Scientia Horticulturae 295 (2022) 110894

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Moringa leaf: An innovative source of antioxidative phenolics for

cosmeceutical products
Mayuree Kanlayavattanakul a, b, Nattaya Lourith a, b, *
a
School of Cosmetic Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
b
Phytocosmetics and Cosmeceuticals Research Group, Mae Fah Luang University, Chiang Rai 57100, Thailand

A R T I C L E I N F O A B S T R A C T

Keywords: Moringa oleifera Lam. (Moringaceae) or Moringa or miracle tree has been utilized in several forms in food and
Moringa nutraceuticals. However, preparation of the antioxidative, phenolic-rich fraction of Moringa leaves is sparsely
Miracle tree reported as per its status as an innovative product. A concise and processable extraction yielding phenolic-rich
Antioxidant
fractions derived from Moringa leaves was developed. Maceration of mature Moringa leaves in 70% ethanolic
Phenolic extract
Processing innovations
water for 24 h at room temperature afforded a significant phenolic extract with antioxidant activity (10.92 ±
0.23 g GAE/100 g extract and IC50 = 50.22 ± 0.09 µg/ml) significantly greater (p < 0.05) than those resulting
from other methods. The extract was applicable for topical products in terms of its physicochemical properties
and compatibility, resulting in a stable water-in-silicone (W/Si) emulsion. This antioxidative, phenolic-rich
Moringa emulsion can be used as a cosmeceutical product. The innovative Moringa product presented is
meeting the consumers’ preferences for natural/sustainable products that are continuing to increase year by
year. In addition, successive integration between the agricultural crop and cosmetic industries are evidenced and
significance to horticulture.

interest among researchers. In addition, consumers’ awareness and

1. Introduction
Moringa oleifera Lam. (Moringaceae), or Moringa or Drumstick, has
long been regarded as a miracle tree with several beneficial properties
(El-Serafy and El-Sheshtawy, 2020). Its pharmacological activities are
not only contributing to its therapeutic effects but also include positive
nutritional effects (Oyeyinka and Oyeyinka, 2018). This crop is there­
fore industrially cultivated worldwide and its safety for oral and topical
administration have been confirmed. The leaves, flowers, pods and seeds
of Moringa have been consumed to promote its health benefits. In
particular, Moringa leaves exhibit a wide range of biological activities
due to their health benefits derived from phenolics, i.e., cinnamic acid
derivatives including chlorogenic and caffeic acids, and quercetin, etc.
(Rodríguez-Pérez et al., 2015; Stohs and Hartman, 2015; Vongsak et al.,
2013). Phenolics are of great significance and importance for human
health and well-being (Kedrina-Okutan et al., 2019). They are therefore
utilized in several types of products, especially foods, nutraceuticals
(Meireles et al., 2020) and cosmeceutical products (Gunia-Krzyżak et al.,
2018; Zillich et al., 2015).
The current concept of beneficial agricultural extension in concert
with sustainable utilization of the available crops is gaining considerable
expectation for nutraceuticals, pharmaceuticals and cosmetics contain­
ing sustainable ingredients are increasing. These emerging trends
require the use of food crops in different industries in addition to their
conventional usages. The successive convergence of the food and
cosmetic industries is dramatically promoted, consistent with con­
sumers’ preferences for safe and efficient ingredient surpluses within the
biocircular economy and sustainable development of food crops (Far­
ia-Silva et al., 2020).
Despite the therapeutic effects of this miracle tree that have been
continuously revealed, preparation of the antioxidative, phenolic-rich
fraction of Moringa leaf to be supplied as the specialty material for
certain, value-added up industries, i.e., pharmaceuticals and cosmetics,
are sparsely reported, as well as the development of topical product
containing Moringa extract Ali et al., 2013; 2014). Moringa is therefore
highlighted as a ready-available plant to be developed as an alternative
health product in the form of cosmetics. The concise preparation method
yielding the phenolic-rich fractions derived from Moringa leaf fitting
with the industrial feasibility, was developed and comparatively
assessed for the antioxidant activities. Basic information on topical
product development, i.e., miscibility and pH, were examined. There­
after, a stable topical product containing the selected extract in

* Corresponding author.
E-mail address: nattayal@mfu.ac.th (N. Lourith).

https://doi.org/10.1016/j.scienta.2022.110894
Received 23 November 2021; Received in revised form 7 January 2022; Accepted 8 January 2022
Available online 13 January 2022
0304-4238/© 2022 Elsevier B.V. All rights reserved.
M. Kanlayavattanakul and N. Lourith Scientia Horticulturae 295 (2022) 110894

2018).
Abbreviations
2.3. Miscibility and pH evaluation
DI deionized
DPPH 1,1-Diphenyl-2-picrylhydrazyl Miscibility of the extract in the pharmaceutical solvents for topical
EtOH ethanol product, i.e., propylene glycol and glycerin were determined individu­
gGAE g of gallic acid equivalents ally including pH (Mettler Toledo, s20, Switzerland) (Kanlayavattana­
h hour kul et al., 2017).
HC. heat-cool
IC50 inhibitory concentration at 50% 2.4. Development of cosmetics containing the antioxidative, phenolic-rich
Int. initial moringa leaf extract
min minute
O/W oil-in-water The base W/Si emulsions containing the ingredients listed in Table 1
rpm rounds per minute were formulated, and pH and viscosity recorded (Brookfield DVII+ Pro,
TPC total phenolics content USA). Sensory evaluation onto the formulated preparation was moni­
W/O water-in-oil tored during the course of the product development by the formulator
W/Si water-in-silicone (Lourith and Kanlayavattanakul, 2017). Those that were preferred (data
not shown) were up-scaled and included for the physicochemical
property and stability assessments. Accelerated stability assessments
were undertaken by means of a centrifugation assay. Those that were
water-in-silicone (W/Si) emulsion was developed. Accordingly, a remained homogeneous were further included in the 2nd stability
bio-based product derived from the antioxidative, phenolic-rich Mor­ challenged condition, i.e., heat (45 ± 2 ◦ C, 24 h)-cool (4 ± 2 ◦ C, 24 h) for
inga leaf extract in the form of a W/Si emulsion for topical application is 7 cycles. The stable base formula was selected and incorporated with the
presented herein. selected Moringa extract and re-challenged under the same stability
assessment conditions (Kanlayavattanakul et al., 2017).
2. Materials and methods
2.5. Statistical analysis
2.1. Chemicals and reagents
Data are presented as the mean ± SD. The parameters were
The chemicals used for extraction were of commercial grade. Total compared (post hoc multiple comparison) and analyzed using ANOVA
phenolic content and antioxidant activity assays were of reagent grade. test with a significance level of p < 0.05 using the SPSS program version
Those for topical product formulation were of cosmetic grade. 16.0.

2.2. Preparation of antioxidative phenolic-rich moringa leaf extract 3. Results

Dried Moringa leaves were supplied by Thai traditional drug store
allocated in Bangkok. The plant material was identified by a botanist Dr.
Nijsiri Ruangrungsri, Faculty of Pharmaceutical Sciences, Department of
Pharmacognosy, Chulalongkorn University, Bangkok, Thailand. The
voucher specimen (MKMO 0401) was deposited for further reference at
our laboratory herbarium at Mae Fah Luang University, Chiang Rai. The
ground leaves were macerated in the solvents (1: 5, w/v) (Waterman
et al., 2014) that were deionized (DI) water, alcoholic water, i.e., 70%
EtOH, 60% EtOH and 50% EtOH at different time periods, with shaking
at 150 rpm, separately. The extracts were filtered (Whatman filter paper
no. 1) and concentrated to dryness under vacuum using either lyophi­
lizer or rotary evaporator, separately. The extraction under the same
condition was repeated twice and the extractive yield was calculated.
2.2.1. Total phenolics content (TPC)
TPC of each extract was determined using Folin-Ciocalteu assay. The
reagent was mixed with Na2CO3 and absorbance measured using the
microplate reader (ASYS, UVM340, UK). TPC was compared with gallic
acid and expressed as g of gallic acid equivalents (gGAE) per 100 g
extract. The procedure was repeated in triplicate (Kanlayavattanakul
et al., 2018).
2.2.2. DPPH● scavenging activity
Antioxidant activity was assessed with a DPPH assay. DPPH in ab­
solute EtOH reacted with standard ascorbic acid to generate a calibra­
tion curve (r > 0.999). Scavenging activity of a sample (150 µg/ml)
against DPPH● was monitored at 517 nm using a microplate reader. The
radical terminating capability (%) of the extract was calculated in
comparison with the standard values. The most potent antioxidant
extract was evaluated on its IC50 (inhibitory concentration at 50%). The
experiments were performed in triplicate (Kanlayavattanakul et al.,
3.1. Preparation of antioxidative, phenolic-rich moringa leaf extract
Moringa leaves with an optimized 1:5 ratio of plant material and
solvent (Waterman et al., 2014) were extracted with water at a different
time (15 min, 30 min, 1 h, 3 h and 6 h). That of 3 h was significantly (p <
0.05) greater than others (34.57 ± 1.79%, as depicted in Fig. 1.
Thereafter, ethanolic water was substantively used as the solvent (Sid­
dhuraju and Becker, 2003). Alcoholic water extractions with higher
water contents and longer extraction durations insignificantly increased
the extractive yields (p > 0.05), as shown in Fig. 1.
TPC, and antioxidant activity were screened at a concentration of
150 µg/ml. The 70% EtOH extract obtained after 24 h was significantly
(p < 0.05) richer in TPC (10.92 ± 0.23 g GAE/100 g), consistent with its
potent antioxidant activity (76.79 ± 0.59%). This extract was thus
selected and evaluated for its IC50. The antioxidant activity of this
Moringa leaf extract was weaker than that of the standard ascorbic acid
(IC50 = 50.22 ± 0.09 and 5.16 ± 0.71 µg/ml).
3.2. Development of a stable W/Si emulsion containing moringa leaf
extract
Basic information on miscibility and pH were first determined. The
extract was more miscible in propylene glycol than in glycerin (15.84 ±
1.45% and 3.33 ± 1.45%). The pH of the extract (5.55 ± 0.17 and 5.47
± 0.19) was suitable for its use as a topical product ingredient (Lourith
and Kanlayavattanakul, 2017).
A stable base W/Si emulsion formula containing the ingredients
listed in Table 1 was first developed. Propylene glycol, an appropriate
diluent for the extract, was incorporated into the designed formulation.
In addition, it functioned as a humectant in the preparation of glycerin.
Cetyl dimethicone, cyclomethicone and cyclopentasiloxane contributed

2
M. Kanlayavattanakul and N. Lourith Scientia Horticulturae 295 (2022) 110894

Table 1
Development of W/Si base emulsions and the W/Si emulsion containing Moringa leaf extract.
Ingredient Formula (% w/w)
A B C D E

Dimethicone 4 3.5 3 3 3
Lauryl PEG/PPG 18–18 methicone 17 17.5 17 18 18
Cetyl dimethicone
Cyclomethicone
Cyclopentasiloxane – – –
DI water
Glycerin 66.6 67.1 67.6 66.6 62.35
Propylene glycol
DI water
Sodium chloride 12.4 12.4 12.4 12.4 12.4
Germaben® II
Propylene glycol – – – – 3.75
Moringa leaf extract – – – – 0.5
Physicochemical property (Int.)
Viscosity (cPs) 15,895.00 ± 25.42 20,289.00 ± 67.00 13,652.30 ± 43.75 11,616.00 ± 31.19 9566.67 ± 22.50
pH 5.34 ± 0.11 5.74 ± 0.09 5.44 ± 0.06 5.34 ± 0.12 5.79 ± 0.11
Physicochemical property (HC.)
Viscosity (cPs) – – – – 9451.76 ± 16.97
pH – – – – 5.61 ± 0.40

Fig. 1. Extractive yield, total phenolics content and antioxidant activity of Moringa leaf extracts (* pyield < 0.05, † pantioxidant activity < 0.05, †† pTPC < 0.05).

as emollients, while dimethicone acted as an emollient and film-forming
agent. These W/Si emulsions were composed of water, sodium chloride
and Germaben® II functioning as diluent, stabilizer/adjuster and pre­
servative, respectively. The preparations were all white. Base D was
more slip than the others in regard with its least viscosity, while base B
was the thickest preparation. All of the base formulas (A-D) were proven
to be stable by means of the centrifugation assay. Of these, base D gained
the greatest preference as scaled by the formulator during the course of
product development. In view of the viscosity and texture of base D, it
was selected and incorporated with the antioxidative, phenolic-rich
Moringa leaf extract to give formula E. The preparation color shifted
from white to greenish yellow, and the preparation was slightly more
basic than the base D. The texture of the active emulsion was noted to be
improved in slip than the base D. The Moringa phytocosmetic product
was stable according to the centrifugation assay. The process was
thereafter scaled up and the preparation was evaluated with additional
accelerated stability test, i.e., heat-cool cycles. The developed formula­
tion was proven to be stable, as the viscosity and pH were insignificantly
shifted (p > 0.05).
4. Discussion
Moringa is regarded as a miracle tree with supportive biological
activities for health benefits (Oyeyinka and Oyeyinka, 2018). Moringa is
therefore widely cultivated, consumed and applied in cuisines and
health promotion recipes in accordance with their therapeutic activities,
of which glucosinolates are the known therapeutic actives of Moringa
(Stohs and Hartman, 2015) including phenolics that are of importance
for cosmeceutical products (Gunia-Krzyżak et al., 2018; Zillich et al.,
2015). The mature leaf has stronger antioxidant activity (with DPPH,
superoxide anion and nitric oxide; SOD and NO radicals, and lipid per­
oxidation assays) than the tender leaf. The therapeutic effects of this
medicinal plant are in accordance with its content of active phenolics
(Sreelatha and Padma, 2009). In addition, the antioxidative Moringa
leaf extract additionally inhibited microorganisms (Guillén-Román
et al., 2018) that are prohibited in topical products, i.e., Escherichia coli,
Pseudomonas aeruginosa and Staphylococcus aureus, including the body
malodorant and acne-associated microbe S. epidermidis (Kanlayavatta­
nakul and Lourith, 2011a;2011b). In the present study, mature Moringa
leaves were therefore examined objectively.
4.1. Preparation of antioxidative, phenolic-rich moringa leaf extract
Different solvents are crucial for isolating the active fraction of
Moringa leaves (Siddhuraju and Becker, 2003). The plant material was
first extracted with water for different durations in search of concise and

3
M. Kanlayavattanakul and N. Lourith
practical extractions feasible for industrial practice. Water was used as
the solvent because it was reported to yield the phenolic fraction of
Moringa leaf with an optimized 1:5 ratio of plant material and solvent
(Waterman et al., 2014).
The extractive yield was not clearly increased with longer extraction
durations (Fig. 1). Thereafter, ethanolic water was tested as the
extracting solvent instead of methanolic water (Siddhuraju and Becker,
2003) in view of the safety of methanol use, especially for health and
personal care products. The efficacy of alcoholic water as the extracting
solvent was assessed under ambient conditions and for different dura­
tions starting from 3 h that was exhibited as the most appropriated
duration of water extraction. It should be noted that alcoholic water
extractions with higher water contents and longer extraction durations
insignificantly increased the extractive yields (p > 0.05), as shown in
Fig. 1.
The quality of the extracts was therefore assessed comparatively by
means of the active principle content, TPC, and antioxidant activity. The
70% EtOH extract was significantly higher in TPC and antioxidant ac­
tivity (p < 0.05). Moringa leaf extracts with higher phenolics contents
are obviously more potent in biological activity studies (Guillén-Román
et al., 2018). This Moringa leaf extract was richer in TPC than Indian
cultivated plants (8.11 ± 0.06 and 4.58 ± 0.02 g GAE/100 g) (Sid­
dhuraju and Becker, 2003; Sreelatha and Padma, 2009). It should be
noted that this 70% EtOH extract of Moringa leaf was more potent in
DPPH radical scavenging (IC50 = 62.94 ± 7.05 and 47.2 − 149.8 µg/ml)
than previously reported (Guillén-Román et al., 2018, Vongsak et al.,
2013). Thus, this extract was highlighted as an appropriate anti­
oxidative, phenolic-rich Moringa leaf extract to be developed into a
phytocosmetic product.
4.2. Development of a stable W/Si emulsion containing moringa leaf
extract
Physicochemical properties of the cosmetic substances including
plant extract are crucial for the topical product development. Of which,
miscibility and pH are basic requirements for preparation of topical
agents. The extract was proved to be suitable for its use as a topical
product ingredient (Lourith and Kanlayavattanakul, 2017).
Topical products in emulsion dosages, i.e., water-in-oil (W/O) or oil-
in-water (O/W), rely on common carrier systems for dermatological and
cosmetic applications. Selection of the appropriate emulsifiers and
lipophilic and hydrophilic materials is key to success for the stability and
texture of the preparations. Nonetheless, these are difficult obstacles to
overcome in product development. Thanks to advancements in silicone
technology, W/Si emulsions have become more useful in terms of sta­
bility and textural preferences (Mittal and Bui, 2021; Tadros, 2018). The
W/Si emulsions were developed in the present study (Table 1). Variation
of dimethicone content significantly affected (p < 0.05) the textures of
the preparations and influenced viscosity (Calixto and Campos, 2017;
Gilbert et al., 2013). Base D with the greatest preference was incorpo­
rated with the antioxidative, phenolic-rich Moringa leaf extract to give
formula E. The proportion of the extract was tested at 0.5%, which
accounted 100-fold increase of its IC50, which was approximately 10
times the ascorbic acid activity described in the above section. The
developed formulation was proven to be stable, as the viscosity and pH
were insignificantly shifted (p > 0.05). The prepared antioxidative,
phenolic-rich Moringa leaf extract was therefore confirmed to be
compatible with available cosmetic ingredients, even though it was
tested at a high proportion.
5. Conclusions
Antioxidative, phenolic-rich Moringa leaf extract was successively
prepared. Maceration of the mature Moringa leaves in 70% EtOH for 24
h at room temperature afforded the greatest yield of phenolic extract
with prominent antioxidant activity. Information required for
4
Scientia Horticulturae 295 (2022) 110894
preparation of topical products was examined. The extract was shown to
be compatible with the available cosmetic ingredients, giving a stable
W/Si emulsion containing Moringa leaf extract. Preparation of topical
products containing the antioxidative, phenolic-rich extract of Moringa
leaf is presented herein and is available for safety and efficacy assess­
ments in human volunteers. Thus, a concise, feasible and scalable
extraction of antioxidative and phenolic-rich Moringa leaf fraction is
established. The innovative utilization of Moringa, a food crop, for
certain different industries such as cosmetics and personal care is
enabled. Successive benefits integrated in the agricultural and cosmetic
industries are evidenced. Sustainable/natural products derived from the
miracle tree Moringa are offered.
CRediT author statement
MK contributed to project planning and management, designed the
study, TPC analysis, development of the formulations and drafting of the
manuscript. NL contributed on extraction and antioxidant activity
assessment, data analysis and critical reviewing of the manuscript. All
the authors have read the final manuscript and approved the submission.
Funding
This research did not receive any specific grant from funding
agencies in the public, commercials, or not-for-profit sectors.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgments
The authors would like to acknowledge Assoc. Prof. Dr. Nijsiri
Ruangrunsri, Department of Pharmacognosy, Faculty of Pharmaceutical
Sciences, Chulalongkorn University, Bangkok, Thailand, for identifica­
tion of plant material. Mae Fah Luang University is acknowledged upon
facility supported during the manuscript preparation. The reviewers are
acknowledged on their valuable suggestions that make the article more
comprehensive.
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