Biology HL's Sample Internal Assessment

Biology HL's Sample Internal Assessment

Effect of varying amounts of burnt Boswellia Frankincense stick gases on refractive index (%) of sugar in sapodilla

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Table of content

Research question

How does the amount of Boswellia serrata burnt (0g, 5g, 10g, 15g, 20g, 25g) affect the refractive index (%) of sugar and the mass of Manilkara zapota (chikoo)  at room temperature?

Rationale

I had taken a road trip to rural Maharashtra, a journey filled with hills and farmland. We stopped at one of the farms, to buy fresh produce, and I got to speaking with the farmer, who told me that vendors use agarbati (Indian frankincense sticks) to ripen produce. This peaked my interest, as I found it unusual. After some reading, I came across a few articles which supported the farmer’s claim. When fruits ripen organically, they release ethylene gas, which is proven to improve their ripening(India). As ethylene is a natural hormone, it does not cause any harm to the health of the consumers of the fruit. With further research I discovered that ethylene was released by the burning of Indian frankincense (Boswellia serrata), which is the main ingredient of agarbatti, also known as loban (Nyanjage et al.) Hence, I started to research on the differences between the natural and artificial process of ripening.  My research also led me to several articles on benefits and disadvantages of artificial ripening.  It led me to ask questions like what exactly happens in the process of fruit ripening and whether we can ensure exactly the right level of fruit ripening for human consumption.

Background knowledge

Ripening of fruits

Ripening is a natural physiological process that makes the fruit sweeter, more palatable, edible, nutritious, softer and attractive. Ripening is also associated with color change due to the pigments that are already present or are produced during ripening (Vaviya et al.) If the ripening process is allowed to continue, the fruit reaches senescence i.e. it becomes overly ripe and now it’s only aim is seed dispersal. It is a natural process and can be slowed down or speeded up, but cannot be completely inhibited. Ethylene gas helps in this process.

Effect of incense sticks on ripening of fruits

Incense sticks produce Ethylene gas on burning. (Marrero et al.) Ethylene is also a gaseous phytohormone produced by plants to carry out both processes of growth and senescence.  It is released naturally to promote fruit ripening (Iqbal et al.). The ripening of fruits is a unique coordination of various biochemical and developmental pathways regulated by ethylene, which affects color, texture, nutritional quality and aroma of fruits. During ripening in climacteric fruits, the ethylene regulates firmness and color changes involving chlorophyll reduction, increase in carotenoids or anthocyanins, sugars, and biosynthesis of volatile organic compounds (Iqbal et al., “Ethylene Role in Plant Growth, Development and Senescence”). Hence it can be theorized that incense sticks will promote artificial ripening of fruits due to presence of ethylene.

Measuring the ripening of fruits using refractive index

In optics, the refractive index or index of refraction n of a material is a dimensionless number that describes how light propagates through that medium. It is defined as:

 

\(n=\frac{ c}{v}\)

 

where, c is the speed of light in vacuum and v is the phase velocity of light in the medium. For example, the refractive index of water is 1.333, meaning that light travels 1.333 times faster in a vacuum than it does in water.

 

The refractive index of a liquid can be measured using a hand refractometer ( a handheld device that works on the critical angle principle by which lenses and prisms project a shadow line onto a small glass reticle inside the instrument, which is then viewed by the user through a magnifying eyepiece. It can be digital or manual) As the concentration of dissolved solids in the liquid changes, the refractive index changes. Increasing concentration of dissolved solids increases the amount of refraction while the amount of refraction decreases with decrease in dissolved solids. In case of increase in process of fruit ripening, an increase in the concentration of sugars leads to an increase in the percentage refractive index. It is this property which is used in many food industries for checking the quality of food products. We shall be using this property for checking amount of sugar dissolved in the extract. (Contento et al.)

Measuring the ripening of fruits using its change in mass

According to research, a fruit will increase in mass as long as sugar is still being transported into it by the plant.  This is possible only as long as the fruit is attached to the main plant i.e. prior to harvesting.  A climacteric fruit is one which can be harvested raw and then allowed to ripen.  This increases the shelf life of the fruit. However, once the fruit is harvested, no further sugar reserves enter the fruit and the change in mass is generally shown as a decline in mass. (Prevention of Post-Harvest Food Losses Fruits.)

 

The decrease in weight of the climacteric fruits on ripening and its ripening is because of three reasons (a) loss of water due to evaporation from surface (b) life processes like respiration being continuously carried out in the plant which reduces the dry matter content in fruit for energy  and (c) hydrolysis of starch into sugar by amylase enzyme.  The greater the decrease in mass, the greater is the ripening of the fruit.

Hypothesis

Experimental Hypothesis

  • The percentage refractive index of sugar in Manilkara zapota extract will increase as the amount of Boswellia serrata burnt increases at room temperature.

This is reasoned as ethylene gas is released by burning Boswellia serrata which is a gaseous hormone that hastens ripening in fruits.

  • The mass of the fruit (Manilkara zapota) extract will decrease as the amount of Boswellia serrata burnt increases at room temperature.

This is reasoned as the fruit has already been harvested and the biochemical changes and water loss can cause reduction in mass.

Null Hypothesis

  • The percentage of refractive index of sugar in Manilkara zapota extract will not show any significant difference as the amount of Boswellia serrata burnt increases at room temperature.
  • 2) The mass of the fruit (Manilkara zapota) extract will not show any significant difference as the amount of Boswellia serrata burnt increases at room temperature.

Variables

Independent VariableHow it was varied?
Mass of Boswellia serrata burnt in gDifferent amounts of Dhoop ( 0g, 5g, 10g, 15g, 20g, 25g) were used in each chamber containing a fixed number of sapodilla

Figure 1 - Table On Independent Variable

Dependent VariableHow it was measured?
Refractive index of fruit extractThe chikoo extract at the end of each experiment were subjected to a refractive index test (using a hand refractometer ) to test for amount of sugar formed.
Change in mass of fruitThe initial and final readings of mass of each fruit were recorded using a digital mass balance.

Figure 2 - Table On Dependent Variable

Control VariableJustification of controlling itMethodology of control
Variety of sapodillaDifferent varieties of sapodilla can have different percentage of ripening capacity i.e. some may ripen in a shorter period of time than others.Using same variety of sapodilla sourced from the same farm. The fruits were collected from a farm from a known farmer who was able to guarantee that they were all picked from the same tree hence were the same variety.
Ripeness of sapodillaInitial level of rawness or ripeness of the fruit may vary, thus affecting the consistency of results.This error in reading can be overcome by taking a large number of samples, so that the average reading eliminates the error.
Variety of incense sticksThere are several brands of incense sticks in the market that release different aromas due to presence of different gases. If consistency is not maintained, results can be affected.To eliminate this error, all incense sticks are procured from the same source – Boswellia serrata.
Number of sapodilla fruits kept for ripeningDifferent amounts of fruits used can affect the resultsIt was ensured that exactly 5 pieces of the fruit was taken for each experiment.
Temperature and humidityDifference in these factors may cause change in resultsAll measurements were taken on the same day. The room temperature was measured as 24°C using a room thermometer and humidity was taken at 60% as printed by the local newspaper
Time of exposure to incense sticksVarying intervals of time can jeopardize the resultsThe time of burning of dhoop and subsequent closing of all boxes was noted using a simple wall clock and calendar. It was ensured that the data was collected at the exactly the same time five days later

Figure 3 - Table On Control Variable

Figure 4 - Table On Apparatus Required

MaterialsQuantitySource
Sapodilla (Manilkara zapota)50 medium sized piecesDirectly from the same farm.
Indian Frankincense (Boswellia serrata)75 gSame brand purchased from store
Distilled water100 ccSchool Laboratory

Figure 5 - Table On Materials Required

Methodology

The main steps for conducting this study were:

  • Procurement of raw sapodilla and incense sticks from the market
  • Carry out ripening in controlled conditions for  a period of 5 days
  • Extract samples from each of the ripened fruits.
  • Measure refractive index of each sample using the refractometer.
  • Comparison of the amount of sugar present in test samples with respect to control samples of sapodilla (where no incense sticks were used to influence the ripening process).
  • Processing of raw data  and its statistical analysis.

Risk Assessment

Apparatus for addressing safety riskUse
Safety gogglesTo prevent contact of chemicals with eyes
GlovesTo prevent contact of chemicals with skin to prevent irritation and rashes
Lab CoatTo provide an additional layer of protection from chemicals and glassware.

Figure 6 - Table On Apparatus For Addressing Safety Risk

Safety ConcernExplanationSolution
Sharp knives are dangerousIt can cause cuts if used improperlyUse carefully only on chopping board, not directly with hand and also in presence of teacher.
Glassware used can get brokenTest tubes, beakers, measuring cylinders, pipettes etc. can be broken accidentally and cause injury to the experimenterPrecaution should be taken while handling glassware. Wear gloves and lab coat Broken glassware should be disposed off separately.
Frankincense sticks can cause smoke and fireThey can cause the cardboard boxes to start burning.They should be mounted on stands while burning such that no glowing part can come in contact with cardboard.

Figure 7 - Table On Safety Concern

Environmental ConcernExplanationSolution
Gases liberated by Frankincense sticks can cause environmental damageIt can cause air pollution, airway disease and health problems.Experiment should be carried out in fume chambers so that the gases are not directly inhaled.

Figure 8 - Table On Environmental Concern

Ethical Concerns

There were no ethical concerns in this project.

Procedure:-

  • Procuring raw sapodilla was done from a farm locally.
  • The Boswellia serrata was prepared in the following manner.
    • Boswellia serrata sticks were crushed in the mortar and pestle
    • A piece of lab paper was  placed onto the weighing scale and then  the scale was tared (reset to zero).
    • Using the spatula, 5.0 g of powdered Boswellia serrata was measured and wrapped in the lab paper itself.
    • Similar packets were created for 10.0 g, 15.0 g, 20.0 g, and 25.0 g of powdered Boswellia serrata and lab paper was wrapped around the Boswellia serrata to secure it.
  • Six boxes were prepared in the following manner:
    • The boxes were numbered from 1-6. The first box did not contain any Boswellia serrata. Boxes 2-6  contained 5.0 g, 10.0 g, 15.0 g, 20.0 g, and 25.0 g of Boswellia serrata respectively.
    • Five  raw fruits were weighed first individually on the mass balance.
    • They were then put into a box, ensuring that they were not touching each other.
    • The measured amount of Boswellia serrata was placed into a steel bowl and the bowl was placed in the box, next to the fruits. This step applied to all boxes except Box 1.
    • Using a matchstick, the Boswellia serrata was lighted on fire and the container’s lid was halfway covered.
    • Once the Boswellia serrata had finished burning, the box was closed and taped shut.
    • Let sit for 5 days.
  • At the end of the said period, the following procedure was followed for contents of Box 1.
    • Each fruit was weighed individually and the final weights noted.
    • Each fruit was then chopped separately and placed in separate bowls.
    • The chopped pieces from one bowl were pureed using a blender.
    • Filter paper was first lined on the funnel and the funnel was placed above a beaker.  The puree was poured on the funnel and the fruit juice (minus the fibres) was collected in the beaker.
    • This was done for all five fruits and the juices were transferred into 5 different test tubes.
    • Using a dropper,  a drop of the juice from one fruit was placed onto the face of the hand refractometer.
    • The measurement was taken and repeated for all five fruits as five trials.
  • The above steps were repeated for  Boxes 2-6.
  • Raw Data was processed and appropriate conclusions were drawn using statistical analysis.

Ouantitative raw data

TrialsInitial Weight ( in g) ±0.1 gFinal Weight (in g) ±0.1 gRefractive index (in %)Observations
157.848.023Wrinkled skin, little white fungus on skin, soft to touch
253.143.718Clear liquid on skin, little wh. ite fungus on skin, soft to touch. Wet patch on box
346.238.417Little clear liquid on skin, soft to touch. Wet patch on box.
443.438.817No visible change (no fungus, no wrinkles, no liquid, firm to touch)
545.739.622Wrinkled skin, little white powdery fungus on skin, mildly soft to touch

Figure 9 - Table On Shows The Initial Weight, Final Weight, Refractive Index Of Sugar Percentage And Visual Observations Of One Manilkara Zapota Before And After Being Placed In A Cardboard Box For 120 Hours.

TrialsInitial Weight ±0.1 gFinal Weight ±0.1 gRefractive index (in %)Observations
151.845.919No visible changes (no fungus, no wrinkles, no liquid, firm to touch)
257.249.517Small patches of fungus near top and on skin, little clear liquid on skin, soft to touch. Wet patch on box.
347.041.115Mildly wrinkled skin, mildly soft to touch, very little fungus,
445.239.416Mildly soft to touch, few fungus patches, especially near top, mildly wrinkled skin.
542.335.516Wrinkles skin, few fungus patches, mostly firm to touch

Figure 10 - Table On Shows The Initial Weight, Final Weight, Refractive Index Of Sugar Percentage And Visual Observations Of One Manilkara Zapota Before And After Being Exposed To The Smoke Of 5 Grams Of Boswellia In A Cardboard Box For 120 Hours.

TrialsInitial Weight ±0.1 gFinal Weight ±0.1 gRefractive index (in %)Observations
148.742.516No visible changes
253.147.016No visible changes
359.450.817No visible changes
442.237.318No visible changes
548.940.420Soft to touch

Figure 11 - Table On Shows The Initial Weight, Final Weight, Refractive Index Of Sugar Percentage And Visual Observations Of One Manilkara Zapota Before And After Being Exposed To The Smoke Of 10 Grams Of Boswellia In A Cardboard Box For 120 Hours.

TrialsInitial Weight ±0.1 gFinal Weight ±0.1 gRefractive index in %Observations
158.849.019Mildly soft to touch, wrinkled skin, white fungal patches
241.435.716Fungus near top, mildly soft to touch, wrinkled skin
353.547.418Mildly wrinkled skin, mildly soft to touch
448.642.917Few fungal patches
547.040.720No visible changes

Figure 12 - Table On Shows The Initial Weight, Final Weight, Refractive Index Of Sugar Percentage And Visual Observations Of One Manilkara Zapota Before And After Being Exposed To The Smoke Of 15 Grams Of Boswellia In A Cardboard Box For 120 Hours.