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A P P L I C AT I O N                not e


                                                                                 Liquid Chromatography


                                                                                   Author


                                                                                   Roberto Troiano, PerkinElmer
                                                                                   William Goodman, PerkinElmer




UHPLC separation
and detection of                               Introduction

Bisphenol A (BPA)                              The BPA or bisphenol A (Figure 1) has become well know
                                               over the past year as concerns for its effect on human health
in plastics                                    and well being have been raised. The concerns over BPA
                                               began with baby bottles and spread to include other types of
                                               bottles.
   BPA is used in the production of two very common polymers PVC and Polycarbonate. PVC,
   Polyvinyl chloride, is used in many different products including building materials, medical devices
   and children’s toys. BPA is used in PVC production as a polymerization inhibitor, residual BPA may
   remain after the polymerization is complete. Polycarbonate is another very commonly used plastic.
   It has very desirable properties for both optical clarity and heat resistance. BPA is an important
   monomer in the production of polycarbonate polymer, not all of the BPA is consumed in the pro-
   duction and may leach out of the polymer. Recently, many applications of polycarbonate have been
   replaced with new copolymers, such as co-polyester, to eliminate BPA.




   Figure 1: Structure of Bisphenol A (BPA).
As a result of the health concerns over human exposure to BPA                  Results
    this molecule is now monitored in specific products, including                 The BPA analyzed with the given LC conditions eluted at 5.43
    baby bottles and other children’s products. Simple and robust                  mins (Figure 3). The UHPLC system was calibrated across a
    test methods are needed to determine the presence and amount                   range of 1 – 50 ppb (µg/L) BPA (Table 2).
    of BPA in plastic materials. This paper will present the extraction
    and HPLC analysis of children’s products for BPA.




     Figure 2: Children's toy samples analyzed for BPA in this application note.


    Experimental
    The study presented here includes extraction of BPA from a toy
    matrix and analysis with UHPLC. The extraction procedure
                                                                                          Figure 3: BPA calibration standard at 1 ppb.
    used here is intended to simulate the contact routes through
    which children are likely to encounter BPA. Two different extrac-
    tion techniques were used to analyze BPA in samples (30 g sam-                  Table 2: Table for the analysis of BPA across the range of 1 – 50 ppb (µg/L).

    ple used for each extraction). The first extraction method                      Concentration	                   Response	
    immersed the sample in 1 L of water, at 40 ˚C for 24 hours (EN
    14372). The second immersed the sample with 1 L HCl (0.07                       1 ppb	                           54163	
    M) at                                                                           10 ppb	                          378051	
    37 ˚C for 2 hours. Following extraction the samples were ana-                   20 ppb	                          820335	
    lyzed with a PerkinElmer Flexar™ FX-10 UHPLC system includ-
    ing a PerkinElmer Series 200a Fluorescence detector. The sepa-                  40 ppb	                          1548750	
    ration was performed on a Brownlee Validated C8 Column (see                     50 ppb	                          1957851	
    Table 1).
                                                                                    	r2                              0.9993	
    Table 1: HPLC Conditions for the Analysis of BPA

    HPLC System	                      PerkinElmer Flexar FX-10 UHPLC 	             The limit of quantitation (LOQ) for BPA with the method pre-
    Injection Volume	                 50 μL	                                       sented here is 1 ppb. The signal to noise at the LOQ is approxi-
                                                                                   mately 10:1. The response across the calibration range fit a linear
    Column	                           PerkinElmer C8 (150 mm x 4.6 mm, 5 μm)
                                                                                   calibration with an r2 value of 0.9993. Blanks analyzed between
    Mobil Phase	                      Methanol/Water (65/35)	                      standards and samples showed the system was free from any BPA
    Flow Rate	                        1 mL/min	                                    contamination or carryover.
    Detector Wavelength	              Excitation – 275 nm / Emission – 313 nm      BPA in the extracts of the toy samples were quantified using the
                                                                                   calibration curve generated during standard analysis (Table 3).
    Detector Response Time	 0.1 sec	
                                                                                   Figure 4 shows the chromatogram of the water extract of the toy
    PMT, Em BDW	                      Super High, Wide	                            dwarf sample.
    Run Time	                         15 min	
                                                                                    Table 3: Results from toy sample analysis.
                                                                                    Sample	                   Extraction Type	           µg/L	           µg/g

                                                                                    Cube	                     water	                     2.04	           0.068	
                                                                                    Cube	                     HCl	                       ND	             ND	
                                                                                    Die	                      water	                     3.35	           0.111	
                                                                                    Die	                      HCl	                       1.56	           0.052	
                                                                                    Dwarf	                    water	                     4.32	           0.144	
                                                                                    Dwarf	                    HCl	                       1.78	           0.059	


2
Figure 4: Analysis of toy dwarf for BPA using water.


The extraction procedure which heated the toy for 24 hours in
water at 40 ˚C extracted a significantly higher amount of BPA
from the matrix than the extraction in acid. BPA was found in all
three water extractions within the calibration range of the stan-
dard curve.

Conclusion
As health concerns over exposure to BPA are raised, its analysis
in plastics is becoming very important. The PerkinElmer Flexar
FX-10 UHPLC system provides a sensitive and robust platform
for this analysis. Demonstrated here was a calibration of BPA
across a range of 1 – 50 ppb with a chromatographic run time of
less than 10 minutes. This analysis was applied to 3 toy samples
and BPA was identified in each sample.




PerkinElmer, Inc.
940 Winter Street
Waltham, MA 02451 USA	
P: (800) 762-4000 or
(+1) 203-925-4602
www.perkinelmer.com



For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs

Copyright ©2009, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners.

008731_01	              Printed in USA

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Application Note: UHPLC Separation and Detection of Bisphenol A (BPA) in Plastics

  • 1. A P P L I C AT I O N not e Liquid Chromatography Author Roberto Troiano, PerkinElmer William Goodman, PerkinElmer UHPLC separation and detection of Introduction Bisphenol A (BPA) The BPA or bisphenol A (Figure 1) has become well know over the past year as concerns for its effect on human health in plastics and well being have been raised. The concerns over BPA began with baby bottles and spread to include other types of bottles. BPA is used in the production of two very common polymers PVC and Polycarbonate. PVC, Polyvinyl chloride, is used in many different products including building materials, medical devices and children’s toys. BPA is used in PVC production as a polymerization inhibitor, residual BPA may remain after the polymerization is complete. Polycarbonate is another very commonly used plastic. It has very desirable properties for both optical clarity and heat resistance. BPA is an important monomer in the production of polycarbonate polymer, not all of the BPA is consumed in the pro- duction and may leach out of the polymer. Recently, many applications of polycarbonate have been replaced with new copolymers, such as co-polyester, to eliminate BPA. Figure 1: Structure of Bisphenol A (BPA).
  • 2. As a result of the health concerns over human exposure to BPA Results this molecule is now monitored in specific products, including The BPA analyzed with the given LC conditions eluted at 5.43 baby bottles and other children’s products. Simple and robust mins (Figure 3). The UHPLC system was calibrated across a test methods are needed to determine the presence and amount range of 1 – 50 ppb (µg/L) BPA (Table 2). of BPA in plastic materials. This paper will present the extraction and HPLC analysis of children’s products for BPA. Figure 2: Children's toy samples analyzed for BPA in this application note. Experimental The study presented here includes extraction of BPA from a toy matrix and analysis with UHPLC. The extraction procedure Figure 3: BPA calibration standard at 1 ppb. used here is intended to simulate the contact routes through which children are likely to encounter BPA. Two different extrac- tion techniques were used to analyze BPA in samples (30 g sam- Table 2: Table for the analysis of BPA across the range of 1 – 50 ppb (µg/L). ple used for each extraction). The first extraction method Concentration Response immersed the sample in 1 L of water, at 40 ˚C for 24 hours (EN 14372). The second immersed the sample with 1 L HCl (0.07 1 ppb 54163 M) at 10 ppb 378051 37 ˚C for 2 hours. Following extraction the samples were ana- 20 ppb 820335 lyzed with a PerkinElmer Flexar™ FX-10 UHPLC system includ- ing a PerkinElmer Series 200a Fluorescence detector. The sepa- 40 ppb 1548750 ration was performed on a Brownlee Validated C8 Column (see 50 ppb 1957851 Table 1). r2 0.9993 Table 1: HPLC Conditions for the Analysis of BPA HPLC System PerkinElmer Flexar FX-10 UHPLC The limit of quantitation (LOQ) for BPA with the method pre- Injection Volume 50 μL sented here is 1 ppb. The signal to noise at the LOQ is approxi- mately 10:1. The response across the calibration range fit a linear Column PerkinElmer C8 (150 mm x 4.6 mm, 5 μm) calibration with an r2 value of 0.9993. Blanks analyzed between Mobil Phase Methanol/Water (65/35) standards and samples showed the system was free from any BPA Flow Rate 1 mL/min contamination or carryover. Detector Wavelength Excitation – 275 nm / Emission – 313 nm BPA in the extracts of the toy samples were quantified using the calibration curve generated during standard analysis (Table 3). Detector Response Time 0.1 sec Figure 4 shows the chromatogram of the water extract of the toy PMT, Em BDW Super High, Wide dwarf sample. Run Time 15 min Table 3: Results from toy sample analysis. Sample Extraction Type µg/L µg/g Cube water 2.04 0.068 Cube HCl ND ND Die water 3.35 0.111 Die HCl 1.56 0.052 Dwarf water 4.32 0.144 Dwarf HCl 1.78 0.059 2
  • 3. Figure 4: Analysis of toy dwarf for BPA using water. The extraction procedure which heated the toy for 24 hours in water at 40 ˚C extracted a significantly higher amount of BPA from the matrix than the extraction in acid. BPA was found in all three water extractions within the calibration range of the stan- dard curve. Conclusion As health concerns over exposure to BPA are raised, its analysis in plastics is becoming very important. The PerkinElmer Flexar FX-10 UHPLC system provides a sensitive and robust platform for this analysis. Demonstrated here was a calibration of BPA across a range of 1 – 50 ppb with a chromatographic run time of less than 10 minutes. This analysis was applied to 3 toy samples and BPA was identified in each sample. PerkinElmer, Inc. 940 Winter Street Waltham, MA 02451 USA P: (800) 762-4000 or (+1) 203-925-4602 www.perkinelmer.com For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs Copyright ©2009, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners. 008731_01 Printed in USA