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SIMULTANEOUS DETERMINATION OF BISPHENOL A AND BISPHENOL S
LEACHING FROM PLASTICS USING UV/VIS SPECTROPHOTOMETRY AND
HPLCAnalytical Chemistry Research performed by
Jean Eudes BenecyoUnder supervision of Dr. Sara Hubbard
Dr. J. D. Patterson Summer Research ProgramOuachita Baptist University
https://www.niehs.nih.gov/health/topics/agents/sya-bpa/#a739817
Exposure to BPA and BPS
Bisphenol A & Bisphenol S■ Plasticizer■ Synthetic estrogen■ Endocrine disruptor that
mimics estrogen■ It affects the brain, behavior,
and the prostate gland in fetuses, infants, and children
■ Plasticizer■ First made in 1869 as a dye■ Endocrine disruptor■ After research linked
Bisphenol A to adverse health effects, it was substituted by Bisphenol S
Bisphenol SBisphenol A Estrogen
Objectives
■ Develop a method of simultaneous determination of BPA and BPS using– UV/VIS Spectrophotometry– HPLC
■ Apply these methods in real world samples
UV/VIS Spectrophotometry of mixtures
Detection
Vernier UV-VIS Spectrophotometer with Logger Pro 3 software
A+B solution
X YλX λY
A’A’’
X+Y
A’ = ε‘X.b.[X] + ε‘Y.b.[Y]A’’ = ε‘’X.b.[X] + ε‘’Y.b.[Y]
UV/VIS Spectrophotometry
A’ = ε‘BPA.b.[BPA] + ε‘BPS.b.[BPS]A’’ = ε‘’BPA.b.[BPA] + ε‘’BPS.b.[BPS]• Where A: Absorbance; ε: Molar Absorptivity; b: Cell
size (b = 1cm); • [ ]: Concentration in μg/mL; • ‘ : at 255.1nm; ‘’ : at 272.3nm.
Method Development
■ Fig 1. UV/VIS Absorbance calibration curves of BPA.
■ LODBPA at 255.1nm= 1.18μg/mL; LODBPA at 272.3nm= 0.48μg/mL; LOQBPA at 255.1nm= 3.93μg/mL; LOQBPA at 272.3nm= 1.60μg/mL.
■ Fig 2. UV/VIS Absorbance calibration curves of BPS.
■ LODBPS at 255.1nm= 0.06μg/mL; LODBPS at 272.3nm= 0.13μg/mL; LOQBPS at 255.1nm= 0.21μg/mL; LOQBPS at 272.3nm= 0.43μg/mL.
BPA BPS
Method Development (cont.)
■ Fig 3. 3.2 μg/mL BPA/BPS UV-VIS Absorbance Spectra in 1:1 methanol/water.
BPSpeak at 255.1nm. BPApeak at 272.3nm.
■ Fig 4. HPLC calibration curves for BPA and BPS in 1:1 methanol/water
255nm
273nm
High Pressure Liquid Chromatography
■ Fig 5. HPLC Separation of BPA and BPS in 1:1 methanol/water.
■ [BPA] = [BPS] = 3.2 μg/mL
The HPLC method for BPA/BPS Analysis was developed by – Injecting 100 μL into Waters Nova-Pak C 18 Column 3.9x150 mm 60 Å 4μm with
a mobile phase of water/acetonitrile (60:40) and a flow rate of 1.0 mL/min
–Detector: UV (230 nm)–Carrier Gas: Helium, 20 mL/min (non-continuous)
■ Table I. Comparison of actual and calculated [BPA] and [BPS] values and determination of the effectiveness of the UV-VIS Absorption Spectrophotometry method by statistical analysis in comparison to the HPLC method
CALCULATEDCONC , μg/mL SD CONC , μg/mL
#1 BPA 1.49 0.134 1.60 8.40 6.60 NOT SSD - -BPS 1.47 0.033 1.60 2.07 7.97 SSD - -
#2 BPA 2.54 0.195 2.40 8.14 -6.01 NOT SSD - -BPS 2.13 0.049 2.40 2.03 11.20 SSD - -
#3 BPA 4.60 0.375 4.80 7.82 4.25 NOT SSD - -BPS 4.15 0.094 4.80 1.97 13.51 SSD - -
#4 BPA 6.41 0.739 9.60 7.70 33.24 SSD - -BPS 8.80 0.197 9.60 2.05 8.38 SSD - -
#5 BPA 2.65 0.281 3.20 8.79 17.26 NOT SSD 2.94 8.13BPS 2.93 0.067 3.20 2.11 8.45 SSD 3.13 2.19
#6 BPA 85.07 2.412 97.90 2.46 13.11 SSD - -BPS 10.04 0.299 10.10 2.96 0.59 NOT SSD - -
#7 BPA 58.02 1.864 65.30 2.85 11.16 SSD - -BPS 11.78 0.304 12.60 2.41 6.47 NOT SSD - -
#8 BPA 9.82 0.757 11.20 6.76 12.30 NOT SSD - -BPS 8.48 0.192 8.81 2.18 3.70 NOT SSD - -
#9' BPA 95.22 2.481 114.00 2.18 16.48 SSD - -BPS 8.75 0.286 8.40 3.41 -4.13 NOT SSD - -
% ERROR
T test result
% ERROR SAMPLE
CALCULATED PREPARED CONC , μg/mL
% RSD CONC
Experimental samples
P1:Baby bottle
(270mL)
P2:Baby bottle
(270mL)
P3:BPA free
Water bottle (500mL)
P4:BPA free
water bottle (500mL)
P5:Nalgene
water bottle (500mL)
P6:Cup
P7:Cup
P8:Party Cup
P9:Luv’n care
Item 10:Thermal
receipt paper
Item 12:Amy’s Chili (Non-BPA
lining)
Item 13:Great value
mixed vegetables
P14:Rubbermaid –
Take along
P15:Nordic ware –
Everyday bowl
P16:Fork and
spoon
Method Application
Summary and Conclusions
UV-VIS Absorption spectrophotometry is effective at simultaneously determining BPA and BPS
Analytical figures of merit were determined for absorption analysis of BPA and BPS
• Low LOD and high linear range
Future Work
Additional analysis of known samples to optimize methods
Determine leaching of BPA/BPS from plastics Use this method in vitro and in vivo
Acknowledgements
Dr. J. D. Patterson Summer Research Program
Dr. Joe Bradshaw