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Detection of Bisphenol in Pregnant Women and its Role in Abortion

Jaleel Ibrahim Asaad1, Habeeb Waseel Kadhum Shubber2,Nahidh Mohammad jabur Almamori3,Ayad M.J. Al-mamoori4 ,Rasha Nayyef Ali Assi Banwawi 4

1Biotechnology Research Center, AL-Nahrin University, Iraq.

2University of Al-Qadisiyah/College of Science/Biology Department 3 Babil health Directorate, Ministry of Health, Iraq

4Department of Biology, College of Science, University of Babylon, Babylon, Iraq Corresponding author: [email protected]

ABSTRACT

Bisphenols as synthetic chemicals, been utilized for detection in environmental and human samples. In case of the sensitivity analyses, the bisphenols are detected using a chromatogram along with time and amount of bisphenol. In this paper various study is performed to detect the bisphenol in pregnant women and the effects of bisphenol role in abortion i.e. loss of pregnancy. The bisphenols have a adverse effect in the environment and their presence could lead a negative impact in the future born babies. The various methods of the bisphenol detection is discussed in this paper along with the flow rate and the chemical composition CH3CN , D.W and phosphate buffer in the ratio of 66, 33 and 7 and the detector is considered as FLO (ex = 360nmn and em = 460nmn). The sample test is conducted within the living organism and outside the living organisms.

Keywords: Bisphenol, sensitive analysis, pregnant women, estrogen receptor, bisphenol detector.

1. INTRODUCTION:

A chemical component termed as bisphenol is the major component that is extensively used in various consumer products such as the use of polycarbonate plastics and epoxy resins are highly exploited.

These use of polycarbonate plastics and epoxy resins are highly treacherous to environment and organisms. The aggregations are stated in the blood specimen of the humans from the picomolar nanomolar levels within the water also in urine specimens and at picomolar and nanomolar levels in water and even reached at higher levels in human blood samples and urine samples as the usage of these bisphenols in day to day life is improvised in a drastic manner.(1).

Bisphenol is considered as endocrine disruptors and it may alter the collection of the various corporeal functions of the human beings that are being associated (prone) with the health disorders like obesity, diabetes, reproductive problems, heart related diseases, child birth imperfections, breast malignant growth and other various diseases(2; 3). Due it’s adverse effect the chemical bisphenol is outlawed in various products. The alternatives of bisphenol counterparts like bisphenol S (BPS) and bisphenol F (BPF) are being extensively used in place of the bisphenol in the consumer products where the use of polycarbonate plastics and epoxy resins. The samples of BPS and BPF are detected in human samples and compared with bisphenol detections and concluded that the BPS and BPF are less harmful to humans depending on the usage factor on the different conditions. (4; 5). The samples of the bisphenols in the human beings are in higher concentrations than expected in most of the Asian countries. However the bisphenol alternatives in large proportions are utilized so far as the countries like Japan, China and India are notified to the concentrations of nearly 11.4 nM and the these type of toxicological information is limited for the provided anti - estrogenic and anti – androgenic activities of the bisphenol and it’s alternatives BPS and BPF can intervene with the thyroid hormone and thyroid stimulating hormone which is also crucial for the pregnant women for the children and to protect them from abortion is being affected by the bisphenol (6). The over usage of the bisphenol is hazardous that is being regulating the THs hormones that are highly crucial for

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the usual improvement, progress and metabolism vertebrates. The receptors that resides on thyroid hormonesare exert the actions predominantly by binding on the thyroid response elements in the genes of the thyroid followed with the dynamic recruitment or shedding the thyroid complex hormones(7). Noxious effect of bisphenol is determined by the analytical methodologies based on colorimetric, spectroscopic, chemiluminescence, and other methods are being implemented for the analysis of the bisphenol composition. However, the above mentioned are being advanced to assist effective way for the judicious and sensitive analysis of bisphenol. As these have few cons as the purchasing cost of the instrument is high expenditure, time consuming . the sample detection also should be assisted under a specialist for the operation of the systems. Electrochemical methods are also available with the extraordinary features that can be stated as the alternative way for the bisphenol detection. The electrochemical way such as the use of electrodes are being intensified and the various study are being carried out as the bisphenol exhibits the destitute electrochemical response due to sluggish electrode kinetics and formation of electro – polymerized film on the electrode leading to the electrode fouling, low sensitivity and reproducibility(8).

Then, the thyroid dependent GH3 cell proliferation assay that is living outside the organism (vitro), that is beingintermittently utilized to examine the thyroid disruptors within the cellular stage. Therefore, the gene thyroid response with pelophylax nigromaculatus transcript assay that is being used in the previous TH signalling disrupting activities of the bisphenols inside the living organism (vivo), particularly at the environmentally relevant concentrations. TH signalling disarray activities of the BPS and BPS are being compared with bisphenols as the formation way provides improvised and maximum quality of nanocomposites within the enhanced polymer – Nano tube synergy. Thus the present approach of the simplistic electrochemical preparation of poly – CTAB / MWCNTs nanocomposite on PGE application for the electrochemical analysis of bisphenol and till date the electrochemical approach of bisphenol is not implemented. The polymer based film of the cationic surfactant combined witgh mWCNTs on the surface showed distinct structure and allowed good electro – catalytic activity and high sensitivity towards the oxidation of the bisphenol (BPA). The surface characterization of the PGEs is carried out by the scanning of the electron microscope (SEM) and the electrochemical impedance spectroscopy (EIS) to prove the formation of the composite on to the surface. Moreover, the developed material detection sensitivity analysis has been applied for the perseverance of bisphenol in pregnant women and the bisphenol effect in the abortion in pregnant women with samplesis being carried out to real matrices(9).

2. Detection methods and materials 2.1 With chemicals

The bisphenol and Triiodothyronine (T3) with purity of 95% were attained from across organics. With the NaOH the T3 can be mixed within to acquire a approximate 6.60 g / L stock solution from the water (ultrapure).A stock solution of bisphenol is processed by dissolving in DMSO (Dimethyl sulfoxide). BPF, purity of more than 99% and BPS purity of 98% acquired within chemicals that are assorted.

Extra pure silver nitrate can be obtained from Merck, Trisodium citrate (which is 98% anhydrous), 4 – ethylresorcinol (which is 98%), 4 – tert – butylphenol (which is 97 %), 4 – ethylresorcinol (which is 98%), 4 – tert – butylca – techol (which is 99% anhydrous) and with purity of more than 99% sodium nitrate endured from Acros Organics. Pyridine (which is 99% anhydrous), bisphenol B (4, 4’ – (butane – 2, 2 – diyl) di – phenol) and bisphenol F (bis (4 – hydroxyphenyl) methane which is 98%) are obtained from Sigma Aldrich. 4, 4’ – iso – propylidenediphenol, more than 99% (bisphenol A) is generated from the assorted chemicals. Water (ultrapure) is engendered within the Milli – Q structure for the formation of the trisodium citrate, pyridine, silver nitrate and sodium nitrate mixtures. The grade of LC / MS water from Bio – solve is generally utilized for the formation within the mixtures of the sensitivity analysis. The mixtures ofof 4 – ethylresorcinol 4- tert – butylphenol and 4 – tert – butylcatechol were blended with tap water at the 10 mg / L concentrations in such a way that to execute preparatory test. The stock blends of the bisphenols are processed and diluted at distinctive concentrations (0.25 mg / L to 100 mg / L) under a tap water of 10 mg / L concentration in a Branson 2510 ultrasonic bath with 40KHz and a power potential of 80W for about 120 minutes. The similar experiment can be enforced to real samples, that which water

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(tap water about 500ml) is implanted in the ultrasonic bath for 120 minutes after weighing 500mg of each sample.

2.3 Molecular docking

AutoDock is being utilized to replicate the interaction of bisphenol effects, the BPF and BPS along TR – LBDs are conserved rigorously, as long as every torsional bonds appertaining to every tissues abide to be unoccupied. With the help of the Lamarckian genetic algorithm docking is accomplished. in case of perplexing, ten individual docking tissues are handled, the composure approach along with minimal energy was preferred.

2.4 Coactivator recruitment assay

During the pre- experiment it is observed that the bisphenols appear to display the agonistic effects on the TH signalling in several assays. Therefore, an assessment of the potential of BPS and BPF to recruit the coactivator assay, with the following method is being employed in the study. Then,fluroesence polarization measurement was conducted to detect the binding of the peptide probeto TR –LBDs. T3 (1mM) was tested as the positive control. The assay was conducted in triplicate for each treatment.

2.5 Other methods

In the process of detecting the bisphenol a 300 µL mixture of phosphate buffer was induced in the centrifugal machine tube., with an inclusion about 50 µL of DNA – nanoparticle complexes and the 50 µL of AHN – labelled aptamer. During full mix, the blend mixture is allowed to incubate for about 20 minutes at room temperature ( 25 to 28 degree Celsius). then the centrifuge tube is removed within the incubator and with help of magnet a magnet separation is made. At about, theconcentration range of 320µL of phosphate buffer is supplemented to the centrifugal machine tube to re-append the DNA – Nano particle in the mixture solution and then about 80 µL of 100 ng / mL bisphenol is combined to the blend and the mixture solution is permitted to incubate at room temperature for about 10 minutes. During the end of the method, the centrifugal machine tube is allowed on the magnet to disconnect the DNA – Nano particle within the assorted blend mixture. Lastly, the supernatant fluorescence intensity is achieved with the help of magnetic separation was noted and the bisphenol is detected and measured by HPLC(Shimadzu type) in Biology Dept.college of Science.

3. Results

The sample bisphenol is tested under various statstical methods with the help of a chromotogram at a 1 ml / min of flow rate. Figure 1 is illustrated wth the information related to bisphenol (5 ppb).

Figure 1: Bisphenol 5ppb.

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Figure 2, describes about the sample information 1, where the flow is same ad the ret. time is varied from 5.356 to 4.759 is described in chromatogram below.

Figure 2: Variation in peak time.

Now, we have the changes in the area per cent to 67.775 % and the changes are described here in the chromatogram below.

Figure 3: Area change with bisphenol.

Similarly, the changes in the ret. time and the peak area per cent is shown in the chromatogram are in the figure 4, figure 5, figure 6, figure 7, figure 8, figure 9 and figure 10 respectively.

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Figure 4: Variation in time and area

Figure 5: Sample 5 with variation in area.

From the samples, it is clear that, for the different experimental analysis the changes in bisphenol is occurring with the flow rate of 1 ml / min. The following table resembles the experimental data that is executed among the control ppb (parts per billion) and the patients ppb (parts per billion) is stated here.

4. Conclusion and discussion

In this paper, the bisphenol and the effects are adverse in the pregnant women as they lead to abortion in them and the detection of bisphenol with various materials and there is discussed(10).

Bisphenol is a endocrine - discombobulate chemical that endangers by disturbing the normal hormone excretion owing to the interference of human endocrine system. The correspondent of estrogen which is in a particular structure and action, the bisphenol reacts with the estrogen receptor within the food chain(11), that tends to demolish normal action of the estrogen, like production, release, translocation and metabolism in organisms(12). Therefore, this imparity of estrogen notifies the unusual functionality of the reproductive system, nervous system as well a immune systems of both the human beings and animals(. Although, a lot of previous and recent studies are revealed that the bisphenols are primary agents that the ability to lower the sperms and effect the male sterility and also may defray the female reproductive system and unusual linked to the destroy the linked qualities with respect to health problems including cancer, diabetes,

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metabolic diseases and cardiovascular diseases(13).

5. Conflicts of Interest None.

6. References

[1] Konieczna, A., Rachoń, D., Owczarek, K., Kubica, P., Kowalewska, A., Kudłak, B., … Namieśnik, J.

(2018). Serum bisphenol A concentrations correlate with serum testosterone levels in women with polycystic ovary syndrome. Reproductive Toxicology. doi:10.1016/j.reprotox.2018.09.006

[2] Li, M.-K., Hu, L.-Y., Niu, C.-G., Huang, D.-W., & Zeng, G.-M. (2018). A magnetic separation fluorescent aptasensor for highly sensitive detection of bisphenol A. Sensors and Actuators B:

Chemical, 266, 805–811. doi:10.1016/j.snb.2018.03.163

[3] Zhang, Y.-F., Ren, X.-M., Li, Y.-Y., Yao, X.-F., Li, C.-H., Qin, Z.-F., & Guo, L.-H. (2018). Bisphenol A alternatives bisphenol S and bisphenol F interfere with thyroid hormone signaling pathway in vitro and in vivo. Environmental Pollution, 237, 1072–1079. doi:10.1016/j.envpol.2017.11.027

[4] Konieczna, A., Rachoń, D., Owczarek, K., Kubica, P., Kowalewska, A., Kudłak, B., … Namieśnik, J.

(2018). Serum bisphenol A concentrations correlate with serum testosterone levels in women with polycystic ovary syndrome. Reproductive Toxicology. doi:10.1016/j.reprotox.2018.09.006

[5] Zhou, Q., Jin, Z., Li, J., Wang, B., Wei, X., & Chen, J. (2018). A novel air-assisted liquid-liquid microextraction based on in-situ phase separation for the HPLC determination of bisphenols migration from disposable lunch boxes to contacting water. Talanta, 189, 116–121.

doi:10.1016/j.talanta.2018.06.072

[6] Yao, Q., Feng, Y., Tan, C., Xia, S., Zhao, L., Wang, S., … Chen, X. (2018). An on-line solid-phase extraction disc packed with a phytic acid induced 3D graphene-based foam for the sensitive HPLC- PDA determination of bisphenol A migration in disposable syringes. Talanta, 179, 153–158.

doi:10.1016/j.talanta.2017.11.003

[7] Liu, M., Jia, S., Dong, T., Han, Y., Xue, J., Wanjaya, E. R., & Fang, M. (2019). The occurrence of bisphenol plasticizers in paired dust and urine samples and its association with oxidative stress.

Chemosphere, 216, 472–478. doi:10.1016/j.chemosphere.2018.10.090

[8] Duan, Y., Yao, Y., Wang, B., Han, L., Wang, L., Sun, H., & Chen, L. (2018). Association of urinary concentrations of bisphenols with type 2 diabetes mellitus: A case-control study. Environmental Pollution. doi:10.1016/j.envpol.2018.09.093

[9] Marqueño, A., Pérez-Albaladejo, E., Flores, C., Moyano, E., & Porte, C. (2018). Toxic effects of bisphenol A diglycidyl ether and derivatives in human placental cells. Environmental Pollution.

doi:10.1016/j.envpol.2018.10.045

[10] Arce, M. M., Sanllorente, S., Ortiz, M. C., & Sarabia, L. A. (2018). Easy-to-use procedure to optimise a chromatographic method. Application in the determination of bisphenol-A and phenol in toys by means of liquid chromatography with fluorescence detection. Journal of Chromatography A, 1534, 93–

100. doi:10.1016/j.chroma.2017.12.049

[11] Bolat, G., Yaman, Y. T., & Abaci, S. (2018). Highly sensitive electrochemical assay for Bisphenol A detection based on poly (CTAB)/MWCNTs modified pencil graphite electrodes. Sensors and Actuators B: Chemical, 255, 140–148. doi:10.1016/j.snb.2017.08.001

[12] De Bleye, C., Dumont, E., Hubert, C., Sacré, P.-Y., Netchacovitch, L., Chavez, P.-F., … Ziemons, E.

(2015). A simple approach for ultrasensitive detection of bisphenols by multiplexed surface-enhanced Raman scattering. Analytica Chimica Acta, 888, 118–125. doi:10.1016/j.aca.2015.07.023

[13] Wu, P., Cai, Z., Jin, H., & Tang, Y. (2018). Adsorption mechanisms of five bisphenol analogues on PVC microplastics. Science of The Total Environment. doi:10.1016/j.scitotenv.2018.09.049

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