Co esponding au ho : Ligia ION
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.
Resea ch on he ca bohyd a e and p o ein con en o basil geno ypes om he Buzău
Region, Romania
Mădălin Vasile CHIUOARU, Lenuța CHIRA, Daniel NAGY, Gab iela BUTNARIU and Ligia ION *
Uni e si y o Ag onomic Sciences and Ve e ina y Medicine o Bucha es , 59 Ma as i Bl d, Dis ic 1, Bucha es , Romania.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1102-1110
Publica ion his o y: Recei ed on 27 June 2025; e ised on 11 Augus 2025; accep ed on 13 Augus 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.2.2936
Abs ac
The a oma ic cha ac e o each ype o basil is de e mined by he geno ype and depends on he majo chemical
compounds o med mainly by he chemical compounds mono e penes and essen ial oils. phenylp opanoids. The
essen ial oil has an imic obial, an i ungal and insec epellen , an icon ulsan , hypno ic and an ioxidan ac i i ies.
Resea ch has shown ha he Basil A oma ic o Buzău geno ype showed a signi ican ly posi i e di e ence in e ms o
ca bohyd a e con en , acco ding o ANOVA analysis. The o he geno ypes (L1, L7, L8), al hough hey showed posi i e
di e ences, hese we e no la ge enough o be s a is ically signi ican . This analysis suppo s he choice o he Basil
A oma ic o Buzău geno ype o c ops o ien ed owa ds a high ca bohyd a e con en . The p o ein con en alues we e
in luenced by he geno ype and he en i onmen al condi ions in he yea s in which he esea ch was ca ied ou . The
A oma ic basil o Buzău geno ype eco ded he highes p o ein con en , and i s alues signi ican ly exceeded he DL,
which con i ms ha he di e ences om o he geno ypes a e s a is ically signi ican .
Keywo ds: Ocimum basilicum; Geno ypes; Ca bohyd a e; P o ein; S a is ical Analysis
1. In oduc ion
The plan Ocimum basilicum o igina es om ancien Pe sia, Pakis an and India, as well as o he a eas in Asia. Basil
g ows op imally in a empe a e clima e, ha ing a low sensi i i y o low empe a u es and p e e ing he Medi e anean
egions. I g ows spon aneously in Asia, A ica and he wa m a eas o No h Ame ica. In opical clima ic condi ions, he
plan de elops o e a pe ennial pe iod o se e al yea s, while in empe a e a eas, i beha es as an annual species. [11],
[8]. Basil has an isep ic p ope ies o he in es ines, kidneys, eb i uge, an i ungal, galac ogogue, diu e ic, an ieme ic
and an i-in lamma o y o bo h he kidneys and in es ines, e c. I des oys bac e ia in he body, especially in es inal ones
and hose in he kidneys and lungs. [10], [12], [6] I ac s agains pa asi ic ungi ha a ec he body, educes e e ,
elimina es abdominal pain and nausea, while s imula ing milk sec e ion in women, e c. Among he condi ions ea ed
in animals a e kidney diseases, cys i is, including hemo hagic cys i is, and gas oin es inal p oblems. Basil is
ecognized as an impo an s o ehouse o bene icial subs ances, also ha ing applicabili y in mode n medicine.
Basil con ains impo an i amins, namely: i amin A (p o ec i e ole o he eyes and mucous memb anes, helps
main ain heal hy skin), i amin K (plays an impo an ole in blood clo ing and helps s eng hen bones). Vi amins B1,
B2, B3, C and E a e also p esen in basil. Species o he genus Ocimum do no show ha m ul e ec s, as long as hey a e
used co ec ly in he apeu ic ea men s. The e o e, in case o o e dose, poisoning may occu , which o en p esen s
wi h symp oms o con ac de ma i is. Due o i s u e o onic p ope ies, he oil is no ecommended o use du ing
p egnancy, bu i can be applied du ing massage (1-2 d ops added o a massage oil). [4], [1], [3], [7].
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Basil has been used in adi ional medicine in Sou h Asia o o e h ee millennia. O e ime, i has acqui ed he s a us
o a mul i unc ional medicinal plan , ecognized and alida ed in con empo a y s udies by esea che s and doc o s. In
he las decade, he mul iple he apeu ic e ec s o basil ha e been con i med: adap ogenic, an imic obial, an i-
in lamma o y, ca dio- and hepa op o ec i e, immunomodula o y, me abolic an icance , an ioxidan , adiop o ec i e,
an idiabe ic, an ispas ic. [2], [5], [9].
2. Ma e ial and me hods
The plan ma e ial used in he s udy consis s o basil geno ypes om sou he n Romania, in su icien numbe s o
conduc a meaning ul s udy o he g ow h cha ac e is ics o basil geno ypes (Table 1.) included in he s udy:
Table 1 Plan ma e ial basil plan biome ics o he 16 geno ypes s udied (su as/sou ce o iginal)
N ca .
Geno ypes s udied
A e age heigh o he plan (cm)
1.
A oma ic Basil om Buzau
75.50
2
Pu ple basil Se a im
77.10
3
Swee basil L1
62.20
4
Lemon basil L2
73.30
5
G eek basil
30.21
6
Basil wi h salad lea es L4
84.40
7
G and Ve Basil L5
60.30
8
Pe sian basil
52.48
9
Spicy Globe Basil L7
37.51
10
A ican basil L8
118.00
11
Holy basil a . K ishna L9
78.10
12
Macedonian basil
60.85
13
Dwa basil Sma ald
29.40
14
Da k Opal ed basil
56.56
15
Siam Queen Basil
51.48
16
Basil cinnamon
110.50
2.1. Ope a ional me hod
The de e mina ion o ca bohyd a e con en in he basil a ie ies s udied was ca ied ou by he colo ime ic me hod
(phenol–sul u ic acid me hod).
P inciple o he phenol–sul u ic acid me hod (colo ime ic me hod o o al ca bohyd a es). Ca bohyd a es in he plan
ex ac a e subjec ed o dehyd a ion in a concen a ed acid medium (sul u ic acid). Unde he ac ion o sul u ic acid,
suga s a e con e ed in o u u al o i s de i a i es. These compounds eac wi h phenol, o ming a yellow–o ange
colo ed complex. The colo in ensi y is di ec ly p opo ional o he ca bohyd a e concen a ion and is de e mined
spec opho ome ically a λ = 490 nm. (Figu e 1.1.)
2.2. The wo king s eps we e
Sample p epa a ion – he plan ma e ial (basil lea es) was d ied, g ound, hen he ca bohyd a e was ex ac ed in
dis illed wa e o 80% e hanol.
• Addi ion o phenol (5% solu ion).
• Addi ion o concen a ed sul u ic acid o hyd olyze and dehyd a e ca bohyd a es.
• Fo ma ion o he colo ed complex ( he eac ion was allowed o p oceed o 10–20 min).
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1102-1110
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• Reading he abso bance on he spec opho ome e a 490 nm.
• Calcula ion o he ca bohyd a e con en based on a s anda d cu e ob ained wi h a s anda d glucose solu ion.
Wo king p inciple o he Kjeldahl me hod ( o al ni ogen → o al p o ein de e mina ion)
Fo he de e mina ion, he d ied samples we e diges ed in concen a ed sul u ic acid wi h a ca alys ; he o ganic
ni ogen was con e ed o ammonium; he ammonium was hen dis illed as NH₃ a e alkaliza ion and hen i a ed.
The ni ogen con en is con e ed o p o ein by a con e sion ac o (usually 6.25 o o al p o ein, bu o plan s 5.7 can
also be used — he me hod is speci ied in he pape ).
2.3. Ma e ials and Reagen s
• Concen a ed H₂SO₄ (98%)
• Ca alys (K₂SO₄ + coppe o me cu y, o selenium sul a e)
• 40% sodium hyd oxide (NaOH) o alkaliza ion
• 4% bo a e solu ion (o collec ion solu ion o dis illa ion)
• N HCl (o s anda dized H₂SO₄) o i a ion
• Phenolph halein indica o o p e e ed i a ion solu ion
2.4. Sample p epa a ion
• Ha es lea es om all 16 basil geno ypes s udied (same phenological s age), d y a 60–70 °C o cons an
weigh .
• G ind d y in a pes le and homogenize.
• Reco d he mass o he d y sample (0.5–1.0 g, depending on he N con en and sensi i i y o he me hod).
2.5. P ocedu e ( ypical p o ocol)
In he diges ion ube/e lenmeye lask, 0.5–1.0 g o d y sample + 10–15 mL o concen a ed H₂SO₄ + 1–2 g o K₂SO₄ +
0.5 g o ca alys (o amoun s acco ding o he ki ) we e placed.
• The diges ion was ca ied ou in a block a 375–420 °C un il he solu ion became clea (2–3 hou s depending
on he sample).
• A e cooling, he diges was ans e ed o he dis illa ion lask ( ill wi h wa e i necessa y).
• 40% NaOH was added o make he eac ion basic and elease NH₃.
• NH₃ was dis illed and cap u ed in a bo a e solu ion (o H₃BO₃).
• The solu ion con aining NH₄⁺ was i a ed wi h 0.1 N HCl un il he indica o colo changed.
• Pe o m he same s eps o he blank sample (diges wi hou sample) and s anda d (e.g. ammonium sul ama e)
o con ol.
2.6. Calcula ion
Ni ogen (mg) = (V _sample − V _blank) × N _HCl × 14.007 × 1000
• (V in L, N _HCl no mali y; 14.007 = a omic mass o N in mg/mmol)
• % Ni ogen = (mg N / sample mass (mg)) × 100
• % P o ein = % Ni ogen × con e sion ac o ( ypically 6.25)
• Example: i %N = 0.5 → %p o ein = 0.5 × 6.25 = 3.125%
The da a esul ing om he measu emen s we e s a is ically analyzed using ANOVA so wa e.
3. Resul s and discussion
Table 1.2. p esen s he esul s ela ed o he ca bohyd a e con en eco ded in he di e en basil geno ypes du ing he
3 yea s o s udy. They demons a e ha he highes alues o he a e age o he 3 yea s o s udy we e o he A oma ic
basil o Buzău a ie y 2.62 (g/100g) ollowed by he G eek Basil geno ype wi h alues o 2.07 (g/100g), ollowed by he
A ican Basil L8 geno ype wi h alues o 2.05 (g/100g). The lowes alues we e eco ded o he Siam Queen Basil
geno ype wi h alues o 1.23 (g/100g), ollowed by he Pe sian Basil geno ype wi h alues o 1.32 (g/100g). Bu as can
be seen om he a e age o he s udy yea s and pe plan , i is obse ed ha he weakes yea wi h low ca bohyd a e
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1102-1110
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con en alues was 2022, which leads o he conclusion ha en i onmen al condi ions again cons i u e a limi ing ac o
in e ms o ca bohyd a e con en .
(su as/sou ce: o iginal)
Figu e 1 Ca bohyd a e con en - wo k s ages
Table 2 Uni ac o ial analysis ega ding he ca bohyd a e con en o basil geno ypes analyzed in he h ee yea s o
expe ience (g/100g) (su sa/sou ce: o iginal)
N . Ca .
Geno ypes s udied
A e age ca bohyd a e con en
(g/100g)
Repo
Medigen
%
Di e ence
Medigen
Es ima ion
1.
Geno ypes s udied
2.63
145.68
0.82
* * *
2.
A oma ic Basil om
Buzau
1.88
104.10
0.07
3.
Pu ple basil Se a im
1.98
109.92
0.18
4.
Swee basil L1
1.89
104.65
0.08
5.
Lemon basil L2
1.83
101.33
0.02
6.
G eek basil
1.58
87.60
-0.22
7.
Basil wi h salad
lea es L4
1.63
90.38
-0.17
8.
G and Ve Basil L5
1.32
73.19
-0.48
o
9.
Pe sian basil
2.06
114.08
0.25
10.
Spicy Globe Basil L7
2.05
113.80
0.25
11.
A ican basil L8
1.87
103.54
0.06
12.
Holy basil a .
K ishna L9
1.84
102.02
0.04
13.
Macedonian basil
1.56
86.36
-0.24
14.
Dwa basil Sma ald
1.86
103.27
0.06
15.
Da k Opal ed basil
1.24
68.61
-0.56
o o
16.
Siam Queen Basil
1.65
91.48
-0.15
3.1. O e all a e age = 1.80
The uni ac o ial analysis ega ding he ca bohyd a e con en o he basil geno ypes s udied in he h ee yea s o
expe ience, wi h he so wa e ( he Ano a s a is ical p og am), in able 1.2., demons a ed ha he mos signi ican
posi i e di e ences a e ound in he A oma ic Basil o Buzău geno ype o 0.82, ollowed by Swee Basil L1 wi h 0.18 and
Spicy Globe Basil L7 wi h 0.25, which had he same esul as A ican Basil L8.
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Table 3 Uni ac o ial analysis ega ding he ca bohyd a e con en o basil geno ypes analyzed in he h ee yea s o
expe ience (g/100g) (su as/sou ce: o iginal)
SOURCE OF THE
VARIANT
VARIANT
GRADE o
LIBERTy
CORRECTED
DISPERSION
FACTOR
FISHER
CRITICAL FACTOR
(SPA)
(L)
(S2)
(F calcul)
10 %
5 %
1 %
0.1 %
BETWEEN
GROUPS
(SYSTEMATICS)
6.31
15
0.4207
19.3000
1.6480
1.9000
2.4800
3.3100
WITHIN GROUPS
(RESIDUAL)
1.05
48
0.0218
TOTAL
7.36
63
DIFERENCE
LIMIT (DL)
0.3500
0.4193
0.5592
0.7300
Using he Ano a so wa e, which is an analysis ool, he esul s in able 1.2. we e analyzed and we can see in able 1.3.,
om a s a is ical poin o iew, we can di e en ia e he in luence o he i s ac o , he in luence o he second ac o ,
as well as he combined in luence and he in e ac ion o he wo ac o s, whe e he Limi Di e ence (LD) a 5% is
0.4193%, a 1% is 0.5592% and a 1% is 0.7300%.
(su as/sou ce: o iginal)
Figu e 2 Uni ac o ial analysis ega ding he ca bohyd a e con en o basil geno ypes analyzed in he h ee yea s o
expe ience (g/100g)
The A ma ic Basil geno ype o Buzău showed a posi i e di e ence o 0.82, a alue ha is much highe han he DL (limi
di e ence) a 5% = 0.4193%, which means ha he di e ence is s a is ically signi ican . Consequen ly, his geno ype
showed a signi ican ly highe ca bohyd a e con en han he a e age o he geno ypes analyzed. Fo he Swee Basil L1
geno ype, he di e ence was 0.18 – lowe han he DL, so i is no s a is ically signi ican . The Spicy Globe Basil L7 and
A ican Basil L8 geno ypes wi h di e ences o 0.25 we e also eco ded below he signi icance h eshold o 0.4193%, so
hey a e also no s a is ically signi ican . The DL (Limi Di e ence) a 5% = 0.4193% cons i u es he h eshold below
which he di e ences be ween geno ypes a e no conside ed signi ican a a con idence le el o 95%. Only alues
exceeding his limi can be in e p e ed as eal and signi ican di e ences be ween geno ypes.
În conclusi e doa geno ype de Basil A oma ic o Buzău a p ezen a o di e ență semni ica i pozi i ă în ceea ce p i eș e
conținu ul de ca bohid ați, con o m analizei ANOVA. Celelal e geno ipu i (L1, L7, L8), deși au mani es a di e ențe
pozi i e, aces ea nu au os su icien de ma i pen u a i semni ica i e s a is ic. Aceas ă analiză sp ijină alege ea
geno ipului Basil A oma ic o Buzău pen u cul u i o ien a e că e un conținu idica de ca bohid ați.(g a ic 1.2.)
The able below ( able 1.4.) summa izes da a on he p o ein con en o basil geno ypes esea ched du ing 2021-2023.
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Rega ding p o ein con en , hei alues we e men ioned in able 1. 4. The highes alues o his p o ein con en we e
eco ded in he A oma ic o Buzău geno ype wi h he highes alue in 2021 wi h 3.29 (g/100g), ollowed by 2022 wi h
a alue o 3.22 (g/100g) and wi h an a e age alue o he s udy yea s o 3.10 (g/100g). Ano he geno ype wi h high
p o ein con en alues was Se a im Viole Basil, which eco ded a e age alues o 2.41 (g/100g) ollowed by he A ican
Basil L8 geno ype wi h a e age p o ein con en alues o 1.90 (g/100g). The lowes p o ein alues we e eco ded o
he Swee Basil L1 geno ype wi h an a e age alue o 1.09 (g/100g), ollowed by he Siam Queen Basil geno ype wi h an
a e age alue o 1.22 (g/100g). The p o ein con en alues we e in luenced by he geno ype and he en i onmen al
condi ions o he yea s in which he esea ch was conduc ed.
F om a s a is ical poin o iew, using he Ano a so wa e, he di e ence in he a e age alue o he p o ein con en
(g/100g) is signi ican ly posi i e o he A oma ic Basil o Buzau, compa ed o he Swee Basil L1, acco ding o able 1.4.
The A oma ic o Buzău geno ype eco ded he highes p o ein con en , and i s alues signi ican ly exceeded he DL,
which con i ms ha he di e ences om o he geno ypes a e s a is ically signi ican . Pu ple Basil Se a im and A ican
Basil L8 eco ded an a e age p o ein con en ha was p obably signi ican compa ed o hose wi h e y low alues.
Swee Basil L1 and Siam Queen we e a he lowe end, wi h subs an ially lowe alues, and he di e ences om he
highe geno ypes exceeded he DL, so hey a e signi ican .
Table 4 Resul s ega ding he p o ein con en (g/100g) o he basil geno ypes analyzed in he h ee yea s o expe ience
N ca .
Geno ypes
s udied
P o ein con en
(g/100g)/2021
P o ein con en
(g/100g)/2022
P o ein con en
(g/100g)/2023
A e age p o ein
con en alues pe
geno ype/3 yea s o
s udy (g/100g
)
1.
A oma ic
Basil om
Buzau
3,29
3,02
2,99
3.10
2.
Pu ple basil
Se a im
2,58
2,42
2,25
2.42
3.
Swee basil
L1
1,14
1,01
1,12
1.09
4.
Lemon basil
L2
1,59
1,42
1,51
1.51
5.
G eek basil
1,74
1,63
1,64
1.67
6.
Basil wi h
salad lea es
L4
1,92
1,85
1,36
1.71
7.
G and Ve
Basil L5
1,45
1,32
1,31
1.36
8.
Pe sian
basil
1,55
1,32
1,42
1.43
9.
Spicy Globe
Basil L7
1,79
1,65
1,56
1.67
10.
A ican
basil L8
1,92
1,95
1,85
1.91
11.
Holy basil
a . K ishna
L9
1,75
1,67
1,52
1.65
12.
Macedonian
basil
1,65
1,61
1,52
1.59
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13.
Dwa basil
Sma ald
1,91
1,47
1,68
1.69
14.
Da k Opal
ed basil
1,75
1,63
1,51
1.63
15.
Siam Queen
Basil
1,22
1,32
1,12
1.22
16.
Basil
cinnamon
1,37
1,31
1,25
1.31
(su as/sou ce: o iginal)
The uni ac o ial ANOVA analysis o p o ein con en o he basil geno ypes s udied in he pe iod 2021–2023 e ealed
s a is ically signi ican di e ences. The A oma ic basil o Buzău geno ype eco ded he highes a e age alue (3.10
g/100g), wi h a signi ican di e ence compa ed o he o he geno ypes, exceeding DL = 0.3320%. In con as , he Swee
Basil L1 (1.09 g/100g) and Siam Queen (1.22 g/100g) geno ypes eco ded he lowes alues, wi h signi ican nega i e
di e ences compa ed o he o he geno ypes.
Table 5 Resul s ega ding he p o ein con en (g/100g) o he basil. geno ypes analyzed in he h ee yea s o expe ience
N . Ca .
Geno ypes s udied
A e age p o ein con en (g/100g)
Repo
MedGen %
Di e ence
MedGen
Es ima ion
1.
A oma ic Basil om
Buzau
3.10
183.55
1.41
* * *
2.
Pu ple basil Se a im
2.42
143.14
0.73
* * *
3.
Swee basil L1
1.09
64.54
-0.59
o o o
4.
Lemon basil L2
1.51
89.26
-0.18
5.
G eek basil
1.67
98.88
-0.01
6.
Basil wi h salad
lea es L4
1.71
101.25
0.02
7.
G and Ve Basil L5
1.36
80.53
-0.32
8.
Pe sian basil
1.43
84.67
-0.25
9.
Spicy Globe Basil L7
1.67
98.73
-0.02
10.
A ican basil L8
1.91
112.94
0.22
11.
Holy basil a .
K ishna L9
1.72
101.99
0.03
12.
Macedonian basil
1.59
94.29
-0.09
13.
Dwa basil Ema ald
1.69
99.92
0.00
14.
Da k Opal ed basil
1.63
96.51
-0.05
15.
Siam Queen Basil
1.22
72.24
-0.46
o o
16.
Basil cinnamon
1.31
77.56
-0.37
o
(su as/sou ce: o iginal)
The one-way analysis (Ano a) on he p o ein con en o he 16 basil geno ypes is summa ized in Table 1.6.
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Table 6 Resul s ega ding he p o ein con en (g/100g) o he basil geno ypes analyzed in he h ee yea s o expe ience
SOURCE OF THE
VARIANT
VARIANT
GRADE o
LIBERTY
CORRECTED
DISPERSION
FACTOR
FISHER
CRITICAl FACTOR
(SPA)
(L)
(S2)
(F calcula )
10 %
5 %
1 %
0.1 %
BETWEEN
GROUPS
(SYSTEMATICS)
14.06
15
0.9374
68.5976
1.6480
1.9000
2.4800
3.3100
IN INTERIORUL
GRUPELOR
(REZIDUALA)
0.66
48
0.0137
TOTAL
14.72
63
DIFERENCE
LIMIT (DL)
0.2771
0.3320
0.4427
0.5780
(su as/sou ce: o iginal)
Table 6 summa izes da a on he suga con en o he 16 basil geno ypes unde esea ch in he pe iod 2021-2023.
(su as/sou ce: o iginal)
Figu e 3 Uni ac o ial analysis o p o ein con en (g/100g) o basil geno ypes analyzed in he h ee yea s o expe ience
4. Conclusion
In conclusion, only he A oma ic o Buzău Basil geno ype showed a signi ican posi i e di e ence in e ms o
ca bohyd a e con en , acco ding o he ANOVA analysis. The o he geno ypes (L1, L7, L8), al hough hey showed
posi i e di e ences, hese we e no la ge enough o be s a is ically signi ican . This analysis suppo s he choice o he
A oma ic o Buzău Basil geno ype o c ops o ien ed owa ds a high ca bohyd a e con en .
The p o ein con en alues we e in luenced by he geno ype and he en i onmen al condi ions in he yea s in which he
esea ch was ca ied ou . The A oma ic basil o Buzău geno ype eco ded he highes p o ein con en , and i s alues
signi ican ly exceeded he DL, which con i ms ha he di e ences compa ed o o he geno ypes a e s a is ically
signi ican . Se a im Viole Basil and A ican Basil L8 eco ded an a e age p o ein con en ha was p obably signi ican
compa ed o hose wi h e y low alues. Swee basil L1 and Siam Queen we e loca ed a he lowe end, wi h subs an ially
lowe alues, and he di e ences om he highe geno ypes exceed he DL, so hey a e signi ican .
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), 1102-1110
1110
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