 Direct Thermal printing is employed in various commercial industries

 Business applications include receipts, luggage tags, labels

 Heat is used instead of ink during the printing process, making this much more cost-
effective
 A printed image will form when thermal coated paper(thermochromic paper) is heated
 Thermochromism – this color change brought about by the temperature
 Thermochromic paper used basically consist of two layers: Base Paper and Thermal
Sensitive layer

 Base Paper – standard paper

 Thermal Sensitive Layer – usually comprise of three components: Solvent, Dye,
Color Developer
 Solvent – long chain aliphatic compound (e.g., fatty acids, amide, alcohol);
Melting Point: 45-65˚C
 Thermochromic Dye – special ink that change color when exposed to
different temperature (usually from colorless to a colored one)
 Crystal Violet Lactone (CVL) is usually used
 Opening up the lactone ring increases conjugation; this conjugation
enables dye to absorb visible light
 Color Developer – usually are weak acids that donate protons to the dye
molecules
 This open up the lactone ring, thus producing a dark color
 BPA is the usual color developer used
 One of the nondietary source of BPA in humans is via dermal absorption
 BPA in thermal receipts are typically in their free form / unbound state
 Because of this, BPA can be readily transferred to the skin upon contact with
it
 Presence of BPA in thermal paper receipts could contribute to the total BPA
exposure (asides from diet, which is the primary exposure culprit)
 In line with this, the highest vulnerable group to this type of BPA
exposure is workers (e.g., cashiers )
 BPA discharge to the environment is generating considerable concern due to its adverse
effects, particularly in humans
 BPA is considered as an Endocrine-disrupting chemical (EDC)
 BPA has a structure that bears resemblance to estradiol
 BPA can mimic estrogen thus this substance can bind with estrogen
receptors
 BPA at low concentration can still has a drastic effect on the human body
 BPA is a suspected contributing agent in in cardiovascular diseases,
diabetes, metabolic disorders, prostate cancer, neurobehavioral and
neuroendocrinal effects, and immune system alterations
 MATERIALS AND METHODS
 Sample Collection
 Proposed Duration of Sample Collection : A month
 Purposive Sampling
 Receipts can be gathered either by soliciting receipts from customers and/or after
purchasing from stores
 After collecting the receipts, these prospect samples will be wrapped with aluminum
foil then stored in a zipped storage bag.

 Identification of Thermal Paper

 Part of the sample receipt will be heated on a hotplate at 200 ˚C. If the paper
darkens, that is an indicator that sample is indeed a thermal paper.
 Thermal paper samples will be wrapped with aluminum foil and stored in a dark
place before analysis.

 Sample Extraction
 Method that will be utilized is based on the work of (Rocha et al., 2015), which
is a modification from the developed model of (Liao & Kannan, 2011)
 A circular spot (12 mm in diameters, 7-8 mg) will be acquired from the middle portion
of the receipts.
 It will be then cut into pieces, placed in a polypropylene conical tube (15 ml).
Methanol (5 ml) will be added.
 Tube will be then shaken in a vortex mixer (60 s) followed by ultra sonification (10
min).
 Centrifugation will then be executed at 4000 x g
 Supernatant (20 μl) will be placed in another tube then diluted with mixture of
Methanol-Water (10 ml, 1:1). This solution will then be spiked with the internal
standard (BPA-d16)
 These solutions will then be stored at -20 ˚C until analysis.
 Before the analysis, aliquots will be filtered through a 0.45 μm filter.
 Each sample will be analyzed in duplicate.

 Instrumentation
 

 Quality Assurance / Quality Control

 Analysis of effectivity of Extraction
 10 samples will be randomly selected
 These samples will undergo extraction and analysis again to determine whether
BPA/BPS have been extracted thoroughly
 If there’s no amount of these chemicals that will be detected after the
reextraction, that will be sufficient evidence that the extraction performed is
efficient

 Blanks
 For each batch of 20 samples, a Procedural blank, a spiked blank, and a
spiked paper blank will also be processed
 Procedural blank – contains water instead of sample; check for interference
and./or contamination
 Spiked blank – mixture of BPA & BPS (50 ng/ml)
 Mean ± SD will be obtained for the recoveries of BPA & BPS
 Relative Standard Deviation (RSD) will be obtained for the replicate analysis

 Calibration Curve
 Ratios of peak area of BPA or BPA-d16 internal standard in Methanol-Water will be
plotted
 The sensitivity of the instrument will be assessed by injecting duplicate samples as
well as evaluating calibration check standard after every ten samples
 Limit of Detection (LOD) and Limit of Quantification (LOQ) will be calculated
using the following formulas:
 LOD = 3 SB/m
 LOQ = 10 SB/m
 SB = Standard deviation of 10 blank determinations
 m = slope of calibration curve

 Analysis
  BPA & BPS confirmation will be done by comparing the retention time of the
chromatographic peak against the standard
 Estimation of Daily Intake/Exposure
 Estimated Daily Intake (EDI) will be determined using this equation:
 EDI = k x C x HF x HT x AF/106
 k = paper-to-skin transfer coefficient of BPA (21,522.4 ng/s)
 C = concentration of BPA in paper samples (μg/g)
 HF = Handling Frequency (times/day)
 2  general population
 150  cashiers
 HT = Handling Time (assumed to be 5 s)
 AF = Absorption Fraction of BPA by skin (27%)
 From the calculated values, the estimated fifth percentile, median and 95th
percentile daily intakes of BPA/BPS can be acquired
 EDI values will also be divided by the average body weight of an adult in order to
measure the intakes adjusted for weight (usually 70 or 80 kg)