Organic Chemistry and Functional Group Identification in Butterfly Pea (Clitoria ternatea)

Pichayathida Siriwechdaruk

Butterfly pea
Butterfly pea (Clitoria ternatea) is a climbing plant whose flower is blue and is
commonly used as natural food coloring. The origin of butterfly pea is believed to be the South
America and Asia. It is commonly cultivated for its azure flower with winged petals, which last
for 24 hours only. It has many uses as it is commonly used in culinary products as dyes, and is
used as ayurvedic medicine in India [10] . The genus name of the plant is from the fact that the
shape of the flower resembles the shape of female genital organ and the name of the species,
“Ternatea” comes from the name of the Eastern Indonesian island, Ternate [15] . All parts of the
plant are being used for medicinal purposes as it was proven to have the cooling, anti-
inflammatory, antifungal, antioxidant, stimulative, laxative and diuretic properties. These
benefits from ingesting the plant was from the fact that many chemicals could be extracted from
it such as: homopterocarpin, anthocyanin $% , delphinidin $& , pentanal, cyanoacetic acid,
hexadecanoic acid, isoamyl acetate, and p-hydroxycinnamic acid [11] , in which some of the
chemicals carries beneficial properties to human health.

Cyanoacetic acid

Cyanoacetic acid ('3 (3 N)2 ) is a white to yellow solid with an unpleasant odor. It has the
melting point of 65 Celsius and boiling point of 108 Celsius. It is soluble in water and has the
*+, of 2.45 at standard state [3] . Cyanoacetic acid could be found in methanolic extract of
butterfly pea flowers [1] . Cyanoacetic is usually used in aiding chemical reactions such as
esterification of cyanoacetic acid (chloride) and t-butanol to obtain t-butyl cyanoacetate. It also
could be used as a substitution for an a-ketoacytyl anion in the diastereoselective synthesis of a
decalin fragment of the natural product insecticide azadirachtin [7] .
Delphinidin

Delphinidin, ('15 (11 )7- ) is an anthocyanin component that could be easily found in
berries and red wine. It is also commonly found in blue flowers; hence, it could be found in
butterfly pea flowers. Delphinidin, along with other anthocyanins, is known as the principle and
basic components of flower color pigments [6] . It holds the molecular weight of 303.246 g/mol,
commonly found in the form of powder. It is water soluble [4] and like some other anthocyanins,
the color of delphinidin is pH dependent; hence, it could also be used as pH indicator. When
delphinidin is treated with acid, the color would shift to be more red and when it is treated with
basic solution, it will appear more blue. At neutral state, delphinidin inhabits the color of blue-ish
purple [12] . Other then its use as a pH indicator, delphinidin is highly proven that it contains the
properties of being an antioxidant [6] , having anti-inflammatory properties and having the
property of being a potent angiogenic inhibitor [8] .

Functional group and identification

Cyanoacetic acid contains two functional groups: Carboxylic acid and nitrile group. The
test for carboxylic acid could be easily carried out by adding baking soda or sodium hydrogen
carbonate (./(')3 )to the solution. If carbon dioxide bubble were formed due to the reaction,
then the test is positive and the compound does contain carboxyl group.
1'))( + ./(')3 − − − − − − − − − −> 1'))./ + (2 ) + ')2 (6)
For nitrile group, it could be identified by adding 1 cc of the solution and 20 cc of absolute
alcohol to a round bottom flask. Then, by using a reflux condenser, immerse the flask, until the
neck, in a 50-60 Celsius water bath, following by gradually adding 1.5 grams of sodium to the
solution through the top of the condenser. After 10 to 15 minutes, all of the sodium should have
reacted with the solution and the reduction should be complete. Let the solution cool down and
add 10 cc of concentrated hydrochloric acid to the mix. Following the reduction, set up a system
for distillation, distilling the alcohol from the mix. After the distillation is finished, add a solution
prepared from 6 grams of sodium hydroxide in 6 cc of water. Then, reconnect the flask to the
condenser again and use a smoky flame until all of the contents are almost dry, catching the
distillate in a flask containing 3 cc of water then test for the pH of the end product. If the end
product (the distillate) is alkaline, then the compound contains nitrile group [2] This is because
the nitrile group had reacted with the added chemicals and gives out an distillate which is
alkaline.
For delphinidin, there are four functional groups: alcohol, resorcinol, pyrogallol, and
pyrium salt. The hydroxyl group (alcohol) could be easily identified with the use of the ceric
ammonium test. If the solution reacts with ceric ammonium nitrate and form a red color
compound, alkoxy cerium (IV), then the test is positive and the solution contains alcohol [7] .
21 − )( + (.(4 )2 '9(.)3 )4 − − − − − − − − − −> (1)()2 '9(.)3 )4 + 2.(4 .)3
Yellow Pink/Red
For resorcinol, it could be identified through “fluorescein reaction”. When Phthalic anhydride is
added with resorcinol and treated with sulfuric acid, it will gives a dye fluorescein. If the mixture
was treated with sodium hydroxide solution and gives yellow-ish-red solution with green
fluorescence, then the test is positive and the solution contains resorcinol [5] .
Product: Materials & Apparatus
1. Butterfly pea flower (Clitoria ternatea, sun dried)
2. Kaffir (Citrus hystrix)
3. Ginger (Zingiber officinale)
4. Sodium Chloride (./':)
5. Sodium Laureth Sulfate ('24 (50 ./2 )5 ;)
6. Sodium Lauryl Sulfate ('12 (25 ./)4 ;)
7. Baking soda (./(')3 )
8. Pot with lid
9. Cutting board
10. Knife
11. Measuring spoons
12. Electronic balance
13. Strainer
14. Graduated jug
15. Bottle
Product: Method
1. Clean the kaffir and ginger thoroughly with water, make sure that there is no soil left on
the ginger
2. Slice 450 g of kaffir and 57g of ginger into thin slices, put them into the pot
3. Wash 10g of dried butterfly pea flowers thoroughly with water, making sure to remove
the anther and carpel as the pollens and the parts could cause irritation of the scalp.
4. Drain the washed butterfly pea flowers by using a strainer and put the flowers into the pot
5. Add 700 mL of water into the pot along with the herbs. Simmer with low heat for 1.5
hours or until all of the herbs are soft and mushy. Make sure to keep the lid on all the
time to reduce loss of aromatic oil.
6. Drain the mixture with strainer, keeping the stock
7. Add 2 tablespoon of baking soda in order to make the solution less acidic. Stir until all of
the baking soda dissolves
8. Measure 400 mL of the stock obtained in part 5 to 100 mL of Sodium Laureth Sulfate,
stir until they are combined
9. Add 1 teaspoon of sodium lauryl sulfate to the mix. Stir until they are combined
10. Add 7 teaspoons of sodium chloride to the mix. Stir until they are combined. If the
shampoo mix is not as thick as wanted, then gradually add more sodium chloride to the
mix until meet the desired consistency.
11. Pour the shampoo into a bottle to store
Chemistry behind the product
When the butterfly pea flower was put into water, the color of the water changed to be
appearing as slightly blue as the anthocyanins from the flower were mixed with the water at
neutral pH. As the butterfly pea flowers, kaffir, and ginger was being boiled in water, chemicals
such as delphinidin, other anthocyanins that the flower contains, phenolics, flavonoids from
butterfly pea was released along with other chemicals from kaffir and ginger such as essential
oil, kaffir lime juice, polyphenols, tannin, and flavonoid from ginger. Due to the acidity of the
kaffir juice, the color of the anthocyanins, including delphinidin, was changed due to the low pH
(pH~3.5) of the solution, making the solution appears reddish-violet. However, when the
solution finished boiling and was left to cool, baking soda, which is alkaline, was added to the
solution to lessen the acidity and the pH was increased to be only slightly acidic (pH~5.5);
hence, the color of the solution has been changed back to blue-ish purple once again. When the
baking soda was added to the solution, bubbles were formed as the baking soda (sodium
hydrogen carbonate) reacts with the acid, resulting carbon dioxide bubbles to form.
Sodium lauryl sulfate and sodium laureth sulfate, which are surfactants, were later added
to the solution. The two chemicals will give the shampoo the ability to clean off the excess oil
and dirts as it has ability to lower surface tension, break up molecules on the surface layer as it
has both water soluble and oil soluble parts [13] . After the stock and the two surfactants were
combined, sodium chloride was added in order to make the shampoo less runny. The viscosity of
the solution was increased due to the increase of the sodium chloride concentration through the
increase of ionic strength of the solution [9] .
 References
[1] Al-Snafi, A. E. (2016). Pharmacological importance of Clitoria ternatea - A review. IOSR
Journal of Pharmacy,6(3). Retrieved from
http://www.iosrphr.org/papers/v6i3/G0636883.pdf
[2] Cutter, H. & Taras, M. (1941). A Method for Identification of Nitriles. Industrial and
Engineering Chemistry.13(11). Retrieved from
https://pubs.acs.org/doi/abs/10.1021/i560099a031
[3] Cyanoacetic acid (n.d.). Retrieved 20 April, 2018 from
http://www.chemicalbook.com/ChemicalProductProperty_EN_CB8217349.htm
[4] Delphinidin Cation. (2005). Retrieved 20 April, 2018 from
https://pubchem.ncbi.nlm.nih.gov/compound/Delphinidin#section=Top
[5] Detection of Functional Group. (n.d.) Retrieved 20 April, 2018 from
http://vlab.amrita.edu/?sub=2&brch=191&sim=345&cnt=1
[6] Estevez, L & Mosquera, R.A. (2008). Molecular Structure and Antioxidant Properties of
Delphinidin. Retrieved 18 April, 2018 from
https://pubs.acs.org/doi/abs/10.1021/jp8043237
[7] Freeman, F. (n.d.). Cyanoacetic acid. Retrieved 18 April, 2018 from
http://reag.paperplane.io/00000774.htm
[8] Lamy, S., Blanchette, M., Michaud-Lavesque, J., Lafleur, R., Durocher, Y., Moghrabi, A.,
Barrette, S., Gingras, D. & Beliveau, R. (2006). Delphinidin, a dietary anthocyanin,
inhibits vascular endothelial growth factor receptor-2 phosphorylation.
Carcinogenesis,27(5). Retreived from
https://academic.oup.com/carcin/article/27/5/989/2476121
[9] Ozbek, H., Fair, J.A., & Phillips, S.L. (2010). Viscosity of aqueous sodium chloride solutions
from 0-150 C. Retrieved 20 April, 2018 from
https://cloudfront.escholarship.org/dist/prd/content/qt3jp6n2bf/qt3jp6n2bf.pdf
[10] Pwee, T. (2016). Butterfly Pea. Retrieved 22 March, 2018 from
http://eresources.nlb.gov.sg/infopedia/articles/SIP_763_2004-12-20.html
[11] Neda, G. D., Rabeta, M. S. & Ong, M. T. (2013). Chemical composition and
anti-proliferative properties of flowers of Clitoria ternatea. International Food Research
Journal,20. Retrieved from http://www.ifrj.upm.edu.my/20%20(03)%202013/28%20I
FRJ%2020%20(03)%202013%20Rabeta%20(389).pdf
[12] Saptarini,N. M., Suryasaputra, D. & Nurmalia, H. (2015). Application of Butterfly Pea
(Clitoria ternatea Linn) extract as an indicator of acid-base indicator. Journal of
Chemical and Pharmaceutical Research,7. Retrieved from
http://www.jocpr.com/articles/application-of-butterfly-pea-clitoria-ternatea-linn-extract-a
s-an-indicator-ofacidbase-titration.pdf
[13] Schaefer, A .(2016). What is Sodium Laurly Sulfate?. Retrieved 20 April, 2018 from
https://www.healthline.com/health/beauty-skin-care/what-is-sodium-lauryl-sulfate
`[14] Suebkhampet, A. & Sotthibandhu, P. (2012). Effect of Using Aqueous Crude Extract From
Butterfly Pea Flowers (Clitoria ternatea L.) as a Dye on Animal Blood Smear Staining.
Suranaree J. Sci. Technol.19. Retrieved from http://www.thaiscience.info/Journals/Ar
ticle/SJST/10890421.pdf
[15] Upmanyu, N., Chauhan, N.S., Tripathi, A., Mishra, P., Shah, K., Gupta, J.K., Singh, N.K.
(2017). A Review on Clitoria ternatea(Linn): Chemical and Pharmacology. In Medicinal
Plants and its Therapeutic Uses (7). Retrieved from
https://www.esciencecentral.org/ebooks/ebookchapter/a-review-on-clitoria-ternatea-linn-
chemistry-and-pharmacology-434/7#4-tabreflink