Co esponding au ho : Thanda Aung
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion Liscense 4.0.
Ul asonic assis ed medicinal plan ex ac ion: The e iew
Thanda Aung * and S eemoy Kan i Das
Facul y o Pha macy, Lincoln Uni e si y College, Selango , Malaysia.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
Publica ion his o y: Recei ed on 21 June 2025; e ised on 26 July 2025; accep ed on 29 July 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.2.2811
Abs ac
The ex ac ion o bioac i e compounds om medicinal plan s plays a pi o al ole in he de elopmen o pha maceu ical,
nu aceu ical, and cosme ic p oduc s. Con en ional ex ac ion me hods, al hough widely used, o en su e om
limi a ions such as p olonged ex ac ion imes, high sol en consump ion, and deg ada ion o he molabile compounds.
Ul asonic-assis ed ex ac ion (UAE) has eme ged as a g een, e icien , and scalable al e na i e, le e aging acous ic
ca i a ion o enhance mass ans e and dis up plan cell walls. This e iew comp ehensi ely discusses he
undamen al p inciples o UAE, including he ole o ul asonic equency, powe in ensi y, sol en ype, and ex ac ion
du a ion. Compa a i e analysis wi h adi ional echniques demons a es UAE’s supe io i y in e ms o yield, ime
e iciency, and ene gy conse a ion. Recen ad ancemen s in eac o design, hyb id sys ems, and s a is ical
op imiza ion (e.g., esponse su ace me hodology) a e highligh ed. Limi a ions ela ed o equipmen scale-up, p ocess
ep oducibili y, and bioac i e s abili y a e c i ically add essed. Finally, u u e di ec ions include he use o g een
sol en s and AI-d i en modeling o indus ial-scale op imiza ion. This e iew p o ides esea che s and indus y
s akeholde s wi h an in-dep h unde s anding o UAE as a sus ainable, high-pe o mance echnology o he ex ac ion
o medicinal plan bioac i e.
Keywo ds: Ul asonic-Assis ed Ex ac ion (UAE); Medicinal Plan s; G een Ex ac ion Technologies; Ca i a ion;
Phy ochemicals; Bioac i e Compounds
1. In oduc ion
Fo many yea s, adi ional medicine has u ilized plan s o medicinal pu poses o cu e many illnesses. Cen au ium
e y h eae Pe s. - an i-in lamma o y, an ipy e ic, hypoglycemic, an ioxidan , an imic obial, hepa op o ec i e,
gas op o ec i e, e c. [1]; Glycy hiza glab a L. - an i-in lamma o y, an iulce , expec o an , an imic obial, and anxioly ic
ac i i ies [2]; Silene ulga is (Moench) Ga cke - good o b onchi is and as hma [4]; Aspala hus linea is L. - elie es
alle gies, de ma ological issues, as hma, in an ile colic, and o he gas oin es inal complain s, such as nausea and
hea bu n [5,6]; and Sambucus nig a L. - bene icial e ec s on blood p essu e, glycaemia educ ion, immune sys em
s imula ion, an i umo po en ial, inc ease in he ac i i y o an ioxidan enzymes in he blood plasma, including
glu a hione, and he educ ion o u ic acid le els [7].
None heless, a ema kable numbe o he bal plan s wi h he apeu ic bene i s ha e a highe le els o bioac i es, which
wo k in conce wi h he human body o p o ide special he apeu ic bene i s wi h ew nega i e side e ec s. Fo example,
ulsi, neem [8], ashwagandha [9], osema y [10], hymus on anesii [11], eucalyp us [12], u me ic oo [13], co iande
[14], and o he s a e common medicinal and he bal plan s ha con ain bioac i es.
Ul asonic-assis ed ex ac ion (UAE) has been shown o be a highly e ec i e echnique o boos ing yield while using
less sol en and educing hea damage [15]. Th ough sonic ca i a ion, ul asound, a non- he mal p ocess in ensi ica ion
echnique, imp o es ex ac ion by inc easing sol en pene a ion and solu e di usion [16]. In o de o maximize UAE,
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
1349
a iables including empe a u e, powe , and ime mus be changed. This can inc ease he yields o saponins. Finding
he ideal ci cums ances o maximal yield and pu i y is made easie wi h he use o esponse su ace me hodology (RSM)
[17]. Since sepa a ion and pu i ica ion concen a e hese bioac i e ing edien s and inc ease hei he apeu ic e icacy,
hey a e equally impo an [18].
F ee adicals a e uns able chemicals c ea ed by bo h in e nal p ocesses like me abolism and ex e nal ac o s like
pollu ion and UV ays. An ioxidan s a e molecules ha help p o ec he body's cells om hese ha m ul e ec s. The
oxida i e s ess ha ee adicals can cause can lead o aging, in lamma ion, cell damage, and a a ie y o illnesses, such
as cance , hea disease, and neu ological issues. o b eak down cell walls and imp o e he elease o bioac i e chemicals
by c ea ing sonic ca i a ion in he ex ac ion media [19]. In o de o gene a e high-quali y, a ge - ich chemical ex ac s
in sho e ex ac ion imes wi h li le o no use o o ganic sol en s, UAE is also essen ial o phenolic componen
ex ac ion om samples [20, 21].
The ocus o his e iew is on ye unexplo ed ul asound-assis ed echnologies ha ex ac bioac i es om widely
accessible medicinal plan s. In his e iew, we will add ess UAE's echnological ea u es, mechanis ic knowledge, c ucial
componen s, and bioac i es ho oughly and up- o-da e.
2. Ul asound assis ed ex ac ion
2.1. Ex ac ion mechanisms
The undamen al idea behind ul asonic ex ac ion echnology is o inc ease ex ac ion e iciency by using he
ca i a ion, mechanical, and he mal e ec s o ul asound o speed up he elease, di usion, and dissolu ion o use ul
compounds inside he cell. The ca i a ion phenomena is he esul o a medium being ipped apa by high-ene gy
ul asonic wa es in o nume ous iny ca i ies ha immedia ely shu , c ea ing an ins an aneous p essu e o up o millions
o a mosphe es. The agmen a ion ime is sho ened by he emendous p essu e c ea ed by he iny bubbles in
ca i a ion bu s ing, which causes he plan cell walls o sha e and he en i e o ganism o be comple ed in a single
ins an . Simul aneously, he ul asonic ib a ion imp o es in acellula subs ance elease, di usion, and dissolu ion,
which g ea ly inc eases ex ac ion e iciency [22–24] (Figu e 1). The chemical e ec o ca i a ion may be a ibu ed o
g ea e ee adical gene a ion, deg ading a ge molecules [25]. Mechanical o ces emo e bioac i e chemicals om
ege al issue in wo s ages. Mechanical e ec s allow sol en o pe mea e he cell wall and damage he acellula ma ix,
educing pa icle size [26]. Thus, he solu ion di uses as om plan issues o sol en wi h enhanced mass ans e
ac oss plan memb anes un il equilib ium is eached [27].
The p ima y pu pose o his ul asonic in ensi y is o ind in o ma ion abou he chemical and physical cha ac e is ics
o plan s ex ac . Low- equency ul asound, which has a subs an ial ib a ion e ec , ope a es a equencies be ween
20 and 100 kHz. La ge ib a ion o ce and enhanced mechanical e ec a e con eyed o he gene a o , which gene a es
high ene gy and aises he sys em's empe a u e while encou aging mass ans e . Addi ionally, he use o ul asound
ea men has high epea abili y, low dependence on eagen s, and simple subsequen p ocessing o he p oduc [28].
Ul asound has many bene i s in ood p ocessing, including mo e ho ough mixing o a ious subs ances, lowe ing
p ocessing empe a u e, p omo ing mass ans e , sho ening p ocessing ime, and inc easing ex ac ion e iciency.
Figu e 1 Ul asonic-assis ed ex ac ion echnology [24]
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
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2.2. Ul asound de ices
Conside ing he e ec s so a , ul a sound may be a po en ial echnique o in oduce equipmen in o ex ac ion
p ocedu es. Samples can be a ec ed di ec ly o indi ec ly by ul asound. Di ec sound wa eleng h impac s occu
di ec ly on samples, while indi ec e ec s occu h ough con aine s be o e eaching samples. Some in es iga ions ound
ha ul asound p obes (ho ns) inc eased mass ans e . The absence o ul asound p opaga ion impedimen s may
explain hese imp o emen s. Ul asound p ocesses, usually done in ul a sound ba hs o powe ul ul asound eac o s,
indi ec ly a ec samples. The sample con aine had lowe ul asound in ensi y han expec ed because ul asound wa es
had o pass h ough he liquid in he ul asonic de ice and hen h ough he con aine wall [29]. Some cu en
applica ions equi e an ul asonic p obe ins ead o an ul asound ba h.
The comp ession and a e ac ion cycles (high and low-p essu e egions) ha comp ise ul asound wa es can a el
h ough solid, liquid, o gaseous media, dislodging and disloca ing molecules om hei ini ial posi ions and p oducing
eno mous amoun s o ene gy [30]. Fu he mo e, ca i a ion bubbles appea when high-in ensi y nega i e p essu e is
applied because he a e ac ion cycle exceeds he liquid molecules' a ac ion o ces [31]. While epheme al ca i a ion
o ine ial ca i a ion bubbles pe sis o a e y b ie pe iod, equen ly less han a single cycle, and hen apidly
disin eg a e, s able ca i ies, which a e long-las ing gas bubbles, a e p oduced by se e al comp ession and a e ac ion
cycles o sonica ion [32].
2.2.1. The ul asonic ba h
Ul asonic ba hs a e e o less o use and cos -e ec i e, bu he sol en and expe imen al glasswa e g ea ly educe he
ul asonic s eng h, making hem unsui able o chemical eac ions [33]. Indi ec ex ac ion gi es good esul s in he
ul asonic ba h, bu only a modes numbe o bioac i e subs ances may be ex ac ed. Di ec ex ac ion yields mo e.
O he esea che s ound ha pa icle size, acid and/o oxidan used, leaching olume, sonica ion ime, wa e
empe a u e inside he ba h, ul asonic equency, sample posi ion ( e ical o ho izon al) and de e gen use in he
wa e g ea ly a ec he sonica ion ba h [34, 35]. The abo e ac o s imp o e ul asound ansmission. Ul asound ba h
sys ems also ha e an ul asonic sounde ha gene a es an exci a ion signal ha ma ches he uning assembly's modal
equencies.
The ex ac ion cell design, which educes ul asonic powe , is he ha des pa o any ul asound-assis ed sys em. In
ba h- ype sonica ion, he hick walls o he ex ac ion cell bea he p essu e, a enua ing ul asonic wa es om he
ansduce s and limi ing in luence on he ex ac ion medium [36]. Piezoelec ic ansduce s u n elec ici y in o
mechanical ib a ions. In non-con ac applica ions, uning de ices ans e mechanically gene a ed ul asonic wa es o
ai bo ne ma e ial. Ul asound ansmission depends on he p opaga ion medium's acous ic impedance. The di e ence
be ween he ul asonic ansduce ma e ial's impedance and ai 's g ea ly a ec s non-con ac ul asonic wa e use.
2.2.2. The ul asonic p obe
The p obe sys em is commonly used o sonica e iny samples. Because ul asonic s eng h is con eyed exclusi ely a
he p obe ip, p obe- ype ul asound is s onge han ul asound ba h. P obe sonica ion aises sample empe a u es
sha ply. Thus, ul asonog aphy equipmen design ma e s. In ea ly ul asonic chemis y, an open beake was imme sed
di ec ly in an ul asonic ba h and he in luence o ul asonic p obes on ep oducibili y was examined [37]. Wi h
imp o ed ul asound gene a o s, he piezoelec ic ansduce could di ec ly en e he ul asonic sol en . This de ice
accele a es mass ans e bu inc eases sample des uc ion due o ul asonic p obe deg ada ion [38, 39]. Gi en he
uni o mi y o he equipmen 's p essu e ield, he eac o size and elemen ela i e o he senso mus be calcula ed o
maximize ene gy ans e o luid. This op imizes ex ac ion e iciency. Mapping can enhance ca i a ion o es eac o
dimensions and ansduce posi ions.
Ul asonic p obes p o ide mo e ul asound powe han ul asound ba hs because hey di ec ly subme ge in he solu ion
o sonica ion and imp o e ma e ial con ac a ea o educe mass ans e esis ance. No e ha ce ain se ings may al e
he ul asonic p obe: ampli ude, empe a u e, eac ion essel shape, diame e , uni , p obe ype, and ma e ial chemical
cha ac e is ics. The ul asound ca i a ion e ec is also a ec ed by he esponse essel o m, which mus be conical and
ha e a small diame e o aise he le el and allow he p obe o be inse ed deepe in o he sample. Due o oaming and
co osion in some samples, sono ode ma e ial and diame e a e c ucial. Hea - and co osion- esis an i anium alloy
p obes a e common [40]. Howe e , he ul asonic p obe can di ec ly con ac he sample in he p essu ed essel [41].
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
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3. Ul asonic assis ed medicinal plan ex ac ion
Ul asonic-assis ed ex ac ion (UAE) has eme ged as a powe ul g een echnology o enhancing he yield and
bioac i i y o phy ochemicals om medicinal plan s. Va ious s udies ha e op imized UAE pa ame e s o imp o e
ex ac ion e iciency, compound s abili y, and biological ac i i y.
Wang e al. [42] op imized he ul asonic-assis ed ex ac ion o u in om Ilex asp ella using deep eu ec ic sol en s
(DES), applying esponse su ace me hodology (RSM). Fac o s such as sol en ype, mola a io, wa e con en , liquid–
solid a io, ul asonic empe a u e, powe , and ime we e s udied. A Box–Behnken design iden i ied op imal condi ions:
lac ic acid o choline chlo ide mola a io o 1:1, 40 °C ex ac ion empe a u e, 31 min du a ion, 28% wa e con en , and
a 20:1 mL/g liquid–solid a io. Unde hese pa ame e s, u in yield was 86.553 ± 1.35 μg/g wi h <0.6% e o om he
p edic ed alue. Guan e al. [43] ex ac ed a neu al polysaccha ide (PLP-2-1) om Pe illa lea es unde op imal UAE
condi ions, ollowed by sul a ion o ob ain S-PLP-2-1. S uc u al analyses (HPLC, me hyla ion, NMR) con i med high
simila i y pos -modi ica ion. Ze a po en ial, FTIR, he mog a ime ic, and heological analyses suppo ed
cha ac e iza ion. Bo h o ms showed an ioxidan and hypoglycemic ac i i ies. In i o es s on oleic acid-induced HepG2
cells e ealed imp o ed lipid p o iles, wi h enhanced biological e ec s obse ed a e sul a ion, sugges ing he apeu ic
po en ial agains glycolipid me abolic diso de s.
Younis e al. [44] in es iga ed Sopho a japonica ex ac s ob ained ia UAE and o mula ed hem in o nanoemulsions.
Compa ed o mace a ion (51.18 mg GAE/mL), UAE yielded highe phenolic con en (65.57 mg GAE/mL), demons a ing
supe io an ioxidan capaci y. Legesse e al. [45] op imized UAE o Ve bascum sinai icum, a ge ing phenolic
ex ac ion. Va iables included ex ac ion ime, empe a u e, and sample–sol en a io. UAE ou pe o med mace a ion
in yield (21.6%), o al phenolics (179.8 mg GAE/g), la onoids (64.49 mg CE/g), and an ioxidan ac i i ies (DPPH:
61.85 μg/mL, ABTS: 38.89 μg/mL). UHPLC-ESI-QTOF-MS/MS iden i ied 17 phenolics, and SEM con i med
mic os uc u al changes. Op imal condi ions we e 41.43 °C, 36.32 g/mL sample–sol en a io, and 33.22 min ex ac ion
ime, suppo ing UAE’s applicabili y in pha maceu ical de elopmen . Yin and Ba ba an [46] es ablished op imal UAE
pa ame e s o Po ulaca aizoon, achie ing a la onoid yield o 10.77 mg/g unde 1 g/55 mL solid–sol en a io, 60%
e hanol, 45 °C, 25 min, and 150 W ul asound powe . The high yield was a ibu ed o ex ended ex ac ion o e h ee
days.
Zhang e al. [47] used enzyme-assis ed UAE o ex ac an hocyanins om Xinjiang mulbe ies and de eloped p edic i e
models using RSM and deep neu al ne wo ks (DNN). Va iables included pec inase dosage, hyd olysis ime, ul asonic
condi ions, sol en concen a ion, and solid–liquid a io. Bo h models we e e ec i e, bu DNN showed supe io
accu acy (R² = 0.9900, e o = 0.85%) compa ed o RSM (R² = 0.9404, e o = 4.50%), highligh ing DNN’s ad an age o
p ecise p edic ion o op imal ex ac ion ou comes.
Table 1 compiles key s udies on ul asonic-assis ed ex ac ion (UAE) o medicinal plan s, de ailing me hodologies,
ex ac ion pa ame e s, and yields. I highligh s how ac o s like ul asonic powe , empe a u e, and ime in luence he
e iciency and e ec i eness o bioac i e compound eco e y.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
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Table 1 Ul asonic Assis ed Medicinal Plan Ex ac ion
Re e ences
Medicinal
Plan
Me hodology
Ul asonic
powe
Ul asound
empe a u e
Ex ac ion
ime
Ex ac ion
yield
Wang e al.
[42]
Ilex asp ella
Box-Behnken
Response su ace
me hod
100 W
40 °C
31 min
-
Guan e al.
[43]
Pe illa lea es
Response su ace
me hod
100 W
60∘C
6 h
1.14%
Younis e al.
[44]
S. japonica
lowe s
E hanolic mace a ion
me hod
600 W
25 ℃
15 min
-
Legesse e al.
[45]
Ve bascum
sinai icum
(Qe e ina o
yeahya Jo o)
RSM and Mace a ion
ex ac ion
750 W
41.43 ∘C
33.22 min
21.6%
Li e al. [48]
Pla ycodon
g andi lo um
oo s
Response su ace
me hod (RSM)
153.79 W
49.59 ℃
51.04 min
4.83 %
Chen e al.
[49]
Tu me ic
(Cu cuma longa
L.)
Ionic liquid
ul asound-assis ed
hyd odis illa ion
-
50℃
37 min
6.88 %
Souadia e
al. [50]
Thymus
alge iensis
Box-Behnken design
(BBD) o esponse
su ace me hodology
(RSM)
-
60∘C
36.74 min
-
Zhou e al.
[51]
Ble illa s ia a
In o ma ion-
dependen
acquisi ion me hod
350 W
5∘C
20 min
-
Saeed Abadi
e al., [52]
Common
ho se ail
(Equise um
a ense L.)
RSM
-
27.88 °C
6.91 min
-
Lee e al.
[53]
Phedimus
aizoon
UAE
1500 W
45∘C
120 min
16.56%
Wang e al. [24] epo ed ha Muxu con ains o e 80 la onoid- ela ed compounds, including la onoids and hei
glycosides, la onols, iso la ones, chalcones, dihyd o la ones, and osewood-de i ed compounds. F om chlo o o m and
n-bu anol ex ac s o Muxu, se en compounds we e isola ed and iden i ied: apigenin 7-O-β-D-glucoside, u idine, β-D-
glucoside, lu eolin-7-O-me hyl glucopy anose, apigenin, Muxu ex ac , and 7,4’-dihyd oxy la one. Younis e al. [43]
e alua ed he an ioxidan ac i i y o Sopho a japonica ex ac s using he DPPH assay. The ul asound-assis ed ex ac
demons a ed a 67% sca enging e iciency, ou pe o ming he e hanolic ex ac 's 59%. Nanoemulsions o mula ed om
he ul asound ex ac s showed an a e age pa icle size o 252.92 nm and a ze a po en ial o −36.68 mV, indica ing high
s abili y. O e a 5-day oxida i e s abili y es , wa e -based nanoemulsions main ained s uc u al in eg i y, while
e hanolic o mula ions exhibi ed phase sepa a ion and inc eased oxida ion.
Zhang e al. [47] de e mined he op imal condi ions o an hocyanin ex ac ion om mulbe ies using a deep neu al
ne wo k (DNN) model: a solid–liquid a io o 50 mL/g, e hanol concen a ion o 63%, ul asonic empe a u e o 40 °C,
and pec inase dosage o 0.5%. Unde hese condi ions, o al an hocyanin con en eached 3.16 mg/g. An ioxidan
e alua ions showed DPPH, ABTS, and hyd oxyl adical sca enging a es o 80%, 98%, and 54%, espec i ely,
highligh ing he po en an ioxidan capaci y o mulbe y an hocyanins and demons a ing a sus ainable, op imized
ex ac ion me hod. Qi e al. [54] used la onoid ex ac ion om Po ulaca aizoon and assessed an ioxidan capaci y
h ough ABTS and DPPH adical sca enging. A 0.3 mg/mL, P. aizoon ex ac and i amin C showed ABTS sca enging
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
1353
a es o 96.2% and 99.0%, espec i ely. A 2.0 mg/mL, DPPH sca enging eached 85.9% o he ex ac and 98.8% o
i amin C, demons a ing s ong ee adical neu alizing abili y o he plan ex ac , compa able o asco bic acid.
Hi anp adi h e al. [55] op imized UAE pa ame e s o Cen ella asia ica o maximize o al phenolic con en (TPC) and
o al la onoid con en (TFC) using esponse su ace me hodology (RSM). E hanol concen a ion and sol en olume
signi ican ly in luenced bo h TPC and TFC (p < 0.0001), while ul asonic powe had minimal impac (p < 0.05).
Ex ac ion ime did no signi ican ly a ec TPC (p > 0.05) bu in luenced TFC due o la onoid deg ada ion (p < 0.05).
Va iable in e ac ions we e s a is ically insigni ican (p > 0.05). The i ed quad a ic models achie ed p edic ed R² alues
o 0.8263 o TPC and 0.9006 o TFC. The op imal condi ions—75% e hanol, 87.5 W ul asonic powe , 30 min
ex ac ion ime, and 20 mL sol en olume—yielded TPC and TFC o 52.29 ± 1.65 mg/g and 43.71 ± 1.92 mg/g,
espec i ely. Addi ionally, Asia ic side and madecassoside yields eached 37.56 ± 4.25 mg/g and 16.91 ± 1.28 mg/g.
These indings unde sco e he e iciency, scalabili y, and sus ainabili y o UAE o ex ac ing bioac i e om C. asia ica,
suppo ing i s use in unc ional ood de elopmen .
Table 2 summa izes he applica ion o UAE ac oss a ious medicinal plan s, highligh ing op imized ex ac ion
condi ions such as sol en a ios and analy ical me hods. The able also demons a es UAE’s e ec i eness in enhancing
an ioxidan ac i i y, con i ming i s po en ial o high-yield bioac i e eco e y.
Table 2 Applica ions o Ul asonic-Assis ed Ex ac ion (UAE) o Bioac i e Compounds om Medicinal Plan s:
Ex ac ion Condi ions and An ioxidan Ac i i ies
Re e ence
Medicinal Plan
E alua ion o UAE Ex ac s
Ex ac ion Ra io
An ioxidan
Ac i i y
Wang e al.
[42]
Ilex asp ella
Ru in yield: 86.553 ± 1.35 µg/g
Liquid–solid a io
o 20:1 mL/g
–
Guan e al.
[43]
Pe illa lea es
HPLC, me hyla ion analysis, NMR
Solid–liquid a io
o 1:30 (w/ )
74.19%
Younis e al.
[44]
Sopho a japonica
lowe s
DPPH assay; TPC: 65.57 mg GAE/mL
–
67%
Legesse e al.
[45]
Ve bascum
sinai icum
TPC: 179.8 mg GAE/g, TFC: 64.49 mg
CE/g, DPPH: 61.85 µg/mL, ABTS: 38.89
µg/mL
Sample–sol en
a io: 36.32 g/mL
38–61%
Li e al. [48]
Pla ycodon
g andi lo um oo s
An ioxidan ac i i y assay
Equal olume
a io (1:1)
70%
Chen e al.
[49]
Cu cuma longa L.
(Tu me ic)
Ionic liquid UAE-hyd odis illa ion (IL-
U-HD)
Liquid–ma e ial
a io o 12:1
22%
Lee e al. [53]
Phedimus aizoon
GC-MS and HPLC ch oma og aphic
analyses
–
24–28%
4. E alua ion o UAE Ex ac s
4.1. Response Su ace Me hodology and Op imiza ion o UAE
The ideal condi ions o ex ac ing he o al phenolic con en (TPC) om V. Sinai icus we e ound in 20 expe imen al
uns by es ing independen a iables including sonica ion ime, sol en - o-solu e a io, and ex ac ion empe a u e in
he UAE. A second-o de polynomial equa ion was ound o he esponse a iables as ollows
𝑌𝑛= 𝛽0+ 𝛽1𝑥1+ 𝛽2𝑥2+ 𝛽3𝑥3+ 𝛽11𝑥1
2+ 𝛽22𝑥2
2+ 𝛽33𝑥3
2+ 𝛽12𝑥2
2+ 𝛽13𝑥3
2+ 𝛽23𝑥3
2 …. (1)
whe e 𝑌
𝑛 (TPC) is ep esen ed by he a iables 𝛽0, 𝛽𝑖, 𝛽𝑖𝑖 and 𝛽𝑖𝑗. These a e he in e cep , linea eg ession coe icien
o he 𝑖- he componen , quad ic, and in e ac ion e ec e m, and he independen a iables a e coded as 𝑋𝑖 and 𝑋𝑗 .The
numbe o a iables being examined is deno ed by 𝑘 [45].
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4.2. To al Phenolic Compounds
The o al phenolic con en was calcula ed using he Folin-Coal e p ocedu e wi h minimal modi ica ions [56]. In a 15
mL cen i uge ube, 5 mL o ex ac and 5 mL o 80% me hanol we e mixed oge he . A e ha , he ubes we e
cen i uged o 20 minu es a 4000 pm a 4–16KS, Ge many. Fo analysis, 3000 L o deionized wa e , 100 L o Folin
Coal e eagen , and 100 L o a s anda d solu ion o sui ably dilu ed sample (10−100 g/mL) we e mixed oge he and
o exed. A e 10 minu es o oom empe a u e incuba ion, 100 L o a 20% sodium ca bona e solu ion was added igh
away, s i ed, and allowed o si a oom empe a u e o wo hou s in he da k. The abso bance o he combina ion a
765 nm was hen measu ed using a mic opla e eade (Bio ech Syne gy 2 Mic opla e eade , USA). Using millig ams o
gallic acid as he e e ence, he o al phenolic con en s o he samples we e ep esen ed as millig ams o gallic acid pe
100 millili e s.
4.3. To al Fla onoids Con en (TFC)
TFC was assessed using Zhi’s hen e al.'s me hodology [57]. Fi s , 2.5 mL o dis illed wa e , 0.15 mL o 5% sodium ni i e,
and 0.5 mL o he ex ac we e mixed oge he . A e 6 minu es o s anding ime, 0.3 mL o 10% m/V aluminum chlo ide
was added o he mix u e and p ope ly mixed. This was ollowed by he addi ion o 0.55 mL o dis illed wa e and 1 mL
o 1.0 M sodium hyd oxide. A e ha , he mix u e was o exed and allowed o s and o i een minu es. U ilizing a UV-
Vis spec opho ome e (Op i Zen 2120UV; Me asys Co., L d., Daejeon, Republic o Ko ea), he concen a ion was las ly
measu ed a 510 nm. Fo quan i ica ion, a calib a ion cu e o ca echin was c ea ed, and he esul s we e gi en as (mg
CE/g DW) d y ex ac o he sample.
4.4. To al Polyphenol Con en
The Folin-Coal e app oach was used o calcula e TPC calo ime ically [58]. 2.5 mL o 10% Folin-Coal e eagen (FCR)
was combined wi h 0.2 mL o he ex ac o c ea e he solu ion. Two millili e s o a 7.5% sodium ca bona e solu ion
con aining 75 g/mL was hen added. A e 10 minu es o hea ing o be ween 50 and °C, he sample was le o cool. A
Spec a i3x pla e eade (Molecula De ices, LLC., Seoul, Republic o Ko ea) was used o measu e he abso bance a 750
nm. Gallic acid s anda d was used o c ea e a calib a ion cu e, and he esul s we e epo ed in mg GAE/g dew.
4.5. DPPH Sca enging Assay
The abili y o sca enge ee adicals while in e ac ing wi h a s able DPPH ee adical was used by Ciconine, Iadanza’s,
and Vikas [59] o e alua e an ioxidan ac i i y.
Using Equa ion (2), an ioxidan ac i i y was compu ed as a pe cen age o adical sca enging.
% adical sca enging pe cen age =A0 −A1
A0 ×100 …. (2)
The abso bance o he con ol eac ion, which con ains all eagen s excep he es subs ance [𝑡=0 min], is deno ed by
A0. (𝑡=30 min) Al = abso p ion o es ex ac solu ion. A e 30 minu es o incuba ion, he ligh abso p ion a oom
empe a u e was measu ed a 517 nm. Wi h compa able AOA epo ed as 𝜇M T olox pe g DW, he DPPH adical
sca enging ac i i y was shown as a unc ion o T olox concen a ion.
4.6. ABTS adical ca ion decolo iza ion assay
The ex ac s' abili y o sca enge adicals agains adical ca ions (ABTS, -++.) was calcula ed using a p e iously published
me hod wi h mino adjus men s [60]. In e ms o 𝜇M TE/g DW, he esul s we e p esen ed.
4.7. Fe ic educing an ioxidan powe assay (FRAP)
The FRAP assay was pe o med in acco dance wi h Benzie and S ain's p o ocol [61]. E e y day, he FRAP eagen was
made om sc a ch and hea ed o 37 °C be o e use. Following ou minu es o incuba ion a 37 °C, he abso bance o he
eac ion mix u e was measu ed a 593 nm. 𝜇MTE/g DW was used o exp ess he esul s.
4.8. The mog a ime ic Analysis
The he mog a ime ic analysis (TGA) o he 70% e hanol ex ac o S. japonica UAE was ca ied ou using an SDT Q600
(V20.9 Build 20) and he nano emulsion (ENE). A sample o abou 20 millig ams was quan i ied and ca e ully packed in
aluminum con aine s. The samples we e lushed wi h ni ogen a a a e o 30 mL/min while being aised om 30-40°C
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o 600-°C a a a e o 10-°C/min. A me al con aine ha was emp y se ed as he e e ence [62]. The impac o e hanol
on s abili y was assessed by compa ing he sample wi h he UAE 100% wa e ex ac nano emulsion (WNE).
4.9. Colo ime ic Analysis
A colo ime e was used o assess he ex ac s' colo [63]. The equa ion ha was used o de e mine he o e all
ch oma ici y di e ence (𝐸) used he alues o 𝐿∗ (ligh ness), 𝑎∗ ( edg een), and 𝑏∗ (yellow-blue):
Δ𝐸 = (Δ𝐿∗2 + Δ𝑎∗2 + Δ𝑏∗2)1
2 … … … .. (3)
The alues o Δ𝐿, Δ𝑎 , and Δ𝑏 we e calcula ed by sub ac ing he s anda d colo pa ame e s om he
co esponding, 𝐿∗, 𝑎∗, and 𝑏∗ alues. The squa e oo o he sum o he squa ed di e ences in, 𝐿∗, 𝑎∗, and 𝑏∗. was hen
used o compu e he o al ch oma ici y di e ence (Δ𝐸). Whi eboa d alues we e calib a ed as 𝐿∗=89.03; 𝑎∗=
0.35; 𝑏∗= 3.42.
4.10. X-Ray Di ac ion
A Ge man B uke D8 Ad ance x- ay di ac ome e (XRD) was used o examine he c ys al s uc u es o se e al d y
samples. P io o es ing, he samples we e la ened and posi ioned in he middle. The scanning ange o 2𝜃 was
es ablished a a a e o 5∘ pe minu e and was se o 5∘−40∘. To de e mine he c ys al s uc u e, he di ac ion pa e ns
we e ga he ed and examined [64].
4.11. A enua ed To al Re lec ance-Fou ie T ans o m In a ed
An FTIR spec ome e (Nicole iS50, The mon Elec on, USA) i ed wi h a diamond c ys al a enua ed o al e lec ion
(ATR) accesso y was used o acqui e he Fou ie ans o m in a ed (FTIR) spec a o d y ma e ials. Wi h a esolu ion
o 4, cm-1, and a p ede e mined numbe o scans o 16, he spec a we e ob ained be ween 4000 and 500 cm-1 [65].
4.12. Con ocal Lase Scanning Mic oscopy
Using a con ocal lase scanning mic oscope (CLSM) (Leica TCS SP5, Ge many), he emulsions' mo phology was
desc ibed. The mic omo phology o he emulsion was examined using CLSM. A a empe a u e o 25 ℃, 10 L o he
emulsion was speci ically placed on a mic oscope slide, co e ed wi h a co e slip, and examined wi h a 100× oil
imme sion objec i e lens. E e y pic u e was aken wi h a scanning equency o 200 Hz and a scanning densi y o 1024
× 1024 [66].
5. Conclusion
Ul asonic-assis ed ex ac ion (UAE) ep esen s a signi ican ad ancemen in he ield o g een ex ac ion echnologies,
o e ing an e icien , cos -e ec i e, and en i onmen ally iendly al e na i e o con en ional me hods. By ha nessing
acous ic ca i a ion, UAE enhances mass ans e and dis up s plan cell ma ices, esul ing in imp o ed ex ac ion yields
o phenolics, la onoids, and o he bioac i e wi h educed sol en usage and sho e ex ac ion imes. The applica ion
o s a is ical op imiza ion echniques such as esponse su ace me hodology (RSM) has u he e ined UAE pa ame e s,
enabling a ge ed eco e y o speci ic compounds. Analy ical e alua ions, including DPPH, ABTS, FRAP assays, and
s uc u al analyses (HPLC, NMR, FTIR, XRD), a i m he supe io an ioxidan capaci y and s abili y o UAE-de i ed
ex ac s. Despi e i s many ad an ages, challenges such as equipmen scale-up, s anda diza ion, and compound
deg ada ion unde high-in ensi y sonica ion emain. Fu u e esea ch should p io i ize he in eg a ion o g een sol en s,
eal- ime moni o ing echnologies, and a i icial in elligence-based modeling o u he op imize UAE o indus ial
applica ions. O e all, UAE holds emendous p omise o sus ainable and scalable ex ac ion o bioac i e compounds
om medicinal plan s, suppo ing i s b oad u ili y in unc ional ood, pha maceu ical, and cosme ic indus ies.
Compliance wi h e hical s anda ds
Disclosu e o con lic o in e es
No con lic o in e es o be disclosed.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1348-1359
1356
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