Scientific African 19 (2023) e01548

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Assessment of nutritional and medicinal properties of nutmeg

Pius A. Okiki a,∗, Chizobam P. Nwobi a, Oghenerobor B. Akpor a, Ezekiel Adewole b,
Richard D. Agbana c
a
Department of Biological Sciences, Afe Babalola University, Ado-Ekiti, Nigeria
b
Department of Chemical Sciences, Afe Babalola University, Ado-Ekiti, Nigeria
c
Department of Community Medicine, Afe Babalola University, Ado-Ekiti, Nigeria

a r t i c l e i n f o a b s t r a c t

Article history: The study was aimed at assessing the nutritional and medicinal properties of nutmeg, a
Received 2 January 2022 seed of Myristica fragrans Houtt. Locally sourced nutmeg in Ado-Ekiti, Nigeria, were anal-
Revised 2 January 2023
ysed for their proximate, minerals, vitamins, phytochemicals and bioactive molecules con-
Accepted 9 January 2023
tents. GC–MS analysis of the essential oil was carried out and drug properties of the iden-
tified bioactive molecules were screened computationally, using online Osiris server. The
Editor: DR B Gyampoh methanolic and ethanolic extracts of nutmeg were screened for their antibacterial activi-
ties. Results: The nutmeg samples were found to have high contents of fat (26.7%), pro-
Keywords:
tein (18.7%), carbohydrate (28.9%), energy (3938.3 Kcal/Kg) and fibre (9.4%); rich in Ca²+ ,
Antibacterial profiles
K+ , Po4 3 − , Mg²+ , Fe²+ and ascorbic acid; with low levels of Zn2+ , thiamine, niacin and
Drug properties
Myristica fragran riboflavin. The phytochemical analysis indicated high levels of alkaloids, phytates, tan-
Nutmeg nins, oxalates, flavonoids, terpernoids, antioxidant, with low protease inhibitor, and no
Nutritional values detectable cyanogenic glycosides. The GC–MS analysis of the essential oil gave a chro-
matogram with 33 peaks, and the identified molecules including gamma-Terpinene, Isobor-
neol, Safrole, Geraniol, (+)-4-Carene, Myristicin and Guaiol among others, and some of
these molecules are having excellent drug properties. The multi-drug resistant clinical bac-
terial isolates and the reference bacteria used in this study were all susceptible to both
methanolic and ethanolic extracts of nutmeg, with the methanolic extract showing more
significant antibacterial profiles. The present study demonstrated nutmeg as having high
nutritional and antibacterial values, with rich content of bioactive molecules that could
serve as sources of new therapeutics against drug resistant bacterial infections.
© 2023 The Author(s). Published by Elsevier B.V. on behalf of African Institute of
Mathematical Sciences / Next Einstein Initiative.
This is an open access article under the CC BY license
([Link]

Introduction

Myristica fragrans belongs to the family Myristicaceae and is a tropical evergreen tree. It produces two spices, nutmeg
and mace; the most important part of the plant in terms of its pharmacological activity and also in commerce is the dried
kernel (seed), the nutmeg. The spice nutmeg has a distinctive fragrance and a slightly sweet taste. It is used globally, as a

∗
Corresponding author.
E-mail address: okikipa@[Link] (P.A. Okiki).

[Link]
2468-2276/© 2023 The Author(s). Published by Elsevier B.V. on behalf of African Institute of Mathematical Sciences / Next Einstein Initiative. This is an
open access article under the CC BY license ([Link]
P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

condiment to flavour baked foods, confections, sausages, sauces, meats, and in many other foods. This spice has been used
extensively in traditional medicine in treating several ailments [1–3].
Commercial antimicrobial drugs have been commonly employed for treatment of infectious diseases for many years.
However, the indiscriminate use of these antibiotics has developed multiple resistances and side effects [4]. Thus, more
natural antimicrobial substances from plants are desired. A large number of herbs possess antimicrobial activity [5] and
their active components have become a potential source of new antimicrobial agents [6,7]. Plant extracts have been studied
against bacteria, fungi and yeast; and spices and herbs used today are valued for their antimicrobial activities and medic-
inal effects in addition to their flavour and fragrance qualities. The antimicrobial activities of plant extracts form the basis
for many applications, including raw and processed food preservation, pharmaceuticals, alternative medicines and natural
therapies [8,9]
Nutmeg is used as a constituent in preparations of medicines, such as for dysentery, flatulence, stomach-ache, nausea,
vomiting, rheumatism, sciatica, malaria and early stages of leprosy. M. fragrans has been deductively approved to treat hy-
polipidemic and hypocholesterolaemia, antidepressant, aphrodisiac, antimicrobial, upper antioxidant, memory boosting, and
hepatoprotective properties. The essential oil of nutmeg and its fractions have been reported to contain pharmacological ac-
tive compounds with aphrodisiac, hepatoprotective, antimicrobial, antidiabetic, antioxidant, anticancer properties [2,3]. The
study was aimed at assessing the nutritional biomolecules in essential oil and antibacterial potentials of locally sourced M.
fragrans seeds.

Materials and methods

Plant sample collection: High-quality dried nutmeg, without defects, used in this research work were purchased from a
local market in Ado-Ekiti, Ekiti State, Nigeria (Plate 1).
Proximate, vitamins and minerals analyses of nutmeg: The proximate, vitamins and minerals were analysed using stan-
dard techniques prescribed by Association of Official Analytical Chemists [10]. For the proximate, the pulverized seeds of M.
fragrans were analysed for Moisture, fat, carbohydrate, crude fibre and protein contents. The minerals analysed were Ca²+ ,
K− , PO4 3 − , Mg²+ , Fe²+ and Zn2+ ; while the vitamins were ascorbic acid, thiamine, niacin and riboflavin.
Proximate: The seeds (nutmeg) were opened using a sharp stainless-steel knife, stripped from the shells and pulverized.
Moisture, protein, fat, and ash contents in the pulverized nutmeg samples were determined in accordance with the stan-
dard methods of the Association of Official Analytical Chemists (AOAC) procedures [10]. Moisture content was determined
gravimetrically by drying the samples in an oven at 100 °C to a constant weight. The percentage of moisture content was
calculated as follows:
(Initial weight of sample − final weight of sample) × 100
Moisture (% ) =
Initial weight of sample
Ash content was assayed by incinerating the samples in a muffle furnace at 550 °C (AOAC Method No. 930.05) [10]. The
remaining inorganic material was cooled in a desiccator, weighed and the ash content was determined as follows:
Weight of sample remaining × 100
Ash content (% ) =
Weight of original sample
The dried pulverized nutmeg was subjected to other chemical analyses. Crude protein content (N × 6.25) was deter-
mined in accordance with the Kjeldahl method (Method No. 978.04) [10]. The Kjeldahl method follows three steps namely
digestion, neutralization and titration. The nitrogen content was estimated by titration of the ammonium borate formed
with standard hydrochloric acid, using a suitable indicator to determine the end-point of the reaction. The following equa-
tion can be used to determine the nitrogen concentration of a sample that weighs m grams using a xM HCl acid solution
for the titration:
xmoles (vs − vδ )cm3 14g
%N = × × × 100
10 0 0cm3 mg moles
Where vs and vb are the titration volumes of the sample and blank, and 14 g is the molecular weight of nitrogen N.
Once the nitrogen content has been determined it is converted to a protein content using the appropriate conversion factor
of 6.25 (equivalent to 0.16 g nitrogen per gram of protein)
Crude fat was determined in accordance with the Soxhlet extract method using petroleum ether as the extract agent
(60–80 °C) (AOAC Method No. 930.09) [10]. The extracted fat concentrated using a rotary evaporator followed by drying in
water-bath at 45 °C. The crude fat concentration was determined as follows:
Weight of extracted fat×100
Fat (% ) =
Weight of original sample
The percentage carbohydrate (by difference) was calculated as 100 – (% moisture +% fibre +% protein +% fat)
Minerals: The pulverized nutmeg samples were digested by concentrated nitric acid and perchloric acid (4:1, v/v) at
70–90 °C for 10 min and then cooled for injection. The concentrations of the metals were determined by using an Atomic
Absorption Spectrophotometer (Buck Scientific model 210 VGP) [10]. Wavelengths, slits and lamp current used for the de-
termination of six elements were 213.9 nm, 0.5 nm, 4.0 mA (zinc); 422.7 nm, 1.2 nm, 4.0 mA (calcium); 248.3 m, 0.2 nm,

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

6.0 mA (iron) and 766.5 nm, 0.8 nm, 4.0 mA (potassium), respectively. The results for mineral contents were expressed as
mg/100 g DW. The total phosphate content in nutmeg samples was determined by spectrophotometric method, based on
the formation of phosphomolybdate complex with the added molybdate followed by the reduction of the complex with
thiourea in aqueous sulfuric acid medium.
Vitamins analyses: The vitamin B group was extracted according to a previously described method by Sami et al. [11] and
AOAC [10]. In brief, nutmeg powder (2 g) was placed in 25 mL of H2 SO4 (0.1 N) solution and incubated for 30 min at 121 °C.
Then, the contents were cooled and adjusted to pH 4.5 with 2.5 M sodium acetate, and 50 mg Takadiastase enzyme was
added. The preparation was stored at 35 °C overnight. The mixture was then filtered through a Whatman No. 4 filter, and
the filtrate was diluted with 50 mL of pure water and filtered again through a micropore filter (0.45 μm). a volume of 20
μL of the filtrate was injected into the HPLC system. Quantification of vitamin B content was accomplished by comparison
to vitamin B standards, using standard stock solutions for thiamine, riboflavin and niacin prepared as described by AOAC
[10]. Chromatographic separation was achieved on a reversed phase- (RP-) HPLC column (Agilent ZORBAX Eclipse Plus C18;
250 × 4.6 mm i.d., 5 μm) through the isocratic delivery mobile phase (A/B 33/67; A: MeOH, B: 0.023 M H3 PO4 , pH = 3.54)
at a flow rate of 0.5 mL/min. Ultraviolet (UV) absorbance was recorded at 270 nm at room temperature.
Vitamin C was extracted and analysed as previously described by Sami et al. [11]. The nutmeg powder (10 g) was blended
and homogenized with an extracting solution containing metaphosphoric acid (0.3 M) and acetic acid (1.4 M). The mixture
was placed in a conical flask and agitated at 10,0 0 0 rpm for 15 min. The mixture was then filtered through a Whatman No. 4
filter, and samples were extracted in triplicate. The ascorbic acid standard was prepared by dissolving 100 mg of l-ascorbic
acid in a metaphosphoric acid (0.3 M)/acetic acid (1.4 M) solution at a final concentration of 0.1 mg/mL. The calibration line
was converted to a linear range based on four measured concentration levels. Quantification of ascorbic acid content was
performed on an Agilent HPLC system. Chromatographic separation was achieved on an RP-HPLC column through isocratic
delivery of a mobile phase (A/B 33/67; A: 0.1 M potassium acetate, pH = 4.9, B: acetonitrile: water (50:50) at a flow rate of
1 mL/min. UV absorbance was recorded at 254 nm at room temperature.
Quantitative phytochemical analysis: Quantification of phytochemicals: alkaloids, flavonoids, steroids, saponins, phenols,
tannins, glycosides, cardiac glycosides phytates, total phenol, protease inhibitors and antioxidants were carried out on the
powdered nutmeg samples following methods described by Harbone [12].
Determination of alkaloids: This was done by adding to 5 g of the powdered nutmeg, 200 mL of 10% acetic acid in ethanol
and allowed to stand for 4 h. It was filtered and the filtrate was concentrated on a water bath to one quarter of the original
volume; then Conc. NH4 OH added drop wise to the filtrate until complete precipitation was obtained. The precipitate was
collected and washed with dilute NH4 OH and filtered. The residue is the alkaloid, which was dried and weighed.
Estimation of Flavonoids concentration: This was carried out as follows: 10 g of powdered nutmeg was extracted with
100 mL of 80% aqueous methanol at room temperature, and the extract filtered with the aid of Whatman filter paper. The
filtrate was transferred into a water bath and allowed to evaporate to dryness and weighed.
Estimation of total Saponins: This was done by adding to 20 g of the ground sample of nutmeg 100 mL of 20% aqueous
ethanol. The extract was concentrated and purification process repeated the purification process was repeated with 20%
aqueous ethanol, followed by addition of n-butanol. The combined n-butanol extracts were washed twice with 5% aqueous
sodium chloride. The remaining solution was heated in a water bath for evaporation and was further dried in the oven to a
constant weight; the saponin.
Antioxidant activity: This was measured using (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity assay. The scav-
enging ability of the different plant fractions and standard was calculated using the equation:
(Absorbance o f control − Absorbance o f sample) × 100
Scavengingact ivit y (% ) =
Absorbance o f control
Preparation of nutmeg extracts: One hundred gram (100 g) dry M. fragrans seed powder each was soaked separately in
500 mL of methanol, 500 mL of ethanol and 500 mL of N–Hexane for 72 h at room temperature and filtered with Whatman
No 1 filter papers. The filtrates were condensed, using a rotary evaporator and the residues were dried up with the aid of
water bath at 40 °C, and refrigerated at 4 °C till used. The ethanol and methanol extracts were used for investigation of
antimicrobial potential of nutmeg, while the N–Hexane extract was used to identify biomolecules content of the plant that
are of health benefit [12].
GC–MS analysis of drug properties of Myristica fragrans oil: Identification of the chemical compounds present in the
N–Hexane extract of nutmeg was carried out by the Gas Chromatography Mass Spectrometry (GC–MS). GC–MS analysis was
carried out on a GC–MS-QP 2010 Plus Shimadzu system and Gas chromatograph interfaced to a mass spectrometer (GC–MS)
instrument employing the following conditions: Column Elite-1 fused silica capillary column (30 m x 0.25 mm 1D x μL df,
composed of 100% dimethyl polysiloxane). For GC–MS detection, an electron ionization system with ionization energy of
70 eV was used. Helium gas was used as the carrier gas at constant flow rate 1 mL/min and an injection volume of 2μL was
employed (Split ratio of 10:1) injector temperature of 250 °C; ion-source temperature 280 °C. The oven temperature was
programmed from 110 °C (Isothermal for 2 min) with an increase of 10 °C / min to 200 °C then 5 °C/min to 280 °C/min,
ending with a 9 min. isothermal at 280 °C. Mass spectra were taken at 70 eV; a scan interval of 0.5 s and fragments from
40 to 550 Da. Total GC running time was 36 min. The relative percentage amount of each component was calculated, by
comparing its average peak area to the total areas, Software adopted to handle mass spectra and chromatogram was a
turbomass. The detection employed the NIST Ver. 2.0 year 2009 library [13].

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

Table 1
Nutritional compositions of nutmeg.

Analyte Concentration

Proximate
Moisture Content (%) 9.37 ± 0.06
Protein (%) 18.67 ± 0.21
Ether Extract/Fat (%) 26.73 ± 0.25
Ash (%) 3.87 ± 0.21
Crude Fibre (%) 12.43 ± 0.12
Carbohydrate (%) 28.93 ± 0.61
Energy (Kcal/kg) 3938.33 ± 2.08
Minerals
Fe+ + (mg/100 g) 7.63 ± 0.15
Zn+ + (mg/100 g) 0.53 ± 0.06
Mg+ + (mg/100 g) 56.67 ± 2.98
Ca+ + (mg/100 g) 166.67 ± 7.64
K+ (mg/100 g) 68.33 ± 2.89
PO4 − (mg/100 g) 141.67 ± 7.64
Vitamins
Ascorbic acid (mg/100 g) 14.77 ± 0.25
Thiamine(mg/100 g) 0.10 ± 0.01
Niacin(mg/100 g) 1.2867 ± 0.02
Riboflavin(mg/100 g) 0.10 ± 0.01

Values are expressed in mean ± SD.

Some of the GC–MS identified compounds in the crude extracts of M. fragrans were screened computationally for pos-
session of drug properties, using online OSIRIS property explorer server. Each of the compounds were assessed for their
bioavailability, hydrophobicity and membrane permeability which are linked with some molecular descriptors such as cLog
P (partition coefficient) and cLogS (solubility) [14].
Collection of bacterial culture: Ten clinical and three typed bacterial isolates were obtained from the bacterial collections
of Microbiology Laboratory, Afe Babalola University, Ado-Ekiti, Nigeria. The bacteria used in this study were, Klebsiella pneu-
moniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Enterobacter aerogenes, Acinectobacter baumanii, Staphylo-
coccus aureus, Corynebacterium accolens, Enterococcus faecalis, Propionibacterium acnes, Staphylococcus aureus (ATCC 25923),
E. coli (ATCC 35218) and Proteus mirabilis (ATCC 12453). The bacteria were sub-cultured on freshly prepared nutrient agar
plates at 37 °C for 24 h before use.
Antibiotic susceptibility test: The clinical bacterial isolates were tested for antibiotics susceptibility by Kirby-Bauer
disc diffusion method on Mueller-Hinton agar (Clinical and Laboratory Standards Institute [15]. Antibiotics used were:
Trimethoprim-Sulfamethoxazole SXT (30 μg), Chloramphenicol C (30 μg), Sparfloxacin SPX (10 μg), Ciprofloxacin CIP (10 μg),
Amoxicillin AMX (30 μg), Amoxycillin-Clavulanic acid AUG (30 μg), Gentamycin GEN (10 μg), Pefloxacin PEF (30 μg),
Ofloxacin OFL (10 μg), Streptomycin STR (30 μg) for gram negative isolates; while Pefloxacin PEF (10 μg), Gentamycin GEN
(10 μg), Ampicillin-Cloxacillin AMP (30 μg), Cefuroxime CXM (30 μg), Amoxicillin AMX (30 μg), Ceftriaxone CRO (30 μg),
Ciprofloxacin CIP (10 μg), Streptomycin STR (30 μg), Amoxycillin-Clavulanic acid AUG (30 μg) and Erythromycin ERY (10 μg),
for gram positive isolates.
Antimicrobial activity of M. fragrans seed extracts using agar-well diffusion method: Susceptibility of the test organisms
to ethanolic and methanolic extracts of nutmeg was determined using the agar well diffusion method on Mueller-Hinton
agar. Seven-millimetre diameter wells were prepared on agar containing a suspension of each isolate. The respective extracts
were diluted into two folds and known concentrations were placed in the wells. The plates were left at ambient temperature
for about 15 min and then incubated at 37 °C for 24 h, after which zones of inhibition recorded were by subtracting the
well diameter [9,16].
Statistical analysis of data: Data were expressed as mean ± standard deviation. One Way ANOVA (using IBM-SPSS Statis-
tics 23) was used to test significant differences between bacterial isolates susceptibility to ethanolic and methanolic extracts
of nutmeg, with values of p ≤ 0.05 considered to be significant.

Results

Chemical analysis of nutmeg seeds revealed high nutritional and phytochemical contents, as well as an array of bioactive
molecules in the essential oil.
Nutritional content of nutmeg: The samples of nutmeg were found to be high in fat (26.73 ± 0.25%), protein
(18.67 ± 0.21%), carbohydrate (28.93 ± 0.61%), energy (3938.33 ± 2.08 KCal/Kg) and fibre (12.43 ± 0.12%); rich in Ca²+ ,
K− , PO4 ²− , Mg²+ , Fe²+ and ascorbic acid; but with low levels of Zn2+ , thiamine, niacin and riboflavin (Table 1).
Phytochemical contents: The quantitative phytochemical analysis indicated high level of alkaloids
(858.33 ± 17.56 mg/100 g), phytates (390.00 ± 5.00 mg/100 g), tannins (1248.33 ± 16.07 mg/100 g), oxalates
(225.0 0 ± 5.0 0 mg/10 0 g), flavonoids (558.33 ± 16.07 mg/100 g) and terpernoids (1538.33 ± 12.58 mg/100 g), an-

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

Table 2
Phytochemical compositions of nutmeg.

Analyte Concentration

Alkaloids (mg/100 g) 858.33 ± 17.56

Phytates (mg/100 g) 390.00 ± 5.00
Tannins (mg/100 g) 1248.33 ± 16.07
Saponins (mg/100 g) 23.33 ± 7.64
Oxalates (mf/100 g) 225.00 ± 5.00
Flavonoids (mg/100 g) 558.33 ± 16.07
Terpenoids (mg/100 g) 1538.33 ± 12.58
Cyanogenic glycosides (mg/100 g) ND
Total phenol (GAE/g) 52.47 ± 0.21
Protease inhibitor (mg/100 g) 0.05 ± 0.02
DPPH Scavenging activity (%) 31.27 ± 0.25

Values are expressed in mean ± SD.

ND – Not detected.

Fig. 1. GC–MS spectra of nutmeg oil.

tioxidant (31.50 ± 0.21%), very low protease inhibitor (0.05 ± 0.02 mg/100 g) with no detectable cyanogenic glycosides
(Table 2).
Bioactive molecules: The GC–MS analysis of the essential oil gave spectra revealed 33 peaks (Fig. 1). Biomolecules iden-
tified were gamma-Terpinene, Isoborneol, 2-methylisoborneol, Safrole, Geraniol, (+)−4-Carene, Myristicin and Guaiol among
others (Table 3). Drug properties of some of the identified compounds as determined by OSIRIS property explorer are pre-
sented in Table 4. Excellent drug properties were obtained for 2-methylisoborneol, Terpineol, Geraniol and Guaiol, and they
were found to be non-mutagenic and non-tumorigenic.
Antibacterial profiles of nutmeg extracts: The ten clinical bacterial isolates used in the study were resistant to multiple
drugs (Fig. 2, Table 5). All the clinical bacterial isolates and the three reference bacteria were susceptible to both ethanolic
and methanolic extracts of the nutmeg, showing similar patterns of susceptibility. However, the methanolic extract produced
significantly higher susceptibilities than the ethanolic extracts with respect to Corynebacterium accolens (p = 0.006), Propi-
onibacterium acnes (p = 0.002), Staphylococcus aureus ATCC 25923 (p = 0.005) and Eschericia coli ATCC 35218 (p = 0.016),
(Table 6).

Discussion

The nutmeg investigated in this study was found to contain high levels of fat, protein and carbohydrate, which con-
tribute to the high level of energy. Carbohydrates in the human body serve various functions which include; energy produc-

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

Table 3
Identified compounds from GC–MS spectra of nutmeg oil.

Peak no Retention time (min) % Area (quality) Compound name

1 5.626 0.97 (+)−4-Carene

2 6.210 1.62 .gamma.-Terpinene
3 6.328 1.56 2-Cyclohexen-1-ol
4 6.758 1.85 Linalol
5 .028 2.44 trans-4–methoxy thujane
6 7.682 0.63 Isoborneol
7 7.849 2.21 Terpinen-4-ol
8 8.000 1.15 [Link].-Terpineol
9 8.047 2.21 cis-4–methoxy thujane
10 8.786 0.60 Geraniol
11 9.222 8.25 Safrole
12 9.898 0.31 alpha.-Cubebene
13 10.230 1.93 Copaene
14 10.472 3.90 Methyleugenol
15 10.746 2.66 Caryophyllene
16 10.841 0.61 Bergamotene
17 10.953 0.16 Alloaromadendrene
18 11.108 0.48 Humulene
19 11.312 0.47 .gamma.-Muurolene
20 11.390 0.20 1,6-Cyclodecadiene
21 11.546 1.59 Aromandendrene
22 11.607 0.60 beta.-Bisabolene
23 11.717 0.20 .gamma.-Muurolene
24 11.903 27.43 Myristicin
25 12.159 19.15 Asarone
26 12.448 0.35 [Link].-Bisabolene epoxide
27 12.602 0.62 Guaiol
28 13.024 2.92 Isoelemicin
29 13.151 0.92 2-Naphthalenemethanol
30 3.275 0.41 5-Azulenemethanol
31 16.344 0.20 1H–Naphtho[2,1-b]pyran
32 17.517 0.45 trans-Geranylgeraniol
33 18.341 0.52 Longifolene

Table 4
Drug properties of some of identified compounds in the crude extracts of nutmeg determined by OSIRIS property explorer.

Compound Drug likeness Mutagenic Tumorigenic Reproductive effect Irritability clog P clog S
a
3-Carene −3.3517 None High None High 2.721 −2.519
b
2-methylisoborneol −1.6505 None None None None 2.3336 −2.51
c
Terpineol −3.3493 None None None Low 2.2993 −2.186
d
Geraniol −3.5697 None None None High 3.4853 −1.889
e
Guaiol −2.0249 None None None None 3.6689 −3.256
f
Myristicin −4.6675 High High None None 2.7294 −3.057
g
Asarone −4.0375 High High High None 2.5402 −2.578
Longifolene −7.7594 None None None None 4.0602 −3.809
Bergamotene −2.3633 None None None High 4.8928 −3.495
a
3-Carene shares the same functional group with (+)−4-Carene.
b
2-Methylisoborneol shares the same functional group with Isoborneol Bicyclo[2.2.1] heptan-2-ol.
c
Terpineol shares the same functional group with alpha.-Terpineol.
d
Geraniol shares the same functional group with Geraniol.
e
Guaiol shares the same functional group with Guaiol 5-.
f
Myristicin shares the same functional group: methoxy– Myristicin.

tion, energy storage, building macromolecules, sparing protein, and assisting in lipid metabolism among others. Proteins are
molecules made of amino acids and play a central role in biological processes. For example, proteins catalyse reactions in
our bodies, transport molecules such as oxygen, keep the body healthy as part of the immune system and transmit messages
from cell to cell [17]. The result proximate compositions of nutmeg in present study was similar to those reported by Asika
et al. [18] and that of Bamigboye et al. [19]. The present study, however, reported higher protein concentration than those
reported by the other two workers, which might be due to low moisture content of 9.37 in this study compared to 14.2 and
28.8 reported by Asika et al. and Bamigboye et al. respectively.
For the mineral and vitamin contents, the nutmeg seeds were found to be rich in Ca²+ , K− , PO4 ²− , Mg²+ , Fe²+ and ascor-
bic acid, with low levels of Zn²+ , thiamine, niacin and riboflavin (Table 1). Vitamins and minerals are considered essential
nutrient, because acting in concert, they perform hundreds of roles in the body. They help shore up bones, heal wounds,
and bolster your immune system. They also convert food into energy, and repair cellular damage. Potassium is an important

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

Fig. 2. Antibiotics susceptibility patterns of clinical bacterial isolates.

Plate 1. Photograph of nutmeg seeds.

Table 5
Cluster of antibiotics resistance pattern of used clinical bacterial isolates.

Codes Organisms Cluster of resistance

3c Propionibacterium acnes PEF/AMP/CXM/AMX/CRO/ERY

8c Acinectobacter baumanii PEF/AMP/CXM/AMX
14c Enterococcus faecalis PEF/AMP/CXM/AMX/CRO/CIP
17a Corynebacterium accolens PEF/GEN/AMP/CXM/AMX/CIP/ERY
4d Staphylococcus aureus PEF/AMP/CXM/AMX/CRO/AUG
134d Klebsiella pneumoniae SXT/C/SPX/AMX/AUG/GEN/PEF/OFX/STR
44a Enterobacter aerogenes AMX/AUG/GEN/PEF
23e Pseudomonas aeruginosa SXT/C/SPX/CIP/AMX/AUG/GEN/PEF/OFX/STR
7b Proteus mirabilis AMX/AUG/OFX/STR
12d E. coli C/SPX/AMX/AUG/PEF/OFX/STR

Trimethoprim-Sulfamethoxazole (SXT), Chloramphenicol (C), Sparfloxacin (SPX),

Ciprofloxacin (CIP), Amoxicillin (AMX), Amoxycillin-Clavulanic acid (AUG), Gen-
tamycin (GEN), Pefloxacin (PEF), Ofloxacin (OFL), Streptomycin (STR), Pefloxacin (PEF),
Ampicillin-Cloxacillin (AMP), Cefuroxime (CXM), Ceftriaxone (CRO), Streptomycin (STR)
(30 μg) and Erythromycin (ERY).

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Table 6
Inhibitory effects of the methanolic and ethanolic extracts of nutmeg against test organisms.

Acineto- Entero- Pseudo- Staphylo- Proteus E. coli

Concentra- Klebsiella Proteus Corynebact- Staphyloco- Propionibact- Esheri- bacter Enteroco- bacter monas coccus aureus mirabilis (ATCC
Extracts tions (mg/mL) kneumoniae mirabilis erium accolens ccus aureus erium acnes chia coli baumani ccus faecalis aerogenes aeruginosa (ATCC 25923) ATCC 12453 35218)

Methanolic 100 14 15 17 10 14 11 18 19 20 15 12 12 7
50 13 13 15 8 12 9 16 17 18 13 10 10 5
25 11 11 13 5.5 10 7 14 15 15 12 8 8 3
12.5 10 8 11 4.5 8.5 5 12 13 15 12 6 10 2
6.25 9 8 0 7 5.5 3 10 11 7 8 5 6 2
3.13 6 7 5 3 4 6 8 12 8 8 4 3 2.5
1.56 5 3 5 2.5 3 5 0 8 13 5 3 4 2
0.78 3 4 3 2 2.5 5 0 3 8 4 4 3 2
0.39 0 0 2 0 0 0 0 0 4 0 0 0 0
8

Ethanolic 100 23 12 0 11 17 14 17 19 22 11 14 9 10
50 21 10 0 9 15 12 15 17 20 11 12 7 8
25 19 8 0 7 13 10 13 15 18 9 10 5 6
12.5 17 9 0 6 8 8 11 13 8 10 8 7 4
6.25 15 8 0 7 8 8 8 11 13 8 4 7 5
3.13 6 3 0 3 8 13 7 10 14 14 6 8 7
1.56 0 3 0 3 9 6 13 13 7 7 5 5 3
0.78 0 0 0 0 5 5 5 8 8 8 5 4 0
0.39 0 0 0 0 0 0 0 4 4 0 0 0 0
∗∗
P values 0.230 0.334 0.006∗ 0.317 0.002∗ 0.005∗ 0.477 0.306 0.679 0.924 0.005∗ 0.649 0.016∗

Zones of inhibition of varied concentrations of extracts of nutmeg in mm.

∗
Significant.
∗∗
P values of comparison of the zones of inhibition along column between methanolic and ethanolic extracts.

Scientific African 19 (2023) e01548

P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

component of cell and body fluids that helps control heart rate and blood pressure. It also plays a role in many body func-
tions including transmission of nerve signals, muscle contractions, fluid balance, and various chemical reactions. Iron is best
known for ferrying oxygen throughout the body, Zinc helps blood clot and it is essential for taste and smell, and bolsters
the immune response, while calcium, phosphorus and magnesium are important in maintaining quality bones [20]. It also
contains many vital B-complex vitamins, including folic acid, riboflavin and niacin; vitamin C, vitamin A and many flavonoid
anti-oxidants like beta-carotene and cryptoxanthin that are essential for optimum health (20). Okonkwo and Ogu [21] had
earlier reported high concentrations of various vitamins and minerals in M. fragrans.
The present study demonstrated high levels of alkaloids, phytates, tannins, oxalates, flavonoids and terpenoids with low
levels of saponins, antioxidant and protease inhibitor in powdered nutmeg. The phytochemical contents of nutmeg reported
in this study were similar to those reported by Asika et al. [18] and Bamigboye et al. [19]. Phytochemicals found in plant
sources have been reported to exert functions, such as antimicrobial, anti-cancer, antioxidant, anti-diabetic, hypolipidaemic
and hypocholesterolamic activiteses among others [22–24].
The GC–MS analysis of the essential oil gave a chromatogram showing biomolecules, such as Myristicin, gamma-
Terpinene, Isoborneol, Safrole, Geraniol, (+)−4-Carene, Longifolene, isoelemicin among others. M. fragrans fruits have been
reported to contain alkyl benzene derivatives including myristicin, elemicin and safrole, terpenes, alpha-pinene, myristic
acid, trimyristicin [25]. These active chemicals in M. fragrans seeds have many therapeutic applications in many traditional
medicines such as anti-fungal, anti-depressant, aphrodisiac, digestive, and carminative functions. Safrole is a precursor of
many products used as natural insecticides as well as in the perfumery industries. Studies have shown that safrole pos-
sessed antioxidant and antimicrobial activities [26]. Geraniol has been found to possess antimicrobial, ani-inflammatory
and antioxidant activities [27]. Humulene is an immunomodulator and anti-inflammatory [28]. Caryophyllene is a potent
anti-inflammatory agent with cytoprotective effects on gastric mucosa [29]. Campbell and co-workers [30] demonstrated
moderate antimalarial effect of Caryophyllene against two strains of Plasmodium falciparum by an essential oil rich in β -
caryophyllene and α -terpineol. Alloaromadendrene is a potent antioxidant. Essential oil alloaromadendrene from Cinnamo-
mum osmophloeum leaves has been used to prolong the life spans in Caenorhabditis elegans [31]. Cubebene has been reported
for its antimicrobial activities [32]. Carene, a bicyclic mono-terpenoid, has been reported by McPartland and Russo [33] as
a potent anti-inflammatory agent. Türkez et al. reported the ability of copaene, a tricyclic sesquiterpene, to increase the
antioxidant capacity in human lymphocyte cultures [34].
The distinctive odour of nutmeg is due to presence of essential oil which contains terpenes such γ -terpinene, terpene
derivatives (e.g. terpinol, geraniol, and linalool) and phenylpropanes (e.g. myristicin, safrole, and elemicin), were all re-
ported in this study. Nutmeg has been reported to show hallucinogenic effect due to the presence of hallucinogenic phenyl-
propanes, and these phenylpropanes are hepatotoxins and are considered being harmful for frequent users [35].
The characterized and the identified compounds exhibited various drug properties, such as drug likeness, toxicological
properties such as mutagenic, tumorigenic, irritability and reproductive effects. Nine compounds were screened for various
drug properties and all showed negative signs for drug likeness which is an indication that they are not pure drugs but
can be incorporated into the rings of compounds for the formulation of drug candidates. Methylisoborneol, Guaiol, Longifo-
lene and Geraniol have no toxicological effects. These compounds have earlier been found to have various pharmaceutical
applications.
The medicinal property of nutmeg was further demonstrated by antibacterial inhibitory effectiveness of methanolic and
ethanolic extracts of high-quality dried nutmeg. The clinical bacteria used in this study were resistant to multiple drugs.
Drug resistance among microorganisms have been found to be caused by overuse and misuse of drugs in humans and
animals [4,36]. The methanolic extract of nutmeg was effective against the typed bacteria and all the multidrug resistant
bacteria tested. Likewise, the ethanolic extract was effective against the type bacteria and the multidrug resistant bacteria
except the Corynebacterium accolens. The methanolic extract however produced significant higher antibacterial inhibition in
five of the bacteria than the ethanolic extract. These findings were in agreement with the report of previous study by Rani
and Khullar [21] that showed strong antibacterial activities of methanol extract of M. fragrans against multi-drug resistant
organisms.
In conclusion the present study demonstrated nutmeg was of high nutritional values and contain biomolecules of high
medicinal properties that can be harnessed for development of new therapeutics, especially against drug resistant bacterial
infections.

Funding

This work did not receive any external funding.

Declaration of Competing Interest

We the authors of the manuscript titled: Assessment of Nutritional and Medicinal Properties of Nutmeg, declare that
they have no known competing financial interests or personal relationships that could have appeared to influence the work
reported in this paper.

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P.A. Okiki, C.P. Nwobi, O.B. Akpor et al. Scientific African 19 (2023) e01548

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