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In . J. Biosci.
2020
RESEARCH PAPER OPEN ACCESS
Nu i ional alues o
Spi ulina pla ensis
biomass cul i a ed in
Eas A ica
Fe en Teze a1*, Musa Chacha2, Ma y John3, Jo ey Raymond1
1Depa men o Food and Nu i ional Sciences, School o Li e Science and Bioenginee ing, The
Nelson Mandela A ica Ins i u ion o Science and Technology, A usha, Tanzania
2Depa men o Sus ainable Ag icul u e and Biodi e si y Conse a ion, School o Li e Science and
Bioenginee ing, The Nelson Mandela A ica Ins i u ion o Science and Technology, A usha,
Tanzania
3Depa men o Heal h and Biomedical science, School o Li e Science and Bioenginee ing, The
Nelson Mandela A ica Ins i u ion o Science and Technology, A usha, Tanzania
Key wo ds: Spi ulina pla ensis, Nu i ional alue, Phy ochemical, Eas A ica.
h p://dx.doi.o g/10.12692/ijb/16.6.121-128
A icle published on June 16, 2020
Abs ac
Spi ulina pla ensis is a biomass o cyanobac e ia adi ionally used o hund ed yea s wo ldwide o i s nu i ional and
pha macological bene i s. So a , his mic oscopic o ganism has no unde gone many scien i ic s udies in Eas A ica.
The e o e, e alua ing i s nu i ional p ope ies could help o jus i y i s po en ial and p omo e i s u iliza ion in he egion. In
his s udy, he de e mina ion o i amins was pe o med by using a high-pe o mance liquid ch oma og aphy (HPLC) me hod.
Analysis o mine als and hea y me als was pe o med by using a omic abso p ion spec opho ome e (AAS). The p o ein
con en o Spi ulina pla ensis was de e mined by using he Kjeldahl me hod. The spec opho ome y me hod was used o
phy ochemical analysis. The esul s showed ha he i amin A, i amin B9 and i amin B12 con en o Spi ulina pla ensis was
27.4 𝜇g/100g, 246.89 𝜇g/100g, and 3.87 𝜇g/100g espec i ely. Concen a ions o i on, zinc, calcium, and phospho ous in
spi ulina pla ensis we e ound o be 82.2 mg/100g, 85.1 mg/100g, 1300 mg/100g, and 600 mg/100g espec i ely. The p o ein
con en o spi ulina pla ensis was 46 g/100g. The concen a ions o o al phenolic and o al la onoid in spi ulina pla ensis
ound o be 2.99 mg GAE/g and 1.92 mg QE/g espec i ely. These da a indica ed ha he nu ien concen a ion o spi ulina
pla ensis is enough o mee he ecommended daily allowance (RDA) o mos essen ial nu ien s. Me cu y, lead, cadmium and
a senic con en s o spi ulina pla ensis was 0.000036 mg/kg, 0.0047 mg/kg, 0.00048 mg/kg and 0.0047 mg/kg espec i ely.
All he analyzed hea y me als we e wi hin he ecommended EU sa e y limi s. Ou indings con i med he nu i ional po en ial
o Spi ulina pla ensis cul i a ed in Eas A ica. Thus, Spi ulina pla ensis could be conside ed as a po en ial al e na i e sou ce
o sus ainable nu i ion o add ess malnu i ion in he egion.
* Co esponding Au ho : Fe en Teze a damessa @nm-ais .ac. z
In e na ional Jou nal o Biosciences | IJB |
ISSN: 2220-6655 (P in ), 2222-5234 (Online)
h p://www.innspub.ne
Vol. 16, No. 6, p. 121-128, 2020
122
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In . J. Biosci.
2020
In oduc ion
Spi ulina is a genus o blue-g een algae, which
belongs o phylum cyanobac e ia. Spi ulina has
di e en species howe e , Spi ulina pla ensis and
Spi ulina maxima a e he wo commonly consumed
species by humans and animals. Nume ous s udies
epo ed ha spi ulina is he iches sou ce o high-
quali y p o ein, i amins, mine als, and o he heal h-
p omo ing bioac i e compounds ha a e mos ly
inadequa e in ypical A ican die s (Kay and Ba on,
2009; B una e al., 2015; Saha and Mu ay, 2018).
Besides, unlike o he ood c ops, spi ulina equi es
less space and inpu s o g ow (Ga cía e al., 2017;
Capo gno and Ma hys, 2018; Khan e al., 2018).
Spi ulina h i es in alkaline lakes whe e i is di icul
o impossible o o he mic oo ganisms o su i e.
Lakes in Eas A ica along he G ea Ri Valley
including Lake Na on, Lake Tanganyika, Lake
Naku u, Lake Elemen ei a and also Lake Vic o ia
ha e he special cha ac e is ic ha makes hem
a o able o spi ulina p oduc ion (Habib e al., 2011;
An onio, 2011).
Al hough Eas A ica has he mos a o able
en i onmen al condi ion, p oduc ion, as well as
u iliza ion o spi ulina, is e y limi ed in he egion;
besides, he e is limi ed o no scien i ic in o ma ion
on he nu ien and phy ochemical composi ion and
o he nu i ional p ope ies o spi ulina g own in
Eas A ica. Hence, we belie ed ha he exis ence o
limi ed scien i ic in o ma ion es ic ed he wide
p oduc ion and u iliza ion o spi ulina in Eas A ica.
The e o e, his s udy aimed o p o ile he nu ien
composi ion and o he nu i ional p ope ies o
locally cul i a ed Spi ulina pla ensis biomass.
Ma e ials and me hods
Spi ulina samples
The powde o Spi ulina pla ensis used in his s udy
was ob ained om local p oduce s om di e en
a eas o Kenya. Spi ulina pla ensis was cul i a ed in a
pond co e ed wi h a g eenhouse. A e ha es ing, he
biomass o spi ulina pla ensis was d ied and
g ounded o ob ain powde p oduc s.
De e mina ion o i amins
The de e mina ion o i amins was pe o med using
HPLC ollowing he me hods desc ibed by Sami e al.
(2014). Fo i amin A, 1 g o py ogallic acid, 70 mL
e hanol, and 30 mL (50%) KOH we e added in o 10 g
o powde spi ulina pla ensis, s i ed and e luxed o
40 minu es using a wa e ba h a 50 °C. Ex ac s we e
ob ained h ee imes using 50 mL, 30 mL, and 20 mL
e he concen a ions. Dis illed wa e was used o
neu alize he ex ac . Fu he , he ex ac was
concen a ed o app oxima ely 5 mL by using a wa e
ba h (50 °C), dilu ed o 10 mL by using me hanol,
il e ed using a 0.45 𝜇m memb ane, and inally
subjec ed o HPLC analysis.
To de e mine i amin B9 and B12, abou 2 g powde
spi ulina pla ensis was placed in 25 mL o sul u ic
acid (0.1 N) solu ion and incuba ed o 30 minu es a
121°C. Then, he con en was cooled and adjus ed o
pH 4.5 wi h 2.5 M sodium ace a e, and 50 mg
Takadias ase enzyme was added. The p epa a ion was
s o ed a 35°C o e nigh . The mix u e was hen
il e ed h ough a Wha man No. 4 il e , and he
il a e was dilu ed wi h 50 mL o pu e wa e and
il e ed again h ough a mic opo e il e (0.45 𝜇m).
Twen y mic oli e s o he il a e we e injec ed in o
he HPLC sys em. Quan i ica ion o i amin B9 and
B12 con en was accomplished by compa ison o
s anda ds.
De e mina ion o mine als and hea y me als
Concen a ions o mine als and hea y me als we e
de e mined using a omic abso p ion
spec opho ome e (AAS) modi ying a me hod
desc ibed by S eponënienë and Tau kus (2006).
De e mina ion o mine als and hea y me als was
ca ied ou a e he p epa a ion o samples and
s anda d solu ions. Calib a ion solu ions we e used
o he calib a ion o he a omic abso p ion
spec opho ome e . The sample was p epa ed by
adding 10 g powde spi ulina pla ensis in o a 250 mL
olume ic lask and mixed wi h 10 mL o ni ic acid.
The mix u e was hea ed o 10 minu es using Kjeldahl
block diges e . A e cooling i down, 5 mL o ni ic
acid was added and hea ed again o 30 minu es.
123
Teze a e al.
In . J. Biosci.
2020
A e lea ing he solu ion o cool o 10 minu es, 2 mL
dis illed wa e , 3 mL hyd ogen pe oxide, and 2 mL
hyd ochlo ic acid was added and hea ed again o
ano he 10 minu es. The solu ion was il e ed using
Wha man il e pape 1 and dilu ed o 100 mL using
dis illed wa e . Readings by a omic abso p ion
spec opho ome e we e done a di e en
wa eleng hs; 248.3 nm, 213.9 nm, 422.7 nm, 213.6
nm, 193.7 nm, 228.8 nm, 253.7 nm, 283.3 nm was
used o i on, zinc, calcium, phospho us, a senic,
cadmium, me cu y and lead espec i ely.
De e mina ion o phy a e
Phy a e was de e mined using an anion-exchange
me hod ollowing Ma e al. (2005). Samples we e
accu a ely weighed (1.0-2.0 g) and ans e ed in o
100 mL conical lasks. A o al o 40-50 mL o Na2SO4
(100g/L)-HCl (1.2%) was added. Flasks we e hen
capped and shaken igo ously o 2 hou s on a o a o
a ambien labo a o y empe a u e.
The supe na an was hen il e ed h ough quali a i e
il e pape no 4. A o al o 10 mL o il e ed ex ac
was dilu ed o 30 mL wi h dis illed wa e a e mixing
wi h 1 mL o 0.75M NaOH and hen passed h ough
an anion esin column ( esin AG1-X4, ~ 100-
200mesh, Bio ad Labo a o y Inc., column 0.8 x
10cm). The column was washed be o e use wi h 20
mL o 0.5 mol/L NaCl solu ion and deionized wa e .
A e sample applica ion, he column was washed
wi h 15 mL o dis illed wa e and 20 mL o 0.05M
NaCl solu ion o emo e he ino ganic phospha e.
Then he e ained phy ic acid was elua ed wi h 0.7M
NaCl.
The pos -column eagen was made up o a0.03%
FeCl3 solu ion con aining 0.3% sul osalicylic acid. A
o al o 4 mL o he eagen was added in o 5 mL o
collec ed elua e and cen i uged a 3000 pm o 10
minu es. The abso bance o he supe na an was
measu ed a 500 nm using a spec opho ome e (LKB
4053, U.K.). A calib a ion cu e o he colo ime ic
me hod was ob ained by using phy a e s anda ds (P-
8810 Sigma Co.). The phy a e con en o samples was
calcula ed using he s anda d cu e.
De e mina ion o p o ein
The Kjeldahl me hod desc ibed by Mæh e e al.
(2018) was used o de e mine spi ulina p o ein
con en . B ie ly, a powde sample o 1 g was measu ed
and added o a diges ion lask along wi h 25 mL
sul u ic acid, 15 g po assium sul a e, and 0.3 g coppe
(II) oxide. The mix u e was hea ed o 2 hou s a 370
°C. A e cooling i down, 300 mL dis illed wa e was
added. The diges ed sample was connec ed o he
dis illa ion appa a us and he dis illa e was collec ed
in o he collec ing lask which con ains 25 mL sul u ic
acid (HCL), 150 mL dis illed wa e , and a ew d ops o
mixed indica o . The dis illed solu ion was i a ed
wi h sodium hyd oxide solu ion (0.1 mol/L) un il he
colo o he solu ion is changed om iole o g een.
The olume was eco ded a he poin whe e he colo
is changed.
De e mina ion o phy ochemicals
The o al phenolic (TPC) and la onoid (TFC)
con en s we e de e mined using a spec opho ome e
ollowing he me hod as desc ibed by Chand a e al.
(2014) wi h sligh modi ica ions. In b ie , powde
spi ulina (10 g) om each sample was ex ac ed using
75 mL (95% / ) me hanol a 40°C o 10 minu es
ollowed by cen i uga ion a 3500 pm o 10
minu es. The clea supe na an was collec ed and
s o ed in an ambe bo le o analysis.
P epa a ion o gallic acid s anda ds
Va ious concen a ions o Gallic acid solu ions in
me hanol (5-500 mg/L) we e p epa ed. In a 20 ml
es ube, 1 mL Gallic acid o each concen a ion was
added and o ha 5 mL o Folin-Ciocal eu’s eagen
(10%) and 4 mL o 7% sodium ca bona e we e added
o ge a o al olume o 10 mL. The blue-colo ed
mix u e was shaken well and incuba ed o 30
minu es a 40 0C in a wa e ba h. Then he
abso bance was measu ed a 765 nm agains a blank
(0.5 mL me hanol, 2.5 mL 10% FolinCiocal eu’s
eagen , and 2.5 mL o 7% sodium ca bona e). All he
expe imen s we e ca ied ou in iplica e. The
a e age abso bance alues ob ained a di e en
concen a ions o Gallic acid we e used o plo he
calib a ion cu e.
124
Teze a e al.
In . J. Biosci.
2020
To al phenolic con en
The o al phenolic con en in spi ulina sample was
de e mined by using he Folin and Ciocal eu eagen
me hod. The es samples we e p epa ed by mixing
0.5 mL o ex ac , 2.5 mL o 10% Folin-Ciocal eu’s
eagen , and 2.5 mL 7% sodium ca bona e. The
samples we e he ea e incuba ed a 45 °C o 45
minu es in a wa e ba h. The phenolic con en was
calcula ed as Gallic acid equi alen s GAE/g o d y
sample based on a s anda d cu e o Gallic acid (5-
500 mg/L, 𝑌 = 0.5129𝑥 + 0.0145, 𝑅2 =0.9985).
To al la onoids con en
The o al la onoid con en was es ima ed acco ding
o he aluminum chlo ide colo ime ic me hod. Fo
o al la onoid de e mina ion, que ce in was used o
make he s anda d calib a ion cu e. The s ock
que ce in solu ion was p epa ed by dissol ing 50 mg
que ce in in 10 mL me hanol. Que ce in s anda d
cu e was buil using a concen a ion o 50–250
𝜇g/mL. An amoun o 0.6 mL dilu ed s anda d
que ce in solu ions and he same amoun o ex ac
was sepa a ely mixed wi h 0.6 mL o 2% aluminum
chlo ide. A e mixing, he solu ion was incuba ed o
60 minu es a oom empe a u e. The abso bance o
he eac ion mix u es was measu ed a 420 nm. Wi h
a Va ian UV-Vis spec opho ome e .
The concen a ion o o al la onoid con en in he
es samples was exp essed as mg que ce in
equi alen (QE)/g o d y sample based on a s anda d
cu e o que ce in (50-250 𝜇g/mL, 𝑌 = 0.0017𝑥 -
0.0089, 𝑅2 =0.9982).
S a is ical analysis
Da a we e analyzed using a s a is ical so wa e IBM
SPSS e sion 23. All he desc ip i e da a we e
p esen ed as mean alues.
Resul s
Resul s showed ha 100 g o spi ulina pla ensis
g own in Eas A ica con ains a signi ican amoun o
essen ial nu ien s and bioac i e compounds ha a e
impo an o human nu i ion. As desc ibed in
Tables 1 and 2, 100 g o spi ulina pla ensis g own in
Eas A ica con ains mine als, i amins, and p o ein
in a concen a ion enough o mee he ecommended
daily allowance (RDA) o a young child.
Table 1. Vi amin A, i amin B9, i amin B12, and p o ein composi ion o Spi ulina pla ensis.
Nu ien
Concen a ion
Vi amin A (𝜇g/100g)
27.4
Vi amin B9 (𝜇g/100g)
246.89
Vi amin B12 (𝜇g/100g)
3.87
P o ein (g/100g)
46
The s udy u he e ealed ha spi ulina pla ensis
has a e y low concen a ion o hea y me als (Table
5). All he analyzed hea y me als we e wi hin he
ecommended EU sa e y limi s. Also, as desc ibed in
Table 3, he a io o mine als o phy a e a e wi hin he
anges ha a o mic onu ien s bioa ailabili y. As
summa ized in Table 4, spi ulina pla ensis also
con ains a signi ican amoun o well-known heal h-
p omo ing an ioxidan s such as
phenolic compounds and la onoids.
Discussion
In his s udy, we p o iled he nu ien and
phy ochemical composi ion o locally p oduced
spi ulina pla ensis o e alua e i s po en ial o u ilize i
as an al e na i e ood supplemen o add ess
unde nu i ion in Eas A ica. Ou indings showed
ha spi ulina pla ensis cul i a ed in Eas A ica a e
ich in essen ial nu ien s. Mo eo e , a small amoun
o spi ulina pla ensis is su icien o mee he
ecommended die a y allowance (RDA) o mos
nu ien s (Ins i u e o Medicine, 2001). The analyzed
100 g spi ulina pla ensis powde is su icien o mee
he ecommended die a y allowance (RDA) o
p o ein, i on, zinc, calcium, and i amin B12 o
indi iduals in all age g oups.
125
Teze a e al.
In . J. Biosci.
2020
Table 2. Concen a ions o i on, zinc, calcium, phospho us, and phy a e in Spi ulina pla ensis (mg/100g).
Nu ien
Concen a ion (mg/100g)
I on
82.8
Zinc
85.1
Calcium
1300
Phospho ous
600
Phy a e
1.86
The same amoun o spi ulina pla ensis could p o ide
su icien phospho us o mee he ecommended
die a y allowance (RDA) o 0-8 yea s old child en.
Mo eo e , a 100 g spi ulina pla ensis is enough o
p o ide 48% and 86% phospho ous o mee he
ecommended die a y allowance (RDA) o 9-18 and
19-50 yea s old indi iduals espec i ely. The
ecommended die a y allowance (RDA) o i amin B9
o child en aged 0-8 yea s can be su icien ly me by
only 100 g spi ulina pla ensis; abou 100 g o
spi ulina pla ensis is enough o p o ide mo e han
50% o he ecommended die a y allowance (RDA) o
i amin B9 o indi iduals aged 9 o 50 yea s. On he
o he hand, a 100 g spi ulina pla ensis can p o ide
only 9% o he ecommended die a y allowance
(RDA) o i amin A.
Table 3. Phy ic acid o i on, phy ic acid o zinc, and phy ic acid o calcium mola a ios.
Sample
Phy a e: Fe
Phy a e: Zn
Phy a e: Ca
Spi ulina powde
0.0019
0.0021
0.000086
Sugges ed c i ical alues a
> 1
> 15
> 0.24
a No haizan e al., 2009.
Essen ial mine als especially i on and zinc a e among
he mos de icien mic onu ien s in A ica and a e
associa ed wi h poo g ow h and de elopmen and
impai ed immune esponse (WHO, 2017).
The de iciency o essen ial mine als in he human
body may be a esul o inadequa e in ake o impai ed
abso p ion o mine als due o he in e ac ion be ween
essen ial nu ien s o he p esence o phy a e and
oxala e in he die (No haizan and Faizada ul, 2009;
Nguyen e al., 2012).
I is o en ci ed ha a highe concen a ion o i on
which is abo e 2:1 mola a io can nega i ely a ec
zinc abso p ion especially when hey a e aken as a
solu ion (Nguyen e al., 2012; Rossande -Hul e al.,
2018). Fu he mo e, adequa e in ake o calcium, as
well as phospho us in he app op ia e a io (1-2:1), is
i al o bone heal h and in an de elopmen
(Lough ill e al.,2017).
Table 4. Concen a ions o o al phenolic and o al la onoid in Spi ulina pla ensis.
Nu ien
Concen a ion
To al phenolic (mg GAE/g)
2.99
To al la onoid (mg QE/g)
1.92
The compu ed mola a ios in his s udy
demons a ed ha he e is no impai men o nu ien
abso p ion due o in e ac ions be ween mine als o
be ween mine als and phy a e. Gu ié ez-Salmeán e
al. (2015) also epo ed no inhibi ion o spi ulina
nu ien abso p ion due o he p esence o
an inu i ional ac o s. Fu he mo e, due o he lack
o cellulose in spi ulina cell wall, spi ulina ends o be
easily diges ed by he human diges i e sys em (Dillon
and Phucb, 1995; Ka kos e al., 2011). In his s udy,
he sa e y o spi ulina o human consump ion was
e alua ed based on hea y me als concen a ion. Ou
126
Teze a e al.
In . J. Biosci.
2020
indings e ealed ha Spi ulina pla ensis has a e y
li le amoun o me cu y, lead, cadmium, and a senic
as compa ed wi h he ecommended maximum le el
o hea y me als in oods u se by he Eu opean
Commission (Eu open Commission Regula ion (EC),
2016). Mo eo e , s udies epo ed ha spi ulina has
nei he an acu e no ch onic oxicological e ec on
he human body (Naidu e al., 2009; Gu ié ez-
Salmeán e al., 2015; Neumann e al., 2018). Besides,
spi ulina is among he oods which a e gene ally
ecognized as sa e by he US Food and D ug
Adminis a ion (FDA, 2011).
Table 5. The concen a ion o hea y me als in Spi ulina pla ensis and he sugges ed maximum le el o hea y
me als in oods u (mg/kg).
Me al
Sample (mg/kg)
a Maximum le el (mg/kg)
Me cu y
0.000036
0.1
Lead
0.0047
0.1
Cadmium
0.00048
0.05
A senic
0.0047
0.1
a Eu opean Communi ies Commission Regula ion (EC) No 1881/2006 o 19 Decembe 2006. Se ing maximum
le els o ce ain con aminan s in oods u .
Conclusion
In ligh o i s nu i ional po en ial, we hus concluded
ha spi ulina pla ensis cul i a ed in Eas A ica is a
sa e sou ce o essen ial nu ien s. The e o e, i s
p oduc ion, as well as u iliza ion, should be highly
p omo ed as a sus ainable sou ce o al e na i e ood
supplemen s o add ess unde nu i ion in he egion.
Acknowledgmen
The au ho s would like o hank Cen e o Resea ch,
ag icul u al Ad ancemen , Teaching Excellence and
Sus ainabili y in Food and Nu i ion Secu i y
(CREATES- FNS) and A usha Technical College o
hei suppo .
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