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In vitro testing for oral bioaccessibility of

trace metals in Ceylon tea samples


HAPUARACHCHI H.D.H.M
IAC/0040

M.Sc. Industrial Analytical Chemistry Research Supervisors :


Department of Chemistry Dr. Jagath Wanshapala
University of Sri Jayewardenepura Dr. Dilru Rathnaweera
11/1/19
2016 1
CONTENTS
 Introduction
 Objectives
 Methodology
Time Frame
 References
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OBJECTIVES
Objectives

The aim of this study is to examine the oral bioaccessibility


(maximum soluble concentration in gastrointestinal medium) of
total metal (MT) in brewed tea extracts for the following metals:

 Lead
 Iron
 Cadmium

 Manganese
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 Zinc
INTRODUCTION
• Tea is one of the most popular nonalcoholic beverages
in the world. As tea contains essential nutrients
consuming tea is beneficial for human health.

• Intake of food contaminated by heavy metals is harmful


to human health. Various studies have discussed the
potential health implications of trace metals in tea.

• Particularly since the tea bush is known to accumulate


trace metals (Bosque et al., 1990, Anonymous, 1999)

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• The main sources of heavy metals in plants are their
media, nutrients, agro inputs and soil. Other sources
may include pesticides and fertilizers.

• Elevated heavy metal levels cause damage to plants


such as delayed flowering, lower chlorophyll content
and reduction in the number and quality of shoots
(Setia, Kaur, & Setia,1989).

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• In-vitro gastrointestinal extraction, also known as
bio accessibility, is important when assessing
chemical risk to humans.

• Bioaccessibility is defined as the proportion of total


metal in food for absorption.

• In use, it purports to simulate the release of


chemicals from sample matrices (e.g., food and soil)
that may be consumed intentionally or
unintentionally in the diet, so in-vitro conditions are
created.
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SRI LANKA TEA BOARD GUIDELINES (2010)

Heavy Metals:

• Cadmium - max 0.2 ppm


• Lead - max 5ppm
• Copper - max 100 ppm
• Zinc - max 100 ppm
• Iron - max 500 ppm

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MATERIALS & METHOD
INSTRUMENTS
• Flame Atomic Absorption Spectrometer (FAAS)
• Centrifuge
• Muffle furnace
• Microwave assisted digestion unit
• Mechanical shaker

REAGENTS
• Deionized water
• Standard Metal Solutions
• Enzymes and bile salts (Pepsin, Pancreatin, Bile extract)
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SAMPLE COLLECTION & PREPARATION
• Orthodox (Whole leaf )and CTC (Crush ,Tear, Curl)processed
tea samples will be collect from following major tea producing
areas according to Tea Market Update volume 12 ,No 2
Galle
Badulla
Matara
Kandy
• Organically grown Eliya
Nuwara tea sample will be collect as a control sample
• Soil andwater samples from cultivations will be collected random
Rathnapura
manner for heavy metal analysis

• 2.0 g of tea sample will brew with 150 mL of 100°C distilled


11/1/19 deionised water for 5 minutes 9
11/1/19 Minekus, M., Alminger, M., Alvito,et. al (2014) 10
IN VITRO GASTRO INTESTINAL DIGESTION
In vitro gastro intestinal method – Marisa Intawongse, Jhon R. Dean, Treands in Analytical chemistry, Vol.25, No.9, 2006

ORAL PHASE GASTRIC PHASE INTESTINAL PHASE


• pH = 6.5 • pH = 2.0 • pH=5.0
• Saliva • Pepsin • Pancreatin
• Mechanical shaking • 37°C for 2 h • Bile extract
• 37°C for 2h

• Centrifuge the digested samples at 15000 rpm at 4°C for


30 minutes

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Total Metal concentration Total Metal Concentration
in tea extract in bioaccessible fraction
Supernatant and
Untreated tea extract residue will be
will be wet ashed with separated and wet
5 mL of 50% HNO3 ashed with 5 mL of
50% HNO3

• Metal concentration of treated and untreated samples


quantified using FAAS

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STATISTICALStatistical
ANALYSIS Analysis

• Correlation coefficient will be calculated


between total metal concentration and
bioaccessible fraction of metal in tea samples.
(p<0.05)

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TIME FRAME

Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep

Literature survey

Experimental work

Thesis writing

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REFERENCES References
• Minekus, M., Alminger, M., Alvito, P.,et. al (2014) A standardised static in vitro digestion method
suitable for food - an international consensus. Food & Function. 5 (6) 113-1124.
• Falahi, E. and Hedaiati, R. (2013). Heavy metal content of black teas consumed in Iran. Food Additives
and Contaminants: Part B, 6(2), pp.123-126.
• Harmanescu, M., Alda, L., Bordean, D., Gogoasa, I. and Gergen, I. (2011). Heavy metals health risk
assessment for population via consumption of vegetables grown in old mining area; a case study:
Banat County, Romania. Chemistry Central Journal, 5(1), p.64.
• Ho, C., Lin, J. and Shahidi, F. (2009). Tea and tea products. Boca Raton: CRC Press.
• Seenivasan, S., Manikandan, N., Muraleedharan, N. and Selvasundaram, R. (2008). Heavy metal
content of black teas from south India. Food Control, 19(8), pp.746-749.
• Soylak, M., Tuzen, M., Souza, A., Korn, M. and Ferreira, S. (2007). Optimization of microwave assisted
digestion procedure for the determination of zinc, copper and nickel in tea samples employing flame
atomic absorption spectrometry. Journal of Hazardous Materials, 149(2), pp.264-268.
• J.M Laparra, D. Velez et.al (2003)Estimation of Arsenic Bioaccessibility in edible Seaweed by an invitro
digestion Method: J.Agric.Food che, pp51

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THANKYOU!!!

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