ArticlePDF Available

Phytochemical screening of Broussonetia luzonicus (Moraceae) leaves

Authors:

Abstract and Figures

This is concerned with the identification of the phytochemical that is contained in the leaves of the tree Broussonetia luzonicus the leaves of which are commonly eaten in the northern regions of the Philippines. Other studies of other Broussonetia spp. have been reported to have antibacterial, antioxidant, antitumor, and pancreatic lipase inhibitory activity. This study sought to identify the phytochemical that the Broussonetia luzonicus leaves contain. The study began with the collection and identification of B. luzonicus leaves. The crude methanolic extract was collected by percolation and concentrated using a rotary evaporator. It was then tested using qualitative phytochemical screening methods and it yielded carbohydrates, reducing sugars, flavonoids, phenolic compounds, alkaloids, and sterols. © 2016, Journal of Chemical and Pharmaceutical Research. All rights reserved.
Content may be subject to copyright.
Available online www.jocpr.com
Journal of Chemical and Pharmaceutical Research, 2016, 8(2):335-338
Research Article
ISSN : 0975-7384
CODEN(USA) : JCPRC5
335
Phytochemical screening of Broussonetia luzonicus (Moraceae) leaves
Choa John Benson D.
1
, Lu Roanne V.
2
, Nombrado Mark A.
1
, Rayos Garina Kaye R.
1
,
Invento Chelsea Dae
1
and Castañeda Gerald
1
1
College of Pharmacy, National University- Sampaloc, Metro Manila, Philippines
2
Lyceum of the Philippines University- Governor’s Drive, General Trias, Cavite, Philippines
_____________________________________________________________________________________________
ABSTRACT
This is concerned with the identification of the phytochemical that is contained in the leaves of the tree Broussonetia
luzonicus the leaves of which are commonly eaten in the northern regions of the Philippines. Other studies of other
Broussonetia spp. have been reported to have antibacterial, antioxidant, antitumor, and pancreatic lipase inhibitory
activity. This study sought to identify the phytochemical that the Broussonetia luzonicus leaves contain. The study
began with the collection and identification of B. luzonicus leaves. The crude methanolic extract was collected by
percolation and concentrated using a rotary evaporator. It was then tested using qualitative phytochemical
screening methods and it yielded carbohydrates, reducing sugars, flavonoids, phenolic compounds, alkaloids, and
sterols.
Keywords: Broussonetia luzonicus, Alukon, himbabao phytochemical screening, methanolic extract, Moraceae
_____________________________________________________________________________________________
INTRODUCTION
Plants have been important sources of drug products since ancient times. These plants contain secondary metabolites
which are necessary for the survival of the species, and several of these compounds such as digoxin, colchicine, and
paclitaxel have been utilized as drugs for men and animals. Broussonetia luzonicusis a tree which is endemic in the
Philippines. It belongs to the family Moraceae. B. luzonicus is commonly known as Alukon in Ilocano (a major
Philippine language spoken in the northern part of the Philippines) and as Himbabaoin Tagalog (a major Philippine
language spoken in Manila and its surrounding areas) The tree can grow up to 10 meters and 40 cm in diameter and
the leaves are simple, alternate, ovately oblong, membranous, 15 cm in length and 7 cm in width, acute or acuminate
[1]
. The leaves are commonly eaten as vegetable by the residents of the Ilocos region of the Philippines. According to
previous studies of Broussonetia spp., B. kazinokihas been known to have tyrosinase inhibitory activity which can
reduce hyperpigmentation in hypertrophic scars
[2]
. While B. papyrifera phenolic compounds have been reported to
have estrogen synthesis-inhibiting and antioxidantactivity
[3]
. It was also reported that B. papyrifera flavonoids
showed antimicrobial properties
[4]
.And that B. kazinoki alkaloids can inhibit α-glucosidase activity
[5]
. Presently,
there has been only one study ofB. luzonicus, that undertaken by Ragasa
[6]
which identified the constituents of the
DCM extract of B. luzonicus leaves. The study indicated the presence of lupenone, squalene, β-carotene, vitamin K,
β-sitosterol, and epitaraxerol.
Statement of the Problem
The problem that this researcher sought to answer was the following: What are the phytochemical of constituents of
Broussonetia luzonicus leaves.
Choa John Benson D. et al J. Chem. Pharm. Res., 2016, 8(2):335-338
______________________________________________________________________________
336
This present study sought to determine the constituents of the crude methanolic of Broussonetia luzonicus leaves
using qualitative phytochemical screening methods to assess the potential of the plant for drug discovery. The results
of the result of the experiment indicate of the presence of carbohydrates, reducing sugars, flavonoids, phenolic
compounds, alkaloids, and sterols.
EXPERIMENTAL SECTION
Collection and Authentication of Broussonetia luzonicus leaves
One thousand and three hundred grams (1300g) of fresh leaves was collected at Santa Fe, Nueva Vizcaya. The plant
specimen was authenticated by Manuel D. Ching, a botanist from the Bureau of Plant Industry (BPI). The specimen
was deposited with the document number (PLT-ID-CRD-256-15) as certification for plant authentication.
Extraction of the crude methanolic extract of Broussonetia luzonicus leaves
The leaves were air-dried and ground using a blender. The ground leaves were extracted by percolation using
methanol as the solvent. Exhaustive extraction was used to obtain more extracts from the leaves. After collecting the
extracts, it was concentrated under reduced pressure using a rotary evaporator.
Phytochemical screening of the crude methanolic extract of Broussonetia luzonicus leaves
The phytochemical screening was done at the Institute of Pharmaceutical Sciences, National Institutes of Health
using the following qualitative methods: Molisch test for carbohydrates, Fehling’s test for reducing sugars, alkaline
reagent test, lead acetate test, and magnesium hydrochloride test for flavonoids, ferric chloride test and gelatin test
for tannins, Borntrager’s test for anthraquinones, Keller-Kiliani’s test for cardiac glycosides, Wagner’s test, Mayer’s
test, and Hager’s test for alkaloids, Liebermann-Buchard’s test and Salkowski’s test for sterols and terpenoids, froth
test for saponins, and acetone-water test for risins.
RESULTS AND DISCUSSION
Percentage yield of the crude methanolic extract of Broussonetia luzonicus leaves
The percentage yield was computed using the formula:
ℎℎ()
ℎℎ() 100
The amount of extract obtained from 1300 grams of dried leaves was 273.663 grams and it yielded 21.048%.
Phytochemical screening of the crude methanolic extract of Broussonetia luzonicus leaves
The phytochemical screening produced the following result:
As seen in Table 1, the tests are indicative of the presence of carbohydrates, reducing sugars, flavonoids, tannins,
alkaloids, and sterols. Phytochemicals such as flavonoids which are polyphenolic compounds have been known to
have powerful antioxidant activity that can reduce risk of coronary diseases, it can also exhibit a wide range of
activity such as anti-inflammatory, antiviral, antibacterial, antiulcer, antiosteoporotic, antiallergic, and antihepatic
action
[7]
. Tannins are also polyphenolic compounds which is also known to possess powerful antioxidant activity
which are used against heart disease through reducing lipid oxidation
[8]
. Alkaloids are basic nitrogenous compounds
which are pharmacologically-active, and which may exhibit tranquilizing and stimulating activity on the nervous
system, hypertensive and hypotensive action, vasoconstrictor and vasodilator effect on the cardiac system, Alkaloids
they can also affect the transmitter actions on the muscular system
[9]
. Phytosterols or plant sterols have a chemical
structure similar to cholesterols which have been reported to decrease cholesterol absorption and plasma Low
Density Lipoprotein (LDL) values
[10]
. These results suggest that Broussonetia luzonicus may be a potential
candidate to be further developed in to a drug compound.
Choa John Benson D. et al J. Chem. Pharm. Res., 2016, 8(2):335-338
______________________________________________________________________________
337
Table 1: Phytochemical Analysis of Broussonetia luzonicus leaves
Phytochemical Analysis of Broussonetia luzonicus leaves
Phytochemical Name of Test Theoretical Result Actual Result Indication
Carbohydrates Molisch Test Violet ring at the junction Formation of violet ring at the
junction
(+)
Reducing Sugars Fehling’s Test Formation of brick red precipitate Formation of brick red
precipitate (+)
Flavonoids
Alkaline Reagent Test Yellow coloration which disappears upon
the addition of dilute acid
With alkaline reagent:
Yellowish green color persisted
With dilute acid: Discharge of
yellow tinge in the solution
(+)
Lead Acetate Test Formation of yellow colored precipitate Formation of yellowish green
colored precipitate (+)
Magnesium
Hydrochloride
Reduction Test
Red or orange coloration of the solution Appearance of a green colored
solution (-)
Tannins Ferric Chloride Test Blue or green to black coloration of the
solution Formation of a green colored
solution (+)
Gelatin Test Formation of white precipitate Formation of a turbid yellowish
green solution
(-)
Glycosides
Borntrager’s Test
(Anthraquinone
Glycoside)
Pink, red or violet coloration in the
ammoniacal layer Appearance of a pale yellow
solution (-)
Keller Killiani’s Test
(Cardiac Glycoside) Appearance of reddish brown or purple ring
at the junction Appearance of a green to brown
ring at the junction (-)
Alkaloids Wagner’s Test Formation of reddish brown precipitate or
turbidity
Formation of turbid solution (+)
Mayer’s Test
Formation of turbid solution
(+)
Hager’s Test Formation of yellow precipitate or turbidity Formation of turbid solution (+)
Steroids and
Terpenoids
Liebermann-Burchard’s
Test
Formation of reddish brown ring / deep red
(triterpenoid) or green (sterol) coloration on
the upper layer of the solution
Appearance of a brown ring at
the junction with a green color
on the upper layer (+)
Salkowski’s Test Red (sterol) or yellow (triterpenoid)
coloration in the lower layer of the solution No change in the color of the
original solution (-)
Saponins Froth Test Formation of honey-comb froth greater than
2 cm from the surface of the extract
No formation of honey-comb
froth
(-)
Resins Acetone – Water Test Solution becomes turbid Appearance of a clear yellow
solution (-)
+ = Present; - = Absent
CONCLUSION
In this study, the phytochemical screening of Broussonetia luzonicus indicated the presence of carbohydrates,
reducing sugars, flavonoids, tannins, alkaloids, and sterols. These secondary metabolites could be a source of
potential drug compounds which can be used against various diseases such as cardiovascular diseases, cancer,
obesity, diabetes, and infections. The results of the study confirm the finding of the study of Ragasa. This study
suggests that Broussonetia luzoniscus contains active compounds which are interesting for further pharmaceutical
research. It is therefore, recommended that further research be conducted in the pharmacologic activity,
characterization, and isolation as well as toxicology of the constituents of Broussonetia luzonicus.
Acknowledgment
This study would like to thank the National University College of Pharmacy and the National Institutes of Health-
Institute of Pharmaceutical Science UP Manila for their support. The authors would also like to acknowledge the
editorial assistance of the Research and Innovation Center of LPU-Cavite.
REFERENCES
[1] Natural Resources Management Center (Philippines) (1986). Guide to Philippine Flora and Fauna. Natural
Resources Management Center, Ministry of Natural Resources and University of the Philippines; Manila: Goodwill
Bookstore.
[2] Baek, Y.S., Ryu, Y.B., Curtis-Long, M.J. Ha., T.J., Rengasamy, R., Yang, M.S. et. al. (2009). Bioorganic and
Medicinal Chemistry, 17(1):35-41. Doi: 10/1016/j.bmc.2008.11.022.
Choa John Benson D. et al J. Chem. Pharm. Res., 2016, 8(2):335-338
______________________________________________________________________________
338
[3] Yang, C., Li, F., Du, B., Chen, B., Wang, F., & Wang, M. (2014). Isolation and Characterization of New
Phenolic Compounds with Estrogen Biosynthesis-Inhibiting and Antioxidation Activities from
Broussonetiapapyrifera Leaves. PLoS One, 9(4).
[4] Sohn, H.-Y., Son, K.H., Kwon, C.-S., Kwon, G.-S. Kwon, & Kang, S.S. (2004). Phytomedicine, 11, 666-672.
[5] Shibano, M., Satoshi, K., Satoko, N., Akazawa., N., &Kusano, G. (1997). Chemical and Pharmaceutical
Bulletin, 45(4): 700-705.
[6] Ragasa, C.Y., Tsai, P.-w., De Castro-Cruz, Kathlia A., and Shen C.-C. (2012). Pharmacognosy Journal. 4(31).
doi: 10.5530/pj.2012.31.1.
[7] Amić, D., Davidović-Amić, D., Bešlo, D., Rastija, V., Lučić, B., & Trinajstić, N. (2007). Current Medicinal
Chemistry, 14, 827-845.
[8] Zhang, L.-l. & Lin, Y.-m. (2008). Journal of Zhejiang University Science B, 9(6): 407-415.
[9] Nestrova, Y.V., Povetieva, T.N., Suslov, N.I., Semenov, A.A., &Pushkarskiy, S.V. (2011). Bulletin of
Experimental Biology and Medicine, 151(4): 425-428.
[10] Lin, X., Racette, S.B., Lefevre, M., Spearie, C.A., Most, M., Ma, L. et al. (2010). European Journal of Clinical
Nutrition, 64, 1481-1487.
ResearchGate has not been able to resolve any citations for this publication.
Article
Full-text available
Broussonetia papyrifera leaves (BPL) as a traditional Chinese medicine are also used in livestock feed for stimulating reproduction, adipose tissue and muscle development; however, the mechanism of their action is still unknown. Through estrogen biosynthesis-guided fractionation in human ovarian granulosa-like KGN cells, five new phenolic glycosides, broussoside A-E(1-5), along with fifteen known dietary phenolic compounds, were isolated from the n-butanol extract of BPL, and their structures were elucidated on the basis of NMR spectra analysis and chemical evidence. New compounds 3, 4, 5 and the known compounds 9 and 10 were found to potently inhibit estrogen biosynthesis in KGN cells. In addition, compounds 9, 17, 18, and 20 showed strong antioxidant activity against ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) and DPPH (1, 1'-diphenyl -2-picryl-hydrazyl radical) assays. These findings suggest that BPL may improve meat quality through the regulation of estrogen biosynthesis. Furthermore, they may be useful for the discovery of potential aromatase modulators from natural products. Finally, they could be considered as a new source for natural antioxidants.
Article
Full-text available
Cited By (since 1996):83, Export Date: 23 March 2014, Source: Scopus
Article
Full-text available
The leaves of Broussonetia luzonicus afforded epitaraxerol (1), lupenone (2), squalene (3), β-carotene (4), vitamin K (5) and β-sitosterol (6), while the flowers yielded 2, 6, lupeol (7), betulin aldehyde fatty acid ester (8) and lupeol fatty acid ester (9). The compounds were isolated by silica gel chromatography and identified by NMR spectroscopy. Triterpenes 1, 2 and 8 were tested for cytotoxicity using the MTT assay. They did not exhibit cytotoxic effect against a human cancer cell line colon carcinoma (HCT 116). Results of the antimicrobial tests on 1, 2 and 8 indicated moderate antifungal activity against C. albicans and low antimicrobial activity against T. mentagrophytes, A. niger, S. aureus, E. coli, P. aeruginosa, and B. subtilis.
Article
Antimicrobial activity of the 18 prenylated flavonoids, which were purified from five different medicinal plants, was evaluated by determination of MIC using the broth microdilution methods against four bacterial and two fungal microorganisms (Candida albicans, Saccaromyces cerevisiae, Escherichia coli, Salmonella typhimurium, Staphylococcus epidermis and S. aureus). Papyriflavonol A, kuraridin, sophoraflavanone D and sophoraisoflavanone A exhibited a good antifungal activity with strong antibacterial activity. Kuwanon C, mulberrofuran G, albanol B, kenusanone A and sophoraflavanone G showed strong antibacterial activity with 5-30 microg/ml of MICs. Morusin, sanggenon B and D, kazinol B, kurarinone, kenusanone C and isosophoranone were effective to only gram positive bacteria, and broussochalcone A was effective to C. albicans. IC50 values of papyriflavonol A, kuraridin, sophoraflavanone D, sophoraisoflavanone A and broussochalcone A in HepG2 cells were 20.9, 37.8, 39.1, 22.1, and 22.0 microg/ml, respectively. These results support the use of prenylated flavonoids in Asian traditional medicine to treat microbial infection and indicate a high potential for prenylated flavonoids as antimicrobial agents as well as anti-inflammatory agents.
Guide to Philippine Flora and Fauna. Natural Resources Management Center, Ministry of Natural Resources and University of the Philippines
Natural Resources Management Center (Philippines) (1986). Guide to Philippine Flora and Fauna. Natural Resources Management Center, Ministry of Natural Resources and University of the Philippines; Manila: Goodwill Bookstore.
Bioorganic and Medicinal Chemistry
  • Y S Baek
  • Y B Ryu
  • M J Curtis-Long
  • T J Ha
  • R Rengasamy
  • M S Yang
Baek, Y.S., Ryu, Y.B., Curtis-Long, M.J. Ha., T.J., Rengasamy, R., Yang, M.S. et. al. (2009). Bioorganic and Medicinal Chemistry, 17(1):35-41. Doi: 10/1016/j.bmc.2008.11.022.
  • L Zhang
  • Y Lin
Zhang, L.-l. & Lin, Y.-m. (2008). Journal of Zhejiang University Science B, 9(6): 407-415.
  • X Lin
  • S B Racette
  • M Lefevre
  • C A Spearie
  • M Most
  • L Ma
Lin, X., Racette, S.B., Lefevre, M., Spearie, C.A., Most, M., Ma, L. et al. (2010). European Journal of Clinical Nutrition, 64, 1481-1487.
  • C Y Ragasa
  • P.-W Tsai
  • De Castro-Cruz
  • A Kathlia
  • C.-C Shen
Ragasa, C.Y., Tsai, P.-w., De Castro-Cruz, Kathlia A., and Shen C.-C. (2012). Pharmacognosy Journal. 4(31). doi: 10.5530/pj.2012.31.1.