Tuesday 31 July 2012

Ultra High Performance Liquid Chromatography (UHPLC)




  • Shorter and smaller inside-diameter columns used
  •  2 µm stationary phase
  • System frequently runs at pressures in excess of 600 bar
  • Requires more-sophisticated instrumentation due to very high pressure that the system needs to withstand
  • Use of UHPLC improves productivity and throughput, also reduces solvent consumption
  • UHPLC provides significant advantages in terms of speed, peak resolution, sensitivity and peak capacity.





Friday 6 July 2012

Introduction

Due to emergence of findings like BPA exposure can influence brain development, the bisphenol A(BPA) has become well known in the recent years as heath concerns were raised. The concerns over BPA began with baby bottles and spread to include other types of bottles and children toys.

Bisphenol A (BPA) is an organic compound, having the chemical formula (CH3)2C(C6H4OH). In its pure form, they are white flakes with a faint phenol-like smell.



The compound was first synthesized by the Russian chemist A.P. Dianin in 1905. It is produced by condensation reaction of phenol with acetone in the presence of a strongly acidic ion-exchange resin as catalyst.

File:Synthesis Bisphenol A.svg

Bisphenol A is moderately soluble in water (300 mg/L) and it dissociates in an alkaline environment (pKa 9.9–11.3).  It is used in the sythesis of polycarbonate, epoxy, unsaturated polyester and polysulfone resin. The process of synthesis of polycarbonate resins is shown below.



BPA is also utilised in PVC (polyvinyl chloride) production as a polymerization inhibitor, where after complete polymerization, residual BPA may remain. And as antioxidant in glues, plastics and ink. From 1957 to 2001, global production of BPA was estimated to be an astonish amount of 2.5 million tons.

There are many applications of materials containing BPA. Some common applications are as varnish on the cans' internal surface and as packaging for storage of food, drinks and pharmaceuticals. Uses of epoxy resins include industrial floorings, adhesives, industrial protective coatings, powder coatings, automotive primers and printed circuit boards.

On the other hand, polycarbonates are used in medical equipment (for dialysis and blood oxygenation), water bottles, cell phones,eyeglass lenses, consumer electronics, digital media (e.g. CDs and DVDs) due to its desired properties for both optical clarity and heat resistance.

However, in the production of polycarbonate polymer, not all of the BPA is used up, and they may leach out of the polymer. This is an area of concern as people are afraid that BPA will leach out and be taken in by the human body in one way or another and have adverse effect on health. A possible case is the leaching of BPA from baby bottles composed of polycarbonate into the liquid stored and be consumed by infants. Recently, many applications of polycarbonate have been replaced with new copolymers, such as co-polyester, to eliminate BPA.

Currently, BPA is now monitored in specific products, like baby bottles and other children’s products due to health concerns over exposue to the compound. Simple and rugged analytical methods are needed to determine the presence and amount of BPA in plastic materials. This blog will present the extraction and HPLC analysis of children’s products for BPA with reference from other resources.

Sources:
1.http://www.perkinelmer.com/CMSResources/Images/44-74055app_BPAinToys.pdf
2.http://www.bisphenol-a.org/about/bpa-info/bpa-synthesis.html
3.http://apps.kemi.se/flodessok/floden/kemamne_Eng/bisfenol_a_eng.htm
4.http://en.wikipedia.org/wiki/File:Synthesis_Bisphenol_A.svg
5. http://www.us.edu.pl/uniwersytet/jednostki/wydzialy/chemia/acta/ac16/zrodla/01_AC16.pdf

UHPLC Separation and detection of Bisphenol A (BPA) in Plastics



Experimental

Using UHPLC, BPA was extracted from children toys and analysed. Two different extraction technique was carried out which portrayed situations in which children might come in contact with BPA. 3 different samples (cube, die, and dwarf), 30g each were used for the separation. For the first method, the samples were submerged separately in 1L of water at 40 ˚C for 24 hours. The second method involved the samples being submerged separately in 1L HCL (0.07M) at 37˚C for 2 hours. Using a PerkinElmer Flexar™ FX-10 UHPLC system and a PerkinElmer Series 200a Fluorescence detector, the extracted samples were analysed. Separation of the components in the samples was completed on a Brownlee Validated C8 Column (see Table 1).



The BPA analyzed with the given LC conditions eluted at 5.43 mins (Figure 3). The external standard calibration method was used. The UHPLC system was calibrated across a range of 1 – 50 ppb (µg/L) BPA (Table 2).



 A calibration graph was plotted. BPA in the extracts of the toy samples were quantified using the calibration curve generated during standard analysis (Table 3).
To ensure that the system was free from any contamination of BPA, blanks are injected and analysed between standard and samples.


Results
Figure 4 shows the chromatogram of the water extract of the toy dwarf sample.
Referring to table 3, toy samples which were submerged in water at 40 ˚C for 24 hours showed a much higher amount of BPA than in acid. In all the 3 water extraction from different toy samples, BPA was found within the calibration range of the standard curve.

Conclusion

Analysis in plastics is becoming more important due to health concerns over exposure to BPA. From this experiment, all 3 toy samples have BPA identified in it.


Source: http://www.perkinelmer.com/CMSResources/Images/44-74055app_BPAinToys.pdf

News: U.Va. Research Finds Effects of Common Chemical BPA Span Generations

U.Va. Research Finds Effects of Common Chemical BPA Span Generations

Bisphenol A, or BPA – a chemical that commonly leaches into food and drink from cans and plastic containers – can alter the behavior of mice for generations after exposure, research led by the University of Virginia School of Medicine shows.
The researchers exposed female mice to the compound prior to and during pregnancy, feeding them sufficient doses to create a BPA level in the blood equivalent to what is typically found in humans. The researchers then examined the genetic effects on subsequent generations. They found that the initial BPA exposure continued to affect gene expression and shape social behavior in the fourth generation – the last examined – though there had been no additional exposure to the chemical.
The researchers believe BPA has trans-generational effects in people the same as it does in mice. "Based on our data, and the results from others, it is clear the effects of this chemical are going to be with us for a long, long time," said Emilie F. Rissman, the study's lead investigator and a professor of biochemistry and molecular genetics at the School of Medicine.

BPA is an endocrine disruptor, meaning it interferes with the body's hormone system. The man-made chemical is commonly used in polycarbonate plastics, and it is often found in can linings and food and drink packaging. Because BPA is water-soluble, it can leach into food and drink, especially when the container gets hot or if the food is acidic. This is the primary way people are exposed to BPA.
A survey conducted by the Centers for Disease Control and Prevention in 2003-04 found detectable levels of BPA in 93 percent of 2,517 urine samples taken from Americans age 6 and older.

In examining the trans-generational effects of BPA, the researchers looked at the social interactions between pairs of juvenile mice in the generations after exposure to BPA. They evaluated the amount of time the mice spent exploring their environment and the amount of time they spent engaged in social activities. They also conducted targeted genetic testing to determine BPA's effects on genetic expression patterns in the brain.

researchers found that the mice exposed to BPA while in the womb were less social than mice that had not been exposed. By the third subsequent generation, the behavior had flipped: Mice descended from the BPA-exposed mouse were more social than the control group.

The researchers' conclusion: "Exposure to a low dose of BPA, only during gestation, has immediate and long-lasting, trans-generational effects," they write in a paper published online in the journal Endocrinology.
While the researchers believe BPA may have trans-generational effects in humans as well, the effects may manifest differently.

"While we certainly observed behavioral changes that were passed from generation to generation with subsequent exposure, I cannot say for sure these effects would be the same in humans," Rissman said. "However, as fellow mammals with a 99 percent similarity in their genomes, mice are a good laboratory model for investigations like these, which simply cannot be done in people. While work in humans is correlational, it is important and worth examination."
The researchers next plan to examine the mechanism by which BPA is affecting genes.

Source: http://insciences.org/article.php?article_id=10841

Tuesday 19 June 2012

News: BPA exposure effects may last for generations

Exposure to low doses of Bisphenol A (BPA), a toxic man-made chemical, during gestation has immediate and long-lasting trans-generational effects on the brain and social behaviour in mice, according to a recent study accepted for publication in the journal Endocrinology, a publication of The Endocrine Society.



BPA is a man-made chemical present in a variety of products including food containers, receipt paper and dental sealants and is now widely detected in human urine and blood. Public health concerns have been fueled by findings that BPA exposure can influence brain development. In mice, prenatal exposure to BPA is associated with increased anxiety, aggression and cognitive impairments.

"We have demonstrated for the first time to our knowledge that BPA has trans-generational actions on social behavior and neural expression," said Emilie Rissman, PhD, of the University of Virginia School of Medicine and lead author of the study. "Since exposure to BPA changes social interactions in mice at a dose within the reported human levels, it is possible that this compound has trans-generational actions on human behavior. If we banned BPA tomorrow, pulled all products with BPA in them, and cleaned up all landfills tomorrow it is possible, if the mice data generalize to humans, that we will still have effects of this compound for many generations."

In this study, female mice received chow with or without BPA before mating and throughout gestation. Plasma levels of BPA in supplemented female mice were in a range similar to those measured in humans. Juveniles in the first generation exposed to BPA in utero displayed fewer social interactions as compared with control mice. The changes in genes were most dramatic in the first generation (the offspring of the mice that were exposed to BPA in utero), but some of these gene changes persisted into the fourth generation.

"BPA is a ubiquitous chemical, it is in the air, water, our food, and our bodies," said Rissman. "It is a man-made chemical, and is not naturally occurring in any plant or animal. The fact that it can change gene expression in mice, and that these changes are heritable, is cause for us to be concerned about what this may mean for human health."


Other researchers working on the study include: Jennifer Wolstenholme, Michelle Edwards, Savera Shetty, Jessica Gatewood and Jessica Connelly of the University of Virginia; and Julia Taylor of the University of Missouri.