Application of High-Performance Liquid Chromatography Combined with Fluorescence Detector and Dispersive Liquid–Liquid Microextraction to Quantification of Selected Bisphenols in Human Amniotic Fluid Samples
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bisphenol Standards
2.2. Chemicals Used during the Instrumental Analysis and DLLME Procedure and Laboratory Equipment
2.3. Instrumental Analysis
2.4. Method Validation
2.5. Selectivity
2.6. Linearity
2.7. Extraction Recovery Studies
2.8. Optimized DLLME Procedure for Amniotic Fluid Samples
2.9. Human Amniotic Fluid Sample Collection and Storage
3. Results
3.1. Optimizing the Procedure
3.2. Recovery Studies
3.3. Chromatographic Conditions
3.4. Detection Conditions
3.5. Optimization of the HPLC-FLD Procedure for Quantitative Analysis
3.6. Applying the Procedure for Sample Spiked with Amount Close to Real Concentration Found in Human Body Fluids
4. Discussion
- -
- The sample volume stayed the same (0.4 mL);
- -
- The method was optimized for 5 bisphenols, while the previous study utilized a method optimized for 11 bisphenols. However, in the previous study, some bisphenols were not fully separated from each other (for example, BPAP and BPAF). For recovery studies, mixtures of 11 bisphenols were divided in two parts;
- -
- The SPE procedure was more time- and solvent-consuming;
- -
- Recoveries and repeatability remained at acceptable levels;
- -
- The LOQ values were 3.2–12.6 ng/mL for [33] and 6.17–22.72 ng/mL for this work.
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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No. | Bisphenol | IUPAC Name 1 | Chemical Structure | Log p 1 | pKa 1 |
---|---|---|---|---|---|
1 | BPF | 4-[(4-hydroxyphenyl)methyl]phenol | 2.9 | pKa1 = 7.55 pKa2 = 10.80 | |
2 | BPE | 4-[1-(4-hydroxyphenyl)ethyl]phenol | 3.9 | - | |
3 | BPAF | 4-[1,1,1,3,3,3-hexafluoro-2-(4-hydroxyphenyl)propan-2-yl]phenol | 4.5 | 9.2 | |
4 | BADGE∙ 2HCl | 1-chloro-3-[4-[2-[4-(3-chloro-2-hydroxypropoxy)phenyl]propan-2-yl]phenoxy]propan-2-ol | 4.6 | - | |
5 | BPP | 4-[2-[4-[2-(4-hydroxyphenyl)propan-2-yl]phenyl]propan-2-yl]phenol | 6.1 | - |
Bisphenol | 3 mL of Acetone: Dichloromethane (2:1, v/v) | 3 mL of Acetone: Chloroform (2:1, v/v) | 3 mL of Acetone: Dichloromethane: Chloroform (4:1:1, v/v/v) | |||
---|---|---|---|---|---|---|
Recovery | RSD% | Recovery | RSD% | Recovery | RSD% | |
BPF | 82.6% | 11% | 87.9% | 9% | 84.2% | 9% |
BPE | 81.7% | 8% | 94.3% | 8% | 88.8% | 10% |
BPAF | 107.2% | 9% | 105.5% | 11% | 97.6% | 7% |
BADGE∙2HCl | 101.4% | 10% | 103.9% | 5% | 99.7% | 16% |
BPP | 100.8% | 14% | 106.7% | 11% | 102.3% | 13% |
No. | Bisphenol | Recovery Values (%) ± RSD% for Spiking Level (ng/mL) | |||
---|---|---|---|---|---|
20 | 30 | 40 | 50 | ||
1 | BPF | 104.9 ± 9 | 98.9 ± 9 | 109.2 ± 9 | 87.9 ± 9 |
2 | BPE | 115.9 ± 6 | 105.1 ± 5 | 105.1 ± 5 | 94.3 ± 8 |
3 | BPAF | 113.3 ± 7 | 97.0 ± 6 | 96.2 ± 7 | 106.7 ± 11 |
4 | BADGE∙2HCl | 80.9 ± 6 | 98.9 ± 5 | 109.2 ± 7 | 87.9 ± 5 |
5 | BPP | 99.0 ± 5 | 105.1 ± 12 | 105.1 ± 6 | 94.3 ± 11 |
No. | Bp. | Retention Time, tR (min) | Linear Regression | Standard Deviation (SD) of Slope | SD of Intercept | Coefficient of Determination (R2) | LOD (ng/mL) | LOQ (ng/mL) | Confidence Level |
---|---|---|---|---|---|---|---|---|---|
1 | BPF | ~10.13 | y = 5.875x − 5.675 | 0.31 | 7.66 | 0.9916 | 4.30 | 13.04 | 0.95 |
2 | BPE | ~11.27 | y = 6.0876x + 3.7012 | 0.24 | 5.95 | 0.9953 | 3.23 | 9.77 | 0.95 |
3 | BPAF | ~15.38 | y = 14.956x − 12.788 | 1.38 | 33.99 | 0.9816 | 7.50 | 22.72 | 0.95 |
4 | BADGE∙2HCl | ~18.15 | y = 16.592x + 86.834 | 0.14 | 3.37 | 0.9998 | 2.04 | 6.17 | 0.95 |
5 | BPP | ~19.79 | y = 9.2674x + 471.42 | 0.42 | 10.38 | 0.9938 | 3.70 | 11.20 | 0.95 |
Analyzed Bisphenols | Sample Type | Instrumental Analysis | Sample Volume | Extracting Mixture (Dispersive Solvent + Extracting Solvent) | LOD/LOQ | Ref. |
---|---|---|---|---|---|---|
BPA, BPS, BPF, BPZ, BPP, BPAF, BPAP | Urine | LC-MS/MS | 5 mL of urine + 5 of mL water | 750 µL of acetone + 500 µL of 1,2-dichloroethane | LOQ: 0.03–0.2 ng/mL | [27] |
BPA, BPF, BPZ, BP | Urine | GC-MS | 2 mL of urine + 8 of mL water | 1 mL of acetone + 100 µL chloroform | LOD: 0.01–0.04 ng/mL | [28] |
BPA, BPS, BPP, BPAP, BPAF, BPZ | Saliva | LC-MS/MS | 500 µL of saliva + 500 µL of water | 1.5 mL of acetone + 500 µL of chloroform | LOQ: 0.05–0.4 ng/mL | [29] |
BPF, BPE, BPA, BPB, BPAF, BADGE∙2HCl, BPP | Amniotic fluid collected during amniocentesis | HPLC-FLD | 400 µL of amniotic fluid + 1.6 mL of water | 2 mL of acetone + 1 mL of chloroform | LOD: 2.04–7.5 ng/mL | This work |
Bisphenol | Sample Amount | Extraction Technique Used during Sample Preparation | Chromatographic Method and Detection Technique | LOD/LOQ | Ref. |
---|---|---|---|---|---|
BPA | 0.5 mL | SPE (cartridges packed with 200 mg of silica-based bonded C18 material) | LC-MS | LOD = 0.1 ng/mL LOQ = 0.3 ng/mL | [36] |
BPA | 1.0 mL | SPE (C18-based sorbent) | GC-MS | LOD = 0.52 ng/mL | [37] |
BPA, BPP, BPS, BPAF, BPAP | 2.0 mL | Solvent extraction (ethyl acetate) | LC-MS | LOQ: from 0.01 ng/mL to 0.2 ng/mL | [38] |
BPF, BPE, BPAF, BADGE∙2HCl, BPP | 0.4 mL | DLLME | HPLC-FLD | LOQ: from 6.17 to 22.72 ng/mL | This work |
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Szubartowski, S.; Tuzimski, T. Application of High-Performance Liquid Chromatography Combined with Fluorescence Detector and Dispersive Liquid–Liquid Microextraction to Quantification of Selected Bisphenols in Human Amniotic Fluid Samples. Int. J. Environ. Res. Public Health 2023, 20, 297. https://doi.org/10.3390/ijerph20010297
Szubartowski S, Tuzimski T. Application of High-Performance Liquid Chromatography Combined with Fluorescence Detector and Dispersive Liquid–Liquid Microextraction to Quantification of Selected Bisphenols in Human Amniotic Fluid Samples. International Journal of Environmental Research and Public Health. 2023; 20(1):297. https://doi.org/10.3390/ijerph20010297
Chicago/Turabian StyleSzubartowski, Szymon, and Tomasz Tuzimski. 2023. "Application of High-Performance Liquid Chromatography Combined with Fluorescence Detector and Dispersive Liquid–Liquid Microextraction to Quantification of Selected Bisphenols in Human Amniotic Fluid Samples" International Journal of Environmental Research and Public Health 20, no. 1: 297. https://doi.org/10.3390/ijerph20010297