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