© 2022 Jou nal o Di e si y S udies. All Righ s Rese ed.
Compa a i e Phy ochemical Analysis o Hemi Pa asi e Lo an hus
longi lo us Des ; on Hos Azadi ac a indica by GCMS Analysis
Ch. Uday Kuma *, Lakshmi Bha ani
Depa men o Bo any, Uni e si y College o Science, Sai abad Campus, Osmania Uni e si y, Hyde abad-India
INTRODUCTION
Some plan s acqui e hei ood o wa e om
o he plan s, which a e known as pa asi es,
and he sou ce p o iding plan s a e known as
hos s. The s em pa asi ic plan s a e commonly
named mis le oes. Mis le oes a e de i ed om
he Anglo-Saxon wo d mis elian which means,”
mis ” o dung and “ an” o wig [1]. Lo an hus
longi lo us Des . commonly known as Mis le oe,
all-heal, de ils uge, belongs o Lo an haceae
amily wi h 76 Gene a and 1000 species. The
synonym o Lo an hus is Dend oph hoe alca a.
The semi-pa asi es possess lea es and syn hesize
he ca bohyd a es po ion o hei ood. These
pa asi es es ablish ela ions wi h hos ascula
elemen s o d aw nu ien s [2-3]. Hemi pa asi es
ha e a pa asi ic ela ionship wi h mo e han 300
hos plan s [4]. Unlike o he pa asi es, Lo an hus
seeds do no equi e a ge mina ion s imulan om
he li ing hos [5]. Nume ous hemi pa asi es do no
syn hesize o modi y he seconda y compounds
aken up om hei hos s [6], hus he p esence o
ce ain seconda y compounds a ies wi hin and
among he popula ions o hemi pa asi ic plan s,
depending on he hos associa ion o indi idual
pa asi es [7], i has been used in adi ional
medicine and ound o ha e an imic obial, an i
diabe ic, an ioxidan , an icance , hype ensi e
and an i i al p ope ies [8-10]. They a e used as
a heal h ood o enhancing immuni y and used as
a pain elie e , aph odisiac, na co ic and diu e ic
[11-12]. The aim o he s udy was compa a i e
analysis o hos -Azadi ach a indica and pa asi e-
Lo an hus longi lo us o phy ochemicals.
P epa a ion o plan ma e ial powde
The esh lea es o pa asi e-Lo anhus longi lo us
Abs ac
Plan pa asi es a e known o ob ain p ima y and seconda y compounds om hei hos s. Mis le oes o pa asi ic plan s
a e g owing on a ious ees and belong o he Lo an haceae amily. Nume ous mis le oes don’ syn hesize o modi y
he seconda y compounds aken up om hei hos s’.The hos s signi ican ly in luenced he phy ocons i uen s o hemi
pa asi es. In adi ional sys ems as well as in he mode n sys em o medicine he plan Azadi ach a indicais used as a
medicine o a ious ailmen s and so he pa asi e Lo an hus longi lo us g own on neem ee was chosen o he s udy.
The p esen s udy is ca ied ou o de ec he phy ochemical cons i uen s o Hos -Azadi ach a indica and pa asi e
Lo an hus longi lo us om he Hyde abad egion. Phy ochemical analysis o me hanolic lea ex ac s e ealed he
p esence o annins, phenols, and An h aquinones in bo h he hos and pa asi e. GCMS analysis e ealed he p esence
o i e bioac i e compounds in he hos and pa asi e wi h a ied composi ions. The phy ochemical and GCMS analysis
indica ed simila i y in p esence o phy ocons i uen s in bo h he hos and pa asi e.
Keywo ds: GCMS, Lo an hus longi lo us, Plan pa asi es, phy ochemical cons i uen s
Ci a ion: Ch.Uday Kuma , Lakshmi Bha ani (2022). Compa a i e Phy ochemical Analysis o Hemi Pa asi e
Lo an hus longi lo us Des : on Hos Azadi ac a indica by GCMS Analysis. Jou nal o Di e si y S udies. V01i01, 01-04.
DOI: h p://dx.doi.o g/10.5281/zenodo.17656915
Co esponding Au ho : Ch. Uday Kuma
A icle His o y: Recei ed on: July 12, 2022 | Re ised on: Sep embe 12, 2022 | Accep ed on: Oc obe 10, 2022
Copy igh : © 2022 by he au ho s. The license o Jou nal o Di e si y S udies. This a icle is an open access
a icle dis ibu ed unde he e ms and condi ions o he C ea i e Commons A ibu ion (CC BY) license (h p://
c ea i ecommons o g/licenses/by/4.0/).
E-mail Add ess: [email p o ec ed]
ORIGIONAL RESEARCH ARTICLE
2© 2022 Jou nal o Di e si y S udies. All Righ s Rese ed.
Ch. Uday Kuma ., / Jou nal o Di e si y S udies (2022)
and hos -Azadi ach a indica we e collec ed om
Hyde abad. The plan ma e ial was washed
ho oughly i s wi h ap wa e and hen dis illed
wa e o emo e he su ace con aminan s.
La e hey a e shade d ied sepa a ely a oom
empe a u e (30°C±5°C) o abou wo weeks
o ge cons an weigh and hen pul e ized in o
a d y powde using an elec ic blende . Un il
needed, he powde ed lea samples we e kep in
an ai igh con aine .
P epa a ion o ex ac
The d ied plan ma e ials o L. longi lo us and
Azadi ach a indica lea es samples we e ex ac ed
wi h Me hanol in soxhle appa a us o 3h s. The
ex ac was cooled, il e ed, and concen a ed
using a acuum lask e apo a o . Finally, his
ex ac was dissol ed in 1ml me hanol and
cen i uged a 3000 pm o 5min.
Phy ochemical Analysis
Phy ochemical analysis o me hanol lea es ex ac
was ca ied ou ollowing ce ain p esc ibed
me hods adop ed by S ini as R. e al., 2014.
Tes o Tannins
In 2ml o dis illed wa e , a ew d ops o e ic
chlo ide solu ion we e added o oughly 2ml
o ex ac . The appea ance o g een- in ed
p ecipi a e con i ms he p esence o annins.
Tes o Saponins
In a es ube, 3 ml o he ex ac was mixed wi h
3ml o dis illed wa e and igo ously shaken o
a ew momen s. The es ube was hea ed, and
c ea ing s able oam indica es he p esence o
saponins.
Tes o Fla onoids
1ml o 10 pe cen lead ace a e solu ion was added
o 1 ml o ex ac in a es ube. The appea ance o
he yellow p ecipi a e can de e mine he p esence
o la onoids.
Tes o Alkaloids
On a ho wa e ba h, 3ml o he ex ac was mixed
wi h 3 ml o 1% HCl. Then 1ml o he liquid was
spli be ween wo es ubes.
D agend o ’s Tes
1 ml o D agend o ’s eagen was added o
he i s es ube. The p esence o o ange- ed
p ecipi a e was ega ded as a good sign.
Maye ’s Tes
1ml o Maye ’s eagen was added o he second
es ube. The o ma ion o a bu -colo ed
p ecipi a e can con i m he p esence o alkaloids
Tes o Te penoids
2 ml ex ac was dissol ed in 2 ml CHCl3 and
e apo a ed o d yness. A e ha , 2 ml o
concen a ed sulphu ic acid (H2SO4) was added
and hea ed o abou 2 minu es. The eme gence
o a g ayish colo indica es he p esence o
e penoids.
Tes o S e oids
Two es s we e ca ied ou o pe cei e he
p esence o s e oids in he ex ac , and hose we e
he Salkowski’s es and he Liebe mann es .
Salkowski’s es : 2ml o he o ganic ex ac was
dissol ed in 2ml o chlo o o m. 2ml concen a ed
H2So4 was added o his. The p esence o s e oids
is shown by he appea ance o ed colo in he
chlo o o m po ion.
Tes o Phenols
5ml dis illed wa e was added o 1gm o ex ac ,
and he solu ion was hen ea ed wi h a 5% e ic
chlo ide solu ion. The p esence o phenols is
con i med by o ming a da k g een colo .
GC-MS Analysis
The GC-MS analysis o heme hanolic lea es
ex ac was done a he Cen al Analy ical Facili y,
Uni e si y College o Technology, Osmania
Uni e si y using he s anda d GCMS model. The
pa asi e plan Lo an hus longi lo us and hos
Azadi ac a indica lea es ex ac s we e analyzed
using a GCMS-QP2010 SHIMADZU, Japan, used
wi h an Op ima 5 ms capilla y column (30 0.25
mm) 0.25 m ilm hickness, as desc ibed by
3© 2022 Jou nal o Di e si y S udies. All Righ s Rese ed.
Ch. Uday Kuma ., / Jou nal o Di e si y S udies (2022)
RESULTS AND DISCUSSION
The phy ochemical analysis o he me hanol
ex ac o wo plan lea es Lo an hus longi lo us
–pa asi e and hos -Neem e ealed he p esence
o annins, phenols, an h aqinones and he
absence o saponins and alkaloids. Fla onoids and
s e oids absen in he pa asi e a e p esen in hos
(Table-1 and Fig-3). The GC-MS ch oma og aphic
p o ile o he hos and pa asi e plan lea es
ex ac s showed a ious signi ican cons i uen s
like Di luo ine monoxide; Ni osyi chlo ide;
1-P opyne,3-chlo o; Py idazine, 3-me hoxy; N,N
Dini o 1,3,5,7 e azabicyclononane in di e se
composi ions. Hos plan con ains he compounds
o 2-Nonynoic acid (15.26%), Me hane, chlo o
T ini on (13.04%) ha we e absen in pa asi e
plan ollowed by Di luo ine monoxide (11.28%),
Ni osyi chlo ide (2.11%), 1-P opyne, 3-chlo o-
(0.60%) N,N Dini o1,3,5,7 e azabicyclononane
(0.41%), Py idazine, 3-me hoxy (0.28%). Pa asi ic
plan con ains highes le el o Phenyle hylene.3-
me hoxy-2,2-dini o (62.40%) ollowed by
Di luo ine monoxide (0.40%), Ni osyi chlo ide
(0.38%), Py idazine, 3-me hoxy (0.13%),
1-P opyne, 3-chlo o-(0.13%), N, N Dini o1, 3, 5, 7
e azabicyclononane (0.34%). 2-Nonynoic acid,
Me hane, chlo o ini on ha we e absen in hos
plan s a e absen inpa asi e plan s.
Fig 1: Ch oma og am Neem Pa asi e E:/2022/
OCT/10-uday-GCMS/Neem pa asi e.
Fig 2: Ch oma og am Neem Hos E:/2022/
OCT/10-uday-GCMS/Neem Hos .
Table 1: Phy ochemical analysis o pa asi e-L.
longi lo us and Hos - Azadi ac a indica
S
no Cons i uen s Hos - Azadi ac-
a indica
Pa si e- L.longi-
lo us
1Tes o Alkaloids Absen Absen
2 Tes o annins P esen P esen
3 Tes o saponins Absen Absen
4Tes o la onoids P esen Absen
5Tes o e penoids Absen P esen
6 Tes o s e oids P esen Absen
7 Tes o Phenols P esen P esen
8Tes o An h a-
quinones P esen P esen
Fig 3: Phy ochemical analysis o Hos and Pa asi e
plan s.
(A)Tannins (B) Saponins (C) Fla onoids (D)
Alkaloids (E) Te penoids (F) S e oids (G) Phenol
(H) An h oquinones
Iheagwame al (2019). The ini ial column o en
empe a u e (60°C) is planned o climb o 160°C
a a a e o 10°C/min, hen o 250°C wi h hold
du a ion o 2 min/inc emen . An injec ion olume
o 1.0 L in he spli less mode wi h a spli a io o
1:1 and injec o empe a u e se a 200° C wi h
mino adjus men s. The ollowing we e he mass
spec opho ome e se ings: 230°C o he ion
sou ce, 250°C o he in e ace, 4.5 minu es o he
sol en delay, and 50–700 amu o he scan ange.
The mul iplie ol age and elec on ioniza ion
mode we e modi ied o 70 eV and 1859 V. Fo
compound iden i ica ion, he e en ion ime,
agmen a ion pa e n, and mass spec al da a
o unknown componen s in he ex ac s equa ed
o hose in he Wiley and Na ional Ins i u e o
S anda ds and Technology (NIST) lib a ies [14-
24].
4© 2022 Jou nal o Di e si y S udies. All Righ s Rese ed.
Ch. Uday Kuma ., / Jou nal o Di e si y S udies (2022)
CONCLUSION
Hos and pa asi e plan lea es ex ac s depic s
simila Bioac i e compounds like Di luo ine
monoxide, Ni osyi chlo ide, 1-P opyne,3-chlo o,
Py idazine, 3-me hoxy, N,N Dini o 1,3,5,7
e azabicyclononane wi h a ied composi ion.
The analysis can be used as a use ul ool o
pha macognacy. Finally, he p esen epo s poin s
owa ds he need o u he in es iga ions on he
possible ole o ans e ed seconda y compounds
ei he as media o s o hos de ense, hos de ense
compounds u ilize as cues o iden i ica ion o
hos by he hemi pa asi e o compounds aken up
by he pa asi e o suppo hei de ense agains
ejec ion by he hos .
Acknowledgmen
Au ho s a e hank ul o he Depa men o
Bo any, UCSS Sai abad and Uni e si y College o
Technology, Osmania Uni e si y, Hyde abad and
Telangana o hei aluable help du ing he ime
o Ph.D. wo k
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Table 2: Phy o compounds iden i ied by GC-MS analysis.
S.
NO
COMPOUND
NAME
RETENTION
TIME MOLICULAR
WEIGHT FORMULA
CAS
NO %AREA
HOST
%AREA
PARSITE
(H.N) (P.L)
1Di luo ine monoxide 0.108 3.060 54 F2O 7783-41-7 11.28% 0.40%
2 Ni osyi chlo ide 0.595 3.875 65 ClNO 2696-92-6 2.11% 0.38%
3 1-P opyne,3-chlo o- 2.180 3.505 74 C3H3Cl 624-65-7 0.60% 0.13%
4Py idazine, 3-me hoxy 2.180 3.505 110 C6H6N2O19064-
65-4 0.28% 0.13%
5N,N Dini o 1,3,5,7
e azabicyclononane 2.545 3.584 218 C5H10N6O 0-00-0 0.41% 0.34%
6 2-Nonynoic acid 0.059 0 154 C9H14O21846-70-4 15.26% 0
7Phenyle hylene,3-me-
hoxy-2,2-dini o 0 2.429 224 C9H8N2O516990-0-2 0 62.40%
8 Me hane, chlo o ini o 0.200 0 185 CClN3O61943-16-4 13.04% 0
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