CHEMICAL COMPOSITION OF SPIRULINA PLATENSIS OF NOMAYOS-YAOUNDE (CAMEROON)

Abstract

ABSTRACT
Background: S. platenis is rich in macronutrients and micronutrients. It is used as a dietary supplement and found in tropical and semi-tropical areas This study aims to investigate the chemical composition of S. platensis of Nomayos (Yaounde, Cameroon) focusing on macro and micronutrients and phyotchemical bioactive molecules Materiel and methods: S. platensis was collected in a farm in Nomayos (Yaounde, Cameroon). The aqueous extract was used for the determination of nutrients using standard methods. Trace elements were determined by atomic absorption spectrophotometry. The phytochemical composition was determined by using HPLC. Phycocyanin and Carotenoids were determined using chemicals methods.
Results: The extraction yield obatined was 16.84%. The results showed that S platensis contains protein (375.5+0.7 g/kg dw), lipids (301.2+11.9 g/kg dw), carbohydrates (243.9+9.9g/kg dw) and fibers (313.2 g/kg dw). The HPLC profile revealed the presence of polyphenols (21.2 + 1.18 mg eq. QE/g Ext.), flavonoids (56.4 + 6. 47 mg eq. QE/g Ext.) and phenolic acid like caffeic and coumaric acidss. Iron was the most micronutrients found but we also found copper, manganese, zinc selenium. The percentage of phycocyanin was 16.15% while carotenoids were 3.8%.
Conclusion: S. platensis from Nomayos have significant concentration of nutrients. However other studies need to be carried out in order to determine the nature of different monomers present in protein, lipids, carbohydrates and other macronutrients found in this microalgae.
Keys words : spirulina, chemical composition, phycocyanin, polyphenols.

Full text

Annals. Food Science and Technology
2016
Available on-line at www.afst.valahia.ro
524
Volume 17, Issue 2, 2016
CHEMICAL COMPOSITION OF SPIRULINA PLATENSIS OF NOMAYOS-YAOUNDE
(CAMEROON)
Vicky Jocelyne Ama Moor1*, Constant Anatole Pieme1, Prosper Cabral Nya Biapa2 Marthe Elise Ngo
Matip3, Bruno Moukette Moukette 1, Francine Tankeu Nzufo 1, Pauline Nanfack4, Jeanne Ngogang1.
1Faculty of Medicine and Biomedical Sciences- University of Yaounde 1 – Cameroon. PO. Box 1364
2 Faculty of Sciences, University of Dschang – Cameroon
3 Faculty of Sciences, University of Ngaoundere - Cameroon
4 IMPM, University of Yaounde 1 – Cameroon
*Corresponding author: movicky@yahoo.fr
ABSTRACT
Background: S. platenis is rich in macronutrients and micronutrients. It is used as a dietary supplement and found in
tropical and semi-tropical areas This study aims to investigate the chemical composition of S. platensis of Nomayos
(Yaounde, Cameroon) focusing on macro and micronutrients and phyotchemical bioactive molecules
Materiel and methods: S. platensis was collected in a farm in Nomayos (Yaounde, Cameroon). The aqueous extract
was used for the determination of nutrients using standard methods. Trace elements were determined by atomic
absorption spectrophotometry. The phytochemical composition was determined by using HPLC. Phycocyanin and
Carotenoids were determined using chemicals methods.
Results: The extraction yield obatined was 16.84%. The results showed that S platensis contains protein (375.5+0.7
g/kg dw), lipids (301.2+11.9 g/kg dw), carbohydrates (243.9+9.9g/kg dw) and fibers (313.2 g/kg dw). The HPLC
profile revealed the presence of polyphenols (21.2 + 1.18 mg eq. QE/g Ext.), flavonoids (56.4 + 6. 47 mg eq. QE/g Ext.)
and phenolic acid like caffeic and coumaric acidss. Iron was the most micronutrients found but we also found copper,
manganese, zinc selenium. The percentage of phycocyanin was 16.15% while carotenoids were 3.8%.
Conclusion: S. platensis from Nomayos have significant concentration of nutrients. However other studies need to be
carried out in order to determine the nature of different monomers present in protein, lipids, carbohydrates and other
macronutrients found in this microalgae.
Keys words : spirulina, chemical composition, phycocyanin, polyphenols.
Submitted: 26.09.2016 Reviewed: 11.11.2016 Accepted: 16.12.2016
1. Introduction
Spirulina platensis (Oscillatoriaceae) is a blue-
green microalgae that has been found by
several scientists and nutritionists to be a
functional food with exceptional nutritional
qualities (FAO, 1991). The cultivation of S.
platensis is widespread all over the world with
high productivity and low cost of production
(Charpy , Langlade, & Alliod , 2008). This
algae preferably grows in warm alkalinated
waters rich in nitrogen and phosphorus
nutrients such as in some of the lakes in Africa,
latin America, and South Asia. In addition it is
found in tropical and semi-tropical areas
(Castenholz & al., 2001). The thermophilic
character of S. platensis and its important needs
of light limit its range to a tropical belt between
approximately 35° north and 35 ° south
latitude. S. platensis’ production takes place at
several levels: artisanal, semi-industrial and
industrial. The differentiating factors of these
production methods are the total area of
cultivation tanks, the equipment, and
technology used in the production process
(IRD, 2008). As mentioned above, S. platensis
is present in various part of the world, and
depending on its location, it takes on a different
name. For instance in the far North region of
Cameroon it is called sembe’’; in Chad it is
called ‘’dihe’’. Rich in proteins, minerals, trace
elements, and many vitamins such as B1, B2,
B12 and E, this microalgae is considered an
unconventional food resource (Sall & al.,1999).
In Cameroon it is produced and used as a
functional food, however no data has been
published regarding its composition. Therefore
our study aims to investigate the micro and
macronutrients composition of S. platensis
produced at Nomayos in Cameroon.
 2016 Valahia University Press
Further reproduction without permission is prohibited

Annals. Food Science and Technology
2016
Available on-line at www.afst.valahia.ro
525
Volume 17, Issue 2, 2016
2. Material and methods
Spirulina platensis was provided from a farm
situated at Nomayos (Yaounde- Cameroon).
The algae was cultivated inside a basic water
tank with pH 10. Each day a third of the culture
medium was collected on a fabric filter which
passed water while retaining the spirulina
accumulated on the very fine mesh filter (30 to
40μ). This spirulina was then pressed for 20
minutes to obtain a biomass which was spread
out on racks in the form of spaghetti. Next it
would be dried in an electric dryer at 40 °C, a
temperature that will ensure a better
preservation of vitamins and phytonutrients.
The dried algae was then grinded into a fine
powder.
2.1. Preparation of the aqueous extract
The aqueous extract was obtained by
macerating 100 g of S. platensis powder with
1000 mL of distilled water. This mixture was
then placed on intermittent agitation for 24
hours, and the filtrate obtained was
immediately lyophilized.
2.2. Determination of macronutrients and
micronutrients
The determination of total ash protein,
carbohydrates, lipids, and fibers were
determined using standard methods (Van Soest,
1967; AOAC, 1980; AOAC, 1984; Bergeret,
1985). The determination of micronutrients
was carried out using atomic absorption
spectrophotometry after extraction in a mixture
of nitric- hydrochloric acid (75v/ 25v).
2.3. Phytochimical screening and phenolic
profile of S.platensis
2.3.1. Determination of the phenolic content
of the extract
The total phenol, flavonoids, and flavonols
contents were determined by standard methods
(Singleton & Rossi, 1965; Zhishen & al., 1999;
Kumaran & Karunakaran, 2006).
2.3.2.Total antioxidant activity by Ferric
Reducing Antioxidant Power assay (FRAP)
The Total antioxidant activity was determined
using documented method by Benzie and
Strain (Benzie & Strain, 1996) with some
modifications (Moukette & al., 2015). The
fresh solution of FRAP reagent consisted of
acetate buffer (300 mM; pH 3.6), 2,4,6- Tri (2-
pyridyl)-s-triazin (TPTZ) (10 mM), and
FeCl3•6HO (50 mM) in the ration 10v:1v:1v.
For the assay 75 µL of S. platensis extract and
2 ml of FRAP reagent were mixed for 12
minutes. The absorbance of this mixture was
measured at 593 nm. . The vitamin C was used
to draw a standard curve and the results were
expressed as mg equivalent vitamin C/ g of
dried extract (mg eq VitC/g DE).
2.3.3.Quantification of phenolic compounds
by HPLC
High Performance Liquid Chromatography
(HPLC) with UV detection was used for
identification and detection of phenolic profile
of the extract. The solution of S. platensis
extract was dissolved in pure water in the ratio
of 30 mg extract/1mL water and centrifuged at
4706 rpm for 10 minutes. The supernatant was
filtered through a cellulose acetate membrane
filter 0.20 µm - 0.45µm, (Schleicher &
Schuell). The analysis of the filtrated solution
was performed on an Agilent Technologies
1200 HPLC system fitted with a SUPELCOSIL
LC-18 column (length 250 mm, diameter 4.6
mm, packaging size 5 mm). The column
temperature was set at 20°C. The mobile phase
was made of a mixture of solution of acetic
acid 0.5% by volume (A) and acetic nitrile (B).
The elution was performed by using 100% of A
for the first 2 minutes of the run, 40% of A and
60% of B from 2 to 60 minutes. The flow rate
was set equal to 1 mL/min and the volume 25
µL was injected in the column. Polyphenols
were detected by a UV detector (280 nm). The
retention times of the identified different
phenolic compounds were measured using a
single standard solution (100 mg/L). This
method was also previously used by Moukette
and al (Moukette & al., 2015).
2.4.Determination of phycocyanin and
carotenoids
For the dermination of phycocyanin, 3g of S.
platensis was diluted with 100mL of distilled
water and decanted. Then 20mL of supernatant
were centrifuged (3000 rpm, 10 minutes) and
0.5 mL of the supernatant was diluted 100
times with distilled water. The optical density
(OD) of each sample was

Annals. Food Science and Technology
2016
Available on-line at www.afst.valahia.ro
526
Volume 17, Issue 2, 2016
spectrophotometrically measured at 615 and
652 nm (Jourdan, 2006).
Regarding the investigation of carotenoids
concentration in the S. platensis extract, 3 g of
S. platensis powder were extracted with 25 mL
of acetone; the mixture was kept for 24h in the
fridge. The surpernatant was centrifuged at
3000 rpm for 10 minutes and 0.5 mL of it was
diluted 100 times. The OD was read at 450 nm
(Vonshak & Borowitzka, 1991).
3. Results And Discussion
The extraction yield obatined was 16.84% and
was calculated using the following formula: Rd
= Mass of the extract obtained × 100 / initial
powder mass. The percentage of humidity was
07.24%+0.26. The nutritional composition of
S. platensis presented in table 1 showed the
presence of high level of protein and iron. S.
platensis of Nomayos demonstrated a lower
concentration of protein and some trace
elements compared to the one harvested in
Switzerland (65% of protein), Burkina Faso
(61.3 % of protein) and Chad (58.61% and
50.24% of protein) (Azabji & al., 2011;
Branger & al., 2003; Ngakou & al., 2012). The
differences could be explained by either the
growth environment, the difference in climate,
or the techniques used to collect S. platensis
(Clement, 1975). The protein content in this
algae could vary with the sampling period in
relation to the photoperiod with the highest
contents being obtained at the beginning of the
sunlight period (AFAA, 1982). S. platensis of
Nomayos showed more lipids than the one
harverted in Chad (Ngakou & al., 2012). This
difference could be due to a variation of the
extraction method or the type of solvent used.
Furthermore our study revealed that the level of
iron was 25.6 mg/100g DM. This is higher
compared to that found in other studies (0.7 to
1.2 mg/100g DM) (Montasell, 2009).
The phytochemical screening revealed the
presence of polyphenols (56.4 + 6.47 mg eq.
QE/g Ext.), total flavonoids (21.2 + 1.18 mg
eq. QE/g Ext.), total flavonols (13.2 + 0.6 mg
eq. QE/g Ext.); and total antioxidant capacity
(7.5 + 0.33 mg eq. VitC/g Ext.). The phenolic
profile of S. platensis using high performance
liquid chromatography showed the presence of
several phenolic acids especially caffeic,
coumaric O and P, gallic, and other bioactive
molecules at different levels (Table 2). Our
results are similar to other studies which
demonstrated the presence of phenolic and
other bioactive molecules like phycocyanin
(Bhavisha & Parula, 2010). We obtained
16.15% of phycocyanin which is more than the
15% obtained by Fox (1986) and Jourdan
(1999). The carotenoid pigments including ß-
carotene are important as the main source of
vitamin A represented 3.80%.
Table 1: Nutritional composition of S. platensis of Nomayos -Yaounde
Macro-
nutrients
(%)
Dry weight
Lipids
Proteins
Fibers
Sugars
Energy
92,76+0.26
30.12+1.19
37.55+0.07
31.32+7.95
24.39+0.99
518.84 kcal
Micro-
nutrients
(mg /Kg)
Iron
Manganese
Copper
Zinc
Selenium
Ash
256.56+ 0.01
23.38+0.00
28.95+0.00
25.01+0.01
1.24+0.01
07.93+0.20
Table 2: HPLC polyphenols profile of S. platensis
Polyphenol standard
Standard Retention time
Spirulina platensis
TR(min)
Con [mg/L]
A(mAU) (mg/g DW)
Gallic
14.38
422.87
1.65x104
3,4-OH benzoic
19.10
41.92
1062.86
Tyrosol
21.76
40.91
700.29
OH-Tyrosol
21.906
70.15
759.48
Catechine
23.48
66.84
919.68
O-Coumaric
25.10
140.87
4409.89
Caffeic
25.66
518.63
1.68x104
P-Coumaric
30.51
201.68
1.03e4
Apigenin
33.49
1.66
7333.82
Conc: concentration; DW: dried weight; T.R.: retention time; A: area

Annals. Food Science and Technology
2016
Available on-line at www.afst.valahia.ro
527
Volume 17, Issue 2, 2016
The diversity of micro and macronutrients
found in S. platensis increased the focus on this
algae and its used as a marketed food
supplement in many countries in Europe,
Africa, Asia and India. Studies demonstrated
important properties of S. platensis such as an
antioxidant, antiviral, antibacterial, antidiabetic
and other biological activities (Lee & al., 2001;
Bhavisha & Parula, 2010). Recent studies also
demonstrated benefit effects of S. plantensis
from Nomayos on patients living with
HIV/AID by increasing the level of CD4 cells
count, reducing the viral loads and increasing
HDL levels (Ngo Matip & al., 2014; Ngo
Matip & al., 2015). The benefits from S.
platensis could be attributed to either its
micronutrients or to other bioactive molecules
in the algae.
4. Conclusion
Given its chemical composition and richness in
micro and macronutrients, as well as other
bioactive molecules, we can attest to the
benefits of S. platensis from Nomayos.
However other studies need to be carried out in
order to determine the nature of different
monomers present in protein, lipids,
carbohydrates and other macronutrients found
in this microalgae.
Author’s contribution: VJAM and CAP
conducted the study; PCNB, BMM, TNF and
PN did the analysis and carried the study,
MENM gave S. platensis and JN supervised the
study. All the authors approve the content of
the work.
Conflicts of interest: The authors declare no
conflict interest
Funding: This research did not receive any
specific grant from funding agencies in the
public, commercial, or not-for-profit sectors.
References
[1] AFAA (French Association for Applied Algology).
First symposium on Spirulina platensis Gom. Geitleir of
AFAA 1982.
[2] AOAC. Official methods of analysis of the
association of official analytical chemists(13rd ed)
William Horturitz ed. Washingtoon DC. 1980.
[3] AOAC. Official methods of analysis association of
official analytical chemists. Centennial edition 1884 –
1984 (14 th ed), Virginia 22209 USA. 1984
[4] Azabji, K. M., Loni, G.E., Sobngwi, E., Onana,
E. A., Edie, D.S., Denis von der Weid, & al. The Effect
of Spirulina platensis versus Soybean on Insulin
Resistance in HIV-Infected Patients: A Randomized
Pilot Study. Nutrients 2011; 3 :712-724
[5] Benzie, I.F.F. & Strain, J.J (1996). Ferric
reducing ability of plasma (FRAP) as a measure of
antioxidant power: The FRAP assay. Analytical
Biochemistry 1996 ; 239:70-76
[6] Bergeret, B. La chimie analytique dans un
laboratoire d’outre-mer. Organisation des techniques de
laboratoire : étude critique des techniques d’analyses
employées au laboratoire de la section nutrition de
l’IRCAM – ORSTOM, IRCAM. 56p 1955.
[7] Bhavisha, R. & Parula, P. Spirulina : Potential
clinical therapeutic application. Journal of Pharmacy
Research 2101 ; 3(8) :1726-1732
[8] Branger, B., Cadudal, J.L., Delobel, M., Ouoba,
H., Yameogo, P., Ouedraogo, D., Guerin, D., Valea
,A., Zombre, C. & Ancel, P.; personnels des CREN
(2003). Spiruline as a food supplement in case of infant
malnutrition in Burkina-Faso. Archives de Pediatrie
2003 ; 10(5):424-31
[9] Castenholz, R.W., Rippka, R., Herdman, M. &
Wilmotte, A. Form-genus I. Arthrospira 2001.
[10] Clement, G. Production et constituants
caractéristiques des algues Spirulina platensis et
maxima. Annales de la Nutrition et de l’ Alimentation
1975 ; 29 : 447-488.
[11] Charpy, L., Langlade, M.J. & Alliod, R. « La
Spiruline peut-elle être un atout pour la santé et le
développement en Afrique ? » ; IRD : 67 P. 2008.
[12] Cornet, J.F.Cinétic and energetic study of aphoto-
bioreactor. Doctorate Thesis, University de Paris-South
Orsay Centre. In: for de spirulina production. Jourdan,
J.P (Ed.). Antenna Technology, 146pp. 1992.
[13] FAO/WHO/UNU. Protein Quality Evaluation.
Report of the Joint FAO/WHO ExpertConsultation:
1985 FAO/WHO/UNU 2- to 5-yearold requirement
pattern. FAO; 51. 1991.
[14] Fox , R.D. Algoculture : la spiruline, un espoir
pour le monde de la faim. Edisud, Aix-en-Provence,
ISBN 2-85744-262-9, 319 p.1986.
[15] Jourdan, J.P. Grow your spirulina. In: Jourdan,
J.P. (Ed). Antenna Technology. 146pp. 2006.
[16] Kumaran, A. & Karunakaran, R.J. Nitric oxide
radical scavenging active components from Phyllanthus
emblica L. Plant Foods for Human Nutrition 2006 ;
61(1):1-5.
[17] Lee, J.B., Srisomporn, P., Hayashi, K.,Tanaka,
T., Sankawa, U. & Hayashi, T. Effects of Structural
Modification of Calcium Spirulan, a Sulfated
Polysaccharide from Spirulina Platensis, on Antiviral
Activity. Chemical and Pharmaceutical Bulletin 2001 ;
49(1) : 108—110

Annals. Food Science and Technology
2016
Available on-line at www.afst.valahia.ro
528
Volume 17, Issue 2, 2016
[18] Mitchell, G.V., Grundel, E., Jenkins, M. &
Blakely, S.R. Effects of graded dietary levels of
Spirulina maxima on vitamins A and E in male rats.
Journal of Nutrition, 1990 ; 120(10): 1235-40
[19] Montasell. Montasell-Import SARL,
Spirulina,Pressbook, p. 12-13. 2009.
[20] Moukette Moukette, B., Pieme, C.A., Nya Biapa,
P.C., Njimou, J.R. & Ngogang Yonkeu, J. Free
radicals quenching potential, protective properties
against oxidative mediated ion toxicity and HPLC
phenolic profile of a Cameroonian spice: Piper
guineensis. Toxicology Reports 2015 ; 2:792–805.
[21] Ngakou, A., Ridine, W., Mbaïguinam, M. &
Namba, F. Changes in the physico-chemical properties
of Spirulina platensis from three production sites in
Chad. Journal of Animal & Plant Sciences 2012 ; 13(3):
1811-1822
[22] Ngo-Matip, M.E., Pieme, A.C., Azabji-Kenfack,
M., Nya Biapa, P.C., Nkenfack, G., Heike, E.,
Moukette Moukette, B., Korosky, E.l., Mbofung,
C.M. & Yonkeu Ngogang, J. Effects of Spirulina
platensis supplementation on lipid profile in HIV–
infected antiretroviral naïve patients in Yaounde -
Cameroon: a randomized trial study. Lipids in Health
and Disease 2014 ; 13:191
[23] Ngo-Matip, M.E., Pieme, C.A., Azabji-Kenfack,
M., Moukette Moukette, M., Korosky, E., Stefanini,
P., Yonkeu Ngogang, J. & Mbofung, C.M. Impact of
daily supplementation of Spirulina platensis on the
immune system of naïve HIV-1 patients in Cameroon: a
12-months single blind, randomized, multicenter trial.
Nutrition Journal 2015 ; 14:70
[24] Pinero Estrada, J.E., Bermejo Besco´, P. &
Villar del Fresno, A.M. Antioxidant activity of different
fractions of Spirulina platensis protean extract. Il
Farmaco 2001 ; 56 :497–500
[25] Sall, M.G., Dankoko , B., Badiane, M., Ehua, E.
& Kuakuwi, N. Résultats d'un essai de réhabilitation
nutritionnelle avec la Spiruline à Dakar. Médecine
d'Afrique Noire 1999 ; 46:143-14
[26] Singleton, V. & Rossi, J. Colorimetry of total
phenolics with phosphomolybdic phosphotungstic acid
reagents. American Journal of Enology and Viticulture
1965 ; 16:144–58.
[27] Stizenberger. Bergey's Manual of Systematic
Bacteriology (D. R. Boone & R.W.
[28] Castenholz, eds.) 1852 ; 1: 542-54
[29] Van Soest, R.J. & Wine, R.H. JAOAC (50)
Washington DC. 1967.
[30] Vonshak, A. & Borowitzka, M.A. Laboratory
Manual: Research Seminar and Workshop on Mass
Cultures of Microalgae. Silpakorn University, Thailand.
40pp. 1991.
[31] Zhishen, J., Mengcheng, T. & Jianming, W. The
determination of flavonoid contents in mulberry and
their scavenging effects on superoxide. Radicals Food
Chemistry 1999 ; 64:555–9.