An Investigation on the Possible Utilization of Red Onion Extract as Blood Detector

Astillo, Rio Roane P. Keh, Kent Edbert T. Ladera, Graciella S. Paz, Gerald John A. Perez, Patria Nia V. Sioson, Berona Faith C.

Outline
Rationale Significance Objectives Methodology Chapter 4 Chapter 5

Rationale, Significance, Objectives

Rationale
We decided on Red onions as our subject of research for it is widely available, very cheap and is practically a household reagent.

Significance of the Study

The red onion farmers Clinical technicians Crime laboratory scientists The community The future researchers

Objectives
Determine the anthocyanin extraction procedure of red onions that is suitable for presumptive blood detection Confirm a color reaction by red onion extract against blood. Determine the color produced by the red onion extract against the following:
Stained fresh blood on wood, white fabric, metal (knife), concrete (floor), and filter paper as wiped, dried blood and fresh blood droplets

Objectives
Stained diluted blood on wood, white fabric, metal (knife), concrete (floor), and filter paper as wiped, dried blood and diluted blood droplets Red liquid (represents fake blood) on wood, white fabric, metal (knife), concrete (floor), and filter paper

Methodology

Methodology
Research design Qualitative research The study covers the different anthocyanin extraction methods.

Extraction Method A The samples were soaked in 200mL solution (w/v, 1:2) of ethanol for 2448 hours at room temperature. The mixture was filtered on a Buchner funnel and the flask and the plant material were rinsed with fresh portions of alcohol. The filtrate was concentrated using the hot plate apparatus at 80C. The solution obtained was tested with H2O2

Extraction Method B. The samples were soaked in 200mL solution (w/v, 1:2) of ethanol for 24-48 hours at room temperature. The mixture was filtered on a Buchner funnel and the flask and the plant material were rinsed with fresh portions of alcohol. The filtrate was concentrated using the soxhlet apparatus at 78C. The solution obtained was divided into two.
w/ H2O2 w/o H2O2

Extraction Method C The samples were soaked in 200mL solution (w/v, 1:2) of ethanol for 24-48 hours at room temperature. The mixture was filtered on a Buchner funnel and the flask and the plant material were rinsed with fresh portions of alcohol. The filtrate was concentrated using the soxhlet apparatus at 78C and hot plate apparatus at 80C. The solution obtained was divided into two.
w/ H2O2 w/o H2O2

Extraction Method D. The samples were soaked in 200mL solution (w/v, 1:2) of distilled water for 24-48 hours at room temperature in darkness. The mixture was filtered on a Buchner funnel and the flask and the plant material were rinsed with fresh portions of distilled water. The solution obtained was divided into two:
w/ H2O2 w/o H2O2

Extraction Method E The samples were soaked in 200mL solution (w/v, 1:2) of methanol with 4% citric acid for 2-3 hours at room temperature in darkness. The mixture was filtered on a Buchner funnel and the remaining solids were washed with 4.0% citric acid in methanol until a clear solution was obtained. The combined filtrates were concentrated using rotary evaporator at 60C. The extract was dissolved in a solution (v/v, 1:1) of 0.4% citric acid (w/v) in distilled water and the solution obtained was used as a blood detector.

II. Preparation of different samples A. Stained blood Blood samples were obtained Divided into two separate red top tubes
first tube was maintained fresh while second tube was subject to dilution with the ratio of 1:20.

Cell lysis is expected for better reaction. B. Red Liquid Red crepe paper was cut into small pieces and soaked in tap water until it produced intense red color similar to blood.

III. Test for detection of blood The fresh, and diluted blood, and red liquid was applied to the different surfaces. Each surface was tested with 3 trials with equal amount of blood.

3.1
3.2

Wood 3.1.1 Dried blood 3.1.2 Blood droplets White Fabric 3.2.1 Dried blood 3.2.2 Blood droplets

3.3
3.4 3.5

Metal (knife) 3.3.1 Dried blood 3.3.2 Blood droplets Concrete (floor) 3.4.1 Dried blood 3.4.2 Blood droplets Filter paper 3.5.1 Dried blood 3.5.2 Blood droplets

Extract 2-3 drops/ sample Positive: Green Control: Guaiacs Reagent 2-3 drops/ Sample Positive: Blue

Reading time: After 5 minutes

Research Flow Chart

Select Topic

Gather Information

Research Methodology

Collection of Onion bulb and peels

A. Ethanol Hotplate Extraction (80C) B. Ethanol Soxhlet Extraction (78C) Extraction Method

w/H2O2 w/H2O2

w/o H2O2
w/H2O2

C. Ethanol Soxhlet-Hotplate Extraction (78C)

w/o H2O2
w/H2O2

Prep. Of Samples

D. Distilled Water (Aqueous) Extraction E. Acidified Methanol Rotary Evaporator (60C)

w/o H2O2
w/o H2O2

Preparation of Samples Experimentation and Testing of Samples

Reading of Samples

Interpretation

Summary and Conclusion

Recommendation

Chapter IV
Presentation and Interpretation of Data

Method A
Method B Method C Method D

:
: : :

Method E

Ethanolic extraction using hot plate at 80 C Ethanolic Extraction using Soxhlet at 78 C Ethanolic Extraction using Soxhlet at 78 C and hot plate at 80C Aqueous Extract of red onion Rotary Evaporation at 60C of 4% Citric Acid in Methanol

Guide Pictures for color reaction

(+) Diluted Blood Dry (+) (-)

Diluted Blood Droplet

Fresh Blood Dry

Fresh Blood Droplet

Summary of reactions of different extraction methods

Method A Method B Method C Method D Method E Control: Guaiacs Reagent Negative Negative Negative Negative Positive Positive ( with some negatives)

Chapter IV

Chapter V
Summary, Conclusion, and Recommendation

Method 1: Hot Plate of Ethanolic Extract at 80 C

All 12 trials on the surface of wood, white cloth, knife, filter paper and concrete material resulted negative. The red liquid acquired all yielded negative results; no green color was observed in all trials. All trials on Guiacs reagent resulted positive; blue color was observed in all trials

Method 2: Ethanolic Extraction using Soxhlet at 78 C

All 12 trials (with or without hydrogen peroxide) on the surface of wood, white cloth, knife, filter paper and concrete material resulted negative. The red liquid acquired all yielded negative results; no green color was observed in all trials. All trials on Guiacs reagent resulted positive; blue color was observed in all trials

Method 3: Ethanolic Extraction using Soxhlet at 75 C and hot plate at 80C

All 12 trials (with or without hydrogen peroxide) on the surface of wood, white cloth, knife, filter paper and concrete material resulted negative. The red liquid acquired all yielded negative results; no green color was observed in all trials. All trials on Guiacs reagent resulted positive; blue color was observed in all trials

Method 4: Aqueous Extract of red onion

All 12 trials (with or without) on the surface of wood, white cloth, knife, filter paper and concrete material resulted negative. The red liquid acquired all yielded negative results; no green color was observed in all trials. All trials on Guiacs reagent resulted positive; blue color was observed in all trials

Method 5: Rotary Evaporation at 60C of 4% Citric Acid in Methanol

All 12 trials on the surface of white cloth and filter paper resulted positive. On the wood surface, there were 11 positive results and 1 weak positive result obtained. However, on the metal surface, only 10 positive results were obtained and 2 weak positive results.

Method 5: Rotary Evaporation at 60C of 4% Citric Acid in Methanol

Lastly 4 positive and 8 weak positive results were detected on the concrete surface. The red liquid acquired all yielded negative results; no green color was observed in any trials.

Guiacs test
The Guiacs test on wood produced 12 positive results. However, on the surface of the white cloth and filter paper, there were 11 positive results and 1 weak positive result obtained on each surfaces. Lastly, on the surface of metal and concrete material, 11 resulted positive and 1 resulted negative on each surfaces.

Conclusion
The researchers conclude that red onion extract using methanol with 4% Citric acid can be used best on paper and cloth surfaces followed by wood and metal surfaces in the presence of blood.

Recommendation
To try blood detection on other surfaces To try other extraction method to extract anthocyanin To try other sources of anthocyanin To device method to extract pure anthocyanin To use different concentration of the pure anthocyanin extract

RRL