Metabolite Profiling of 96 % Ethanol Extract

Marsilea crenata Presl. Leaves using UPLC-QTOF-MS/MS

Agnis Pondinekaria Aditama

1

*, Mangestuti Agil

2

1

Doctoral Program of Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia

2

Department of Pharmacognocy and Phytochemistry, Faculty of Pharmacy,

Universitas Airlangga, Surabaya, Indonesia

Abstract: Background: Marsilea crenata Presl. plants grow in east java area, usually consumed by local people,

and was known having medical purposes. Some researches were conducted toward to the plant and

showed that the plant having potential treatment to some diseases. Objective: The aim of research is to

know the contain of Marsilea crenata Presl. compound by using UPLC MS/MS methode. Methods:

Marsilea crenata Presl. M crenata was ekstracted using etanol 96% by using Ustrasonic Assisted

Extraction methode. The first step was prepare 100 extrac ppm, and then were injected 5 µL to UPLC

MS/MS. The next step, the data obtained was total ion chromatogram (TIC), and the last step, data was

analyzed by using soffware Masslynx 4.1. Results: Which shown in each equipment dichloromethane

(DCM) blank 47 compound and methanol blank 50 compound. Conclusion: This is the first report of the

application of non-targeted metabolomics in Marsilea crenata Presl.

Keywords: Marsilea crenata Presl., metabolite profiling, UPLC-QTOF-MS/MS, 96% ethanol.

1 INTRODUCTION

Marsilea crenata Presl. Contains of different

phytochemical which having medical purposes. Tthe

benefit explained above is the effect of metabolit

secunder that was obtained in Marsilea crenata

Presl. Secondary metabolism is chemical material

that was resulted from the plant metabolism process

that is useful to the plant. Secondary metabolism is

classified according to chemical structured

functional characteristic such as alkaloid, flavonoid,

saponin, tannin, poliphenole, antraquinone and

volatile oil (Manitto, 1992; Jacoeb et al., 2010).

Some research had been done to know the activity of

Marsilea crenata Presl. Some of them are, Marsilea

crenata Presl. Leaves had been observed by using

Radio Immuno Assay (RIA) and activity observation

in vivo in female mice. The result showed that 96 %

ethanol extract Marsilea crenata Presl. Leaves

enable to inhibit osteoporosis to pascamenopouse

woman by increasing bone remodelling process

mechanism especially in the bone forming (Putra

and Laswati, 2011).

The research that had been done was Gas

Chromatography-Mass Spectrometry (GC-MS)

analysis where the result showed that some

compound such as monoterpenoid, diterpenoid, fatty

acid, and other compound have not been known in

n-hexane extract of Marsilea crenata Presl. Leaves.

and Palmitat contain was assumed enable to increase

the bone forming process with induction mechanism

in osteoblast cell so that it can be used as

phytoestrogen (Ma’arif et al., 2016).

According to the previous research GC-MS

instrument was used in order to know Marsilea

crenata Presl. Metabolit secondary contain, but not

all secondary metabolit chemical compound can be

analysed because lack of instrument, so only volatile

compound can be analysed. Periodic and update

library is needed because there are some compound

having similar m/z model, so it is known as

similarity index (SI). Therefore metabolit profiling

must be done by using Ultra Performance Liquid

Chromatography-Mass Spectrometer (UPLC-MS)

instrument. UPLC-MS instrument is liquid

chromatography technique with mass spectrometer

detector. Bio analysis research use UPLC-MS. The

instrument is specific and having wide application as

well as practical method. The application of this

instrument is not restricted only for volatile

molecule, high flecsibility and limited time (K

Naresh et al., 2014; Chawla and Ranjan, 2016). The

Metabolite Proﬁling of 96 .

DOI: 10.5220/0008357000210035

In Proceedings of BROMO Conference (BROMO 2018), pages 21-35

ISBN: 978-989-758-347-6

Copyright

c

21

using of UPLC-MS can give scientific data that is

benefical for the user of the plant drug.

2 MATERIAL AND METHOD

2.1 Material

We performed UPLC-QTOF-MS/MS (Waters),

Oasis C18 Cartridge (Waters), Sonicator (Sonica),

Moisture Analyzer (Mettler Toledo), Vacuum

Rotary Evaporator (Heidolph), TLC (Camag), TLC

Visualizer (Camag), analytical scales (XX), flasks,

beaker glass, measuring cups, petri dishes, stirrer

bars, spatulas, dropper pipes, funnel, filter paper,

eppendorf, and computers.

Marsilea crenata Presl. Leaves were obtained

from Benowo village at Surabaya, ethanol 96%

(Merck), aquadest, dichloromethane (Merck),

acetonitrile (Merck) and formic acid (Merck).

2.2 Methods

2.2.1 Sample Preparation

The extract preparation was done by simplicia of

Marsilea crenata Presl. Leaves powder weighed

30 g and put into the Erlenmeyer flask, then

dissolved with 500 ml ethanol solvent with

replication 3 times (200 ml, 150 ml, 150 ml). Further

extraction is done with the help of ultrasonic waves

(> 20 kHz) for 6 minutes with 3 pauses every 2

minutes. Ethanol 96% extraction was performed by

single extraction. The extract was evaporated using a

Rotary evaporator, then stored in an oven with a

temperature of 40

0

C.

2. Extract Preparation to UPLC-QTOF-MS/MS

Analysis

Sample was injected to instrument UPLC

MS/MS 5µl, and than chromathogram was obtained

and the data was processed by using software

Masslynx so that peak area, retention time, spectra

m/z dan elemental composition was obtained from

each peak area was detected. The next step, data

interpretation was done by using website

Chemspider to get the level of data similarity from

chromagram and spectra, so that the similarity

explained above, we can get the suitable IUPAC

name and it can be concluded that metabolit contain

was in M.crenata extract.

3 RESULT AND DISCUSSION

The extraction method used by ultrasonic assisted

extraction (UAE) which has advantages, among

others, accelerating the extraction process

(compared with conventional extraction eg

maceration), more time efficient, and can increase

the crude rendement rate of the extract. In addition,

ultrasonic extraction may also be used in the

extraction of heat resistant materials (Handayani et

al., 2016).

Fourty seven compounds in DCM blank

and Fifty compounds in methanol blank were

obtained from UPLC MS/MS analysis. Data

obtained was total ion of kromatogram (TIC) and 96

% ethanol extract from Marsilea crenata Presl.

leaves that was processed by using software

Masslynx so that peak area, retention time, spectra

m/z dan elemental composition was obtained from

each peak area was detected. The next step, data

interpretation was done by using website

Chemspider to get the level of data similarity from

chromagram and spectra, so that the similarity

explained above, we can get the suitable IUPAC

name.

Fifty major contain were tentatively assigned

based on their accurate masses, MS/MS

fragmentation patterns in methanol blank and Forty-

seven major contain in dichloromethane (DCM)

blank, in comparison to standard compounds and

references (Table 1 and 2).

Table 1: Metabolite profiling Marsilea crenata Presl.in methanol blank by UPLC-QTOF-MS/MS

No.

RT

% Area

formula

Trivial name

IUPAC name

Activity

1

0,200694

0,0039%

-

2

0,478472

0,0014%

-

22

3

1.535

2,4313%

C10H21NO5

4-(3-

Hydroxypropyl

)-4-nitro-1,7-

heptanediol

4-(3-

Hydroxypropyl)-

4-nitro-1,7-

heptanediol

-

4

2.232

0,1510%

C11H21NO7

2-[(tert-

Butoxycarbony

l)amino]-2-

deoxy-D-

glucopyranose

2-Deoxy-2-({[(2-

methyl-2-

propanyl)oxy]car

bonyl}amino)-D-

glucopyranose

-

5

2.518

1,5144%

C12H23NO7

Methyl 4,6-

dideoxy-4-

{[(2R)-2,4-

dihydroxybuta

noyl]amino}-2-

O-methyl-α-D-

mannopyranosi

de

Methyl 4,6-

dideoxy-4-

{[(2R)-2,4-

dihydroxybutano

yl]amino}-2-O-

methyl-α-D-

mannopyranoside

-

6

3.799

1,4856%

C15H21NO7

Methyl (3,4,5-

triethoxy-2-

nitrophenyl)ace

tate

Methyl (3,4,5-

triethoxy-2-

nitrophenyl)acetat

e

-

7

4.427

1,4055%

C5H15N3Cl2

4-

Hydrazinopiper

idine

dihydrochlorid

e

4-

Hydrazinopiperid

ine

dihydrochloride

-

4.610

0,3629%

C9H6O3

3

hydroxycouma

rin

3-Hydroxy-2H-

chromen-2-one

Penghambatan

kompetitif

DAAO

rekombinan

manusia (Molla,

2017).

8

4.896

0,1836%

C20H24N3S

Cl

Prochlorperazi

ne

2-Chloro-10-[3-

(4-methyl-1-

piperazinyl)propy

l]-10H-

phenothiazine

Analgesik

(callan, 2008),

amtiemetik

(roberge, 2006)

9

5.228

0,9215%

C13H18N5O

5Cl

Ethyl 4-[3-(4-

chloro-3-nitro-

1H-pyrazol-1-

yl)propanoyl]-

1-

piperazinecarb

oxylate

Ethyl 4-[3-(4-

chloro-3-nitro-

1H-pyrazol-1-

yl)propanoyl]-1-

piperazinecarbox

ylate

-

10

5.445

0,0257%

C33H37N3

4-{Bis[4-(1-

pyrrolidinyl)ph

enyl]methyl}-

N,N-dimethyl-

4-{Bis[4-(1-

pyrrolidinyl)phen

yl]methyl}-N,N-

dimethyl-1-

-

Metabolite Proﬁling of 96

23

1-

naphthalenami

ne

naphthalenamine

11

5.628

0,9906%

C10H21N3O

8S

1-Azido-1-

deoxy-2,3-bis-

O-

(methoxymeth

yl)-5-O-

(methylsulfony

l)-D-ribitol

1-Azido-1-deoxy-

2,3-bis-O-

(methoxymethyl)-

5-O-

(methylsulfonyl)-

D-ribitol

-

12

5.845

0,6908%

C29H18N4O

6S

2-(2-{(E)-2-

Cyano-2-[4-(2-

oxo-2H-

chromen-3-yl)-

1,3-thiazol-2-

yl]vinyl}-4-

nitrophenoxy)-

N-

phenylacetami

de

2-(2-{(E)-2-

Cyano-2-[4-(2-

oxo-2H-chromen-

3-yl)-1,3-thiazol-

2-yl]vinyl}-4-

nitrophenoxy)-N-

phenylacetamide

-

13

6.177

1,0895%

C25H22O11

4-(1,3-

Benzodioxol-5-

yl)-6-hydroxy-

1-oxo-1,3-

dihydronaphth

o[2,3-c]furan-

5-yl

hexopyranosid

e

4-(1,3-

Benzodioxol-5-

yl)-6-hydroxy-1-

oxo-1,3-

dihydronaphtho[2

,3-c]furan-5-

yl hexopyranosid

e

-

14

6.577

0,3205%

C24H22O14

2-(3,4-

Dihydroxyphen

yl)-5-hydroxy-

4-oxo-4H-

chromen-7-yl

6-O-

(carboxyacetyl)

-β-D-

glucopyranosid

e

2-(3,4-

Dihydroxyphenyl

)-5-hydroxy-4-

oxo-4H-chromen-

7-yl 6-O-

(carboxyacetyl)-

β-D-

glucopyranoside

-

15

6.908

0,2713%

C14H21NO

1-[1-(4-

Methoxypheny

l)cyclohexyl]m

ethanamine

1-[1-(4-

Methoxyphenyl)c

yclohexyl]methan

amine

-

24

16

7.206

2,0878%

C11H16O3

1-carboxy-3-

hydroxyadama

ntane

3-Hydroxy-1-

adamantanecarbo

xylic acid

-

17

7.423

0,6567%

C16H23NO2

UNII:891H89

GFT4

1-(7-Ethyl-1-

benzofuran-2-yl)-

2-[(2-methyl-2-

propanyl)amino]e

thanol

-

18

7.640

0,2325%

C11H24N5Cl

1-Hexyl-6,6-

dimethyl-1,6-

dihydro-1,3,5-

triazine-2,4-

diamine hydroc

hloride (1:1)

1-Hexyl-6,6-

dimethyl-1,6-

dihydro-1,3,5-

triazine-2,4-

diamine

hydrochloride

(1:1)

-

19

7.903

0,3096%

C14H22N5Cl

1-methyl-2-[(4-

methylpiperazi

n-1-

yl)methyl]benz

imidazol-5-

amine

hydrochloride

1-Methyl-2-[(4-

methyl-1-

piperazinyl)meth

yl]-1H-

benzimidazol-5-

amine

hydrochloride

(1:1)

-

20

8.406

1,4141%

C36H46N4O

Manzamine J

(1R,2R,12R,13S,

16Z)-25-(9H-β-

Carbolin-1-yl)-

11,22-

diazatetracyclo[1

1.11.2.12,22.02,1

2]heptacosa-

5,16,25-trien-13-

ol

-

21

8.886

0,0560%

C17H31NO9

6-O-(N-{[(2-

Methyl-2-

propanyl)oxy]c

arbonyl}-D-

leucyl)-α-D-

allopyranose

6-O-(N-{[(2-

Methyl-2-

propanyl)oxy]car

bonyl}-D-leucyl)-

α-D-allopyranose

-

22

9.321

0,1071%

C18H27NO2

dyclonine

1-(4-

Butoxyphenyl)-3-

(1-piperidinyl)-1-

propanone

Inhibitor

Aldehyde

Dehydrogenase

1 (ALDH1A1)

(Collard, 2007).

Antimicroba

(Florestano,1956

)

Metabolite Proﬁling of 96

25

23

9.584

0,1649%

C13H29N3O

4S

(3R,4R)-3-

{[(2-

Hydroxyethyl)(

methyl)amino]

methyl}-4-

(hydroxymethy

l)-N-isopropyl-

N-methyl-1-

pyrrolidinesulf

onamide

(3R,4R)-3-{[(2-

Hydroxyethyl)(m

ethyl)amino]meth

yl}-4-

(hydroxymethyl)-

N-isopropyl-N-

methyl-1-

pyrrolidinesulfon

amide

-

24

10.601

0,6568%

C12H18NO

N,N,N-

Trimethyl-3-

oxo-3-phenyl-

1-

propanaminiu

m

N,N,N-

Trimethyl-3-oxo-

3-phenyl-1-

propanaminium

-

25

10.830

0,3341%

C47H61N3O

8S

2-

({(3β,7β,8ξ,9ξ,

10α,12β,13α,1

4ξ,17α,20S)-3-

[(2-{[(3-

Acetyl-2-

methyl-4-

quinolinyl)ami

no]methyl}phe

nyl)ethynyl]-

3,7,12-

trihydroxy-24-

oxocholan-24-

yl}amino)ethan

esulfonic acid

2-

({(3β,7β,8ξ,9ξ,10

α,12β,13α,14ξ,17

α,20S)-3-[(2-

{[(3-Acetyl-2-

methyl-4-

quinolinyl)amino]

methyl}phenyl)et

hynyl]-3,7,12-

trihydroxy-24-

oxocholan-24-

yl}amino)ethanes

ulfonic acid

-

26

11.082

0,4582%

-

27

11.379

0,8714%

C37H47N9O

S

-

28

11.562

1,7782%

C14H19N4O

2Cl

Lintopride

4-Amino-5-

chloro-N-[(1-

ethyl-4,5-

dihydro-1H-

imidazol-2-

yl)methyl]-2-

methoxybenzami

de

-

29

11.928

0,4325%

C28H49NO1

2-Methyl-2-

propanyl 2-

2-Methyl-2-

propanyl 2-

-

26

2

cyano-3-

[(4S,5R)-5-

{(5S,6R)-6-

[(4R)-2,2-

dimethyl-1,3-

dioxolan-4-yl]-

2,4,7,9-

tetraoxadecan-

5-yl}-2,2-

dimethyl-1,3-

dioxolan-4-yl]-

2-(1-

ethoxyethoxy)p

ropanoate

cyano-3-

[(4S,5R)-5-

{(5S,6R)-6-[(4R)-

2,2-dimethyl-1,3-

dioxolan-4-yl]-

2,4,7,9-

tetraoxadecan-5-

yl}-2,2-dimethyl-

1,3-dioxolan-4-

yl]-2-(1-

ethoxyethoxy)pro

panoate

30

12.179

0,3815%

C27H49NOS

2

2-[(Bis{2-[(2-

methyl-2-

propanyl)sulfa

nyl]ethyl}amin

o)methyl]-4,6-

bis(2-methyl-2-

propanyl)phen

ol

2-[(Bis{2-[(2-

methyl-2-

propanyl)sulfanyl

]ethyl}amino)met

hyl]-4,6-bis(2-

methyl-2-

propanyl)phenol

-

31

12.397

1,5741%

C25H45NO9

Pederin

(2S)-N-[(S)-

{(2S,4R,6R)-6-

[(2S)-2,3-

Dimethoxypropyl

]-4-hydroxy-5,5-

dimethyltetrahydr

o-2H-pyran-2-

yl}(methoxy)met

hyl]-2-hydroxy-2-

[(2R,5R,6R)-2-

methoxy-5,6-

dimethyl-4-

methylenetetrahy

dro-2H-pyran-2-

yl] acetamide

Anticancer

(ghoneim, 2013)

32

12.614

1,9858%

C33H59NO1

4

2-(aziridin-1-

yl)ethanol;

decanedioic

acid; 2,2-

dimethylpropa

ne-1,3-diol; 2-

ethyl-2-

(hydroxymethy

l)propane-1,3-

diol;

isophthalic

acid

-

Metabolite Proﬁling of 96

27

33

12.797

2,5108%

C29H39N7O

2

1-(2-

Methylalanyl-

5-phenyl-D-

norvalyl)-4-{2-

[2-(2H-

tetrazol-5-

yl)ethyl]phenyl

}piperidine

1-(2-

Methylalanyl-5-

phenyl-D-

norvalyl)-4-{2-

[2-(2H-tetrazol-5-

yl)ethyl]phenyl}p

iperidine

-

34

13.208

0,9465%

C30H53NO1

2

(3S)-16-{[(1S)-

1-

Carboxyethyl]a

mino}-2-

methyl-16-oxo-

3-hexadecanyl

6-O-(3-

carboxypropan

oyl)-β-D-

glucopyranosid

e

(3S)-16-{[(1S)-1-

Carboxyethyl]ami

no}-2-methyl-16-

oxo-3-

hexadecanyl 6-O-

(3-

carboxypropanoyl

)-β-D-

glucopyranoside

-

35

13.460

2,6423%

C29H45N5O

2

8-

(Benzylamino)

-7-hexadecyl-

3-methyl-3,7-

dihydro-1H-

purine-2,6-

dione

8-(Benzylamino)-

7-hexadecyl-3-

methyl-3,7-

dihydro-1H-

purine-2,6-dione

-

36

13.677

2,4722%

C28H46N5O

2Cl

N4-(5-Chloro-

2,4-

dimethoxyphen

yl)-N6-

hexadecyl-

4,5,6-

pyrimidinetria

mine

N4-(5-Chloro-

2,4-

dimethoxyphenyl

)-N6-hexadecyl-

4,5,6-

pyrimidinetriamin

e

-

37

14.409

10,3549%

C25H50NO6

Cl

-

38

14.740

2,0423%

C22H48N9Cl

N2-[3-({12-

[(3-

Aminopropyl)a

mino]dodecyl}

amino)propyl]-

N4-methyl-

1,3,5-triazine-

2,4,6-triamine

hydrochloride

(1:1)

N2-[3-({12-[(3-

Aminopropyl)ami

no]dodecyl}amin

o)propyl]-N4-

methyl-1,3,5-

triazine-2,4,6-

triamine hydrochl

oride (1:1)

-

39

15.106

23,3199%

C8H39N23O

-

40

15.404

4,7166%

C24H50N9Cl

-

41

15.769

1,1138%

C8NO15S6B

-

28

r2

42

15.952

0,6060%

C8NO15S6B

r2

-

43

16.718

5,9510%

C36H36N5O

6SCl

4-[(N-{2-[(6-

Chloro-2-

methyl-4-

quinolinyl)ami

no]ethyl}-N-

[(4-

methoxyphenyl

)sulfonyl]-β-

alanyl)amino]-

3-methoxy-N-

phenylbenzami

de

4-[(N-{2-[(6-

Chloro-2-methyl-

4-

quinolinyl)amino]

ethyl}-N-[(4-

methoxyphenyl)s

ulfonyl]-β-

alanyl)amino]-3-

methoxy-N-

phenylbenzamide

-

44

17.004

1,3681%

C7H24N19O

9Cl

-

45

17.999

4,6577%

C46H48N5O

S4Cl

-

46

18.330

11,9297%

C8NO15S6B

r2

-

47

21.509

0,0036%

-

48

21.726

0,0049%

-

49

22.389

0,0047%

-

50

22.755

0,0043%

-

Table 2: Metabolite profiling Marsilea crenata Presl.in DCM blank by UPLC-QTOF-MS/MS.

No

Rt

%Area

Formula

Trivial name

IUPAC name

Activity

1

0.289

0,0032%

C11H23N4O2Cl

Tert-Butyl 4-

carbamimidamidop

iperidine-1-

carboxylate

hydrochloride

(1:1)

2-Methyl-2-

propanyl 4-

carbamimidamido-

1-

piperidinecarboxyl

ate hydrochloride

(1:1)

-

2

0.540

0,0278%

C16H22O4

Dibutyl phthalate

Antibacteri

(Khatiwora

2012),

glikosidase

inhibitor (Lee

2000),

estrogenik

(Harris 1997)

3

0.906

0,0049%

C9H22N6O2S

-

Metabolite Proﬁling of 96

29

4

1.420

0,2361%

-

5

1.786

0,0096%

C11H23NO2

11-

Aminoundecanoic

acid

11-

Aminoundecanoic

acid

-

6

1.969

0,0041%

C10H23N4O3P

Propanedioic acid,

2-[[bis(1-

methylethyl)phosp

hinyl]methyl]-,

dihydrazide

2-

[(Diisopropylphos

phoryl)methyl]mal

onohydrazide

-

7

2.084

0,0670%

C11H23NO2

11-

Aminoundecanoic

acid

11-

Aminoundecanoic

acid

-

8

2.186

0,0306%

-

9

2.632

2,8001%

-

10

4.427

0,0282%

C15H27NO5

Megalanthonine

[(1S,7R,7aR)-7-

Hydroxyhexahydro

-1H-pyrrolizin-1-

yl]methyl (2S,3S)-

2,3-dihydroxy-2-

isopropylbutanoate

antifeedant and

antifungal

(Reina 1998)

11

4.930

0,0127%

C9H21N11O

-

12

5.342

0,2477%

-

13

5.479

0,0731%

-

14

5.662

0,0912%

-

15

5.925

0,0405%

C35H41N3O

Cycloheptaneaceta

mide, N-

(phenylmethyl)-α-

[4-[(5,6,7,8-

tetrahydro-4-

methyl-9H-

pyrido[2,3-b]indol-

9-

yl)methyl]phenyl]-

N-Benzyl-2-

cycloheptyl-2-{4-

[(4-methyl-5,6,7,8-

tetrahydro-9H-

pyrido[2,3-b]indol-

9-

yl)methyl]phenyl}

acetamide

-

16

6.211

0,0164%

-

17

6.474

0,0109%

-

18

6.840

0,0031%

-

19

7.206

0,2253%

C11H16O3

1-Carboxy-3-

hydroxyadamantan

e

3-Hydroxy-1-

adamantanecarbox

ylic acid

-

20

7.457

0,0010%

-

30

21

7.640

0,0242%

C12H25NO2

Dodecanoic acid,

12-amino-

12-

Aminododecanoic

acid

-

22

8.006

0,1302%

C18H25NO

Dextromethorphan

(9α,13α,14α)-3-

Methoxy-17-

methylmorphinan

Antitussive

(Manap 1999),

anticonvulsant

(Mohseni 2016),

neuroprotective

(Zhang 2004)

23

9.504

0,0908%

C20H31NO

Trihexyphenidyl

1-Cyclohexyl-1-

phenyl-3-(1-

piperidinyl)-1-

propanol

antiparkinson

antikolinergic

(Takahashi

1999), anti

oksidan (Ji

2008)

24

9.950

0,0080%

-

25

10.967

0,5387%

26

11.448

2,3323%

C16H35N

Hexadecylamine

1-Hexadecanamine

antibacteri,

adjuvant for

diphtheria,

tetanus toxoid,

and influenza

(Attwood 2012)

27

11.630

0,3879%

C17H37NO2

2-Amino-2-

tetradecylpropane-

1,3-diol

2-Amino-2-

tetradecyl-1,3-

propanediol

-

28

11.882

0,0775%

C19H18O4

Benzylbutylphthal

ate

3-(1-Phenyl-2-

pentanyl)phthalate

Estrogenik

(Harris 1997)

29

12.111

0,0640%

C17H26O5

Portentol

(1S,2S,3S,3'R,4R,4

'R,5'S,6'R,8R)-4'-

Hydroxy-

1,3,3',5',6',8-

hexamethyltetrahy

dro-6H,7H-

spiro[5-

oxabicyclo[2.2.2]o

ctane-2,2'-pyran]-

6,7-dione

Anticancer

(Schröckeneder

2012)

30

12.248

0,0123%

C15H33N

Pentadecylamine

1-

Pentadecanamine

-

31

12.396

0,0027%

C19H41NO2

1,2-Propanediol, 3-

(hexadecylamino)-

3-

(Hexadecylamino)-

1,2-propanediol

-

32

12.694

0,6293%

C19H18O4

Benzylbutylphthal

3-(1-Phenyl-2-

Estrogenik

Metabolite Proﬁling of 96

31

ate

pentanyl)phthalate

(Harris 1997)

33

12.842

0,9778%

C21H37N

4-

Pentadecylaniline

4-

Pentadecylaniline

-

34

13.894

0,9962%

C23H41N

Benzylamine,

N,N-dioctyl-

N-Benzyl-N-octyl-

1-octanamine

-

35

15.072

16,7611

%

C12H21N25O5

S

-

36

15.323

6,5543%

C12H21N25O5

S

-

37

15.987

26,3455

%

C38H38N5O11

Cl

(1R,13S,16S,17R,2

8R)-28-Amino-20-

chloro-17,25-

dihydroxy-

5,8,10,24-

tetramethoxy-N-

methyl-15,29,31-

trioxo-22-oxa-

14,30,32-

triazahexacyclo[14

.14.2.2

18,21

.1

2,6

.1

23,2

7

.0

7,12

]hexatriacont

a-2(36),3,5

,7,9,11,18,20,23(3

3),24,26,34-

dodecaene-13-

carboxamide

(1R,13S,16S,17R,2

8R)-28-Amino-20-

chloro-17,25-

dihydroxy-

5,8,10,24-

tetramethoxy-N-

methyl-15,29,31-

trioxo-22-oxa-

14,30,32-

triazahexacyclo[14

.14.2.2

18,21

.1

2,6

.1

23,2

7

.0

7,12

]hexatriacont

a-2(36),3,5

,7,9,11,18,20,23(3

3),24,26,34-

dodecaene-13-

carboxamide

-

38

17.050

0,4132%

-

39

17.599

1,9907%

C35H36N4O5

Pheophorbide A

3-[(3S,4S,21R)-14-

Ethyl-21-

(methoxycarbonyl)

-4,8,13,18-

tetramethyl-20-

oxo-9-vinyl-3-

phorbinyl]propanoi

c acid

Anticancer

(Cho, 2014)

40

18.433

37,6384

%

C36H36N5O6S

Cl

Benzamide, 4-[[3-

[[2-[(6-chloro-2-

methyl-4-

quinolinyl)amino]e

thyl][(4-

methoxyphenyl)sul

fonyl]amino]-1-

oxopropyl]amino]-

3-methoxy-N-

phenyl-

4-[(N-{2-[(6-

Chloro-2-methyl-

4-

quinolinyl)amino]e

thyl}-N-[(4-

methoxyphenyl)sul

fonyl]-β-

alanyl)amino]-3-

methoxy-N-

phenylbenzamide

-

32

41

19.645

0,0049%

-

42

20.960

0,0047%

-

43

21.109

0,0065%

C12N

-

44

21.326

0,0063%

-

45

21.509

0,0074%

-

46

21.658

0,0150%

C12N

-

47

22.572

0,0466%

C7H10N2

2-

Pyridylethylamine

2-(2-

Pyridinyl)ethanami

ne

-

The largest compound in 96% ethanol extract

leaves Marsilea crenata Presl. on methanol blank

with % area 23,3199 %; 11.9297% and 10.3549%

are unknown compounds where the chemspider

application does not recognize it or has never been

published. Whereas in the DCM blank on % area

37.6384 % is C

36

H

36

N

5

O

6

SCl after data

interpretation was done by using website

Chemspider and software Chemdraw so that

compound similarity 4-[(N-{2-[(6-Chloro-2-methyl-

4-quinolinyl)amino] ethyl}-N-[(4-methoxyphenyl)

sulfonyl] -β-alanyl) amino] -3-methoxy-N-

phenylbenzamide was obtained ; peak area 26.3455

% is C

38

H

38

N

5

O

11

Cl and suitable with compound

(1R, 13S, 16S, 17R, 28R) -28-Amino-20-chloro-

17,25-dihydroxy-5,8,10,24-tetramethoxy-N-methyl-

15,29, 31-trioxo-22-oxa-14,30,32-triazahexacyclo

14.14.2.218,21.12,6.123,27,07,12] hexatriaconta-2

(36), 3,5, 7,9,11,18,20,23 (33), 24,26,34-dodecaene-

13-carboxamide and we did not obtaine the

compound name that was not suitable with the

compound name reference. So that we catagorized

as unknown compound.

The activity of the major compound explained

above had non been obtained yet before. According

to the research was done, it need to analyzed deeply

in order to get the data about unknown compound.

4 CONCLUSIONS

From the analysis data, we can conclude that there

are some phytochemical compound in Marsilea

crenata Presl. leaves that was known having major

unknown compound.

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