, the group after the OCA - aerosol had one peak, which is missing from the reference group: this is the peak of intensity located at a RAMAN shift equal to 892 cm-1, which according to the literature data may correspond to the RAMAN band of palmitic acid [11].
The positions of the centers of most peaks have shifted slightly, but there are RAMAN bands that have not changed their position on the frequency scale. This phenomenon is observed at the RAMAN shifts equal to: 422.85, 1010.58, 1217.44, 1342.91, 1415.5 and 1458.23 cm-1.
A significant decrease in the intensity of RAMAN by 47.5% is observed in the band characteristic of uracil 556 cm-1, and in the experiment a change in the position of the center of this peak from 553 to 559 cm-1 is noted [11].
The reported study was funded by a grant under the Decree of the Government of the Russian Federation No. 220 of 09 April 2010 (Agreement No. 075-15-2021-615 of 04 June 2021)
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