Rating of units 2 and 3

Metabolites derived from the pathways of chikimic acid

and acetyl coenzyme A
Exam Units 2 and 3: Natural Products.

Diego Federico Rodríguez Vélez CC 1000556666

list of participants: Individual assignment of model compound

Item Email Exam Units 2 and 3

1 muneton.catalina@uces.edu.co β-sitosterol
2 munozgu.sara@uces.edu.co naringenin
3 laura.valenciat@uces.edu.co Curcumin
4 gaviria.paola@uces.edu.co campestanol
5 raffaeli.juan@uces.edu.co Demethoxycurcumin
6 palacios.yessica@uces.edu.co Hesperetin
7 sofia.torresv@uces.edu.co cycloartenol
8 gomezc.valentina@uces.edu.co Apigenin
9 solling.melani@uces.edu.co Diosgenin
10 echeverria.daniel@uces.edu.co Sesamin
11 pamplona.juan@uces.edu.co campesterol
12 vasco.valentina@uces.edu.co Bisdemethoxycurcumin
13 alfonso.danna@uces.edu.co Hecogenin
14 gutierrez.yamile@uces.edu.co Eriodictyol
15 valencia.yiset@uces.edu.co stigmasterol
16 celis.maria@uces.edu.co medioresinol
17 atehortua.dayana@uces.edu.co lupeol
18 jaramillo.annie@uces.edu.co Sakuranetin
19 rojas.elizabeth@uces.edu.co Syringaresinol
20 rodriguez.diego@uces.edu.co stigmastanol
21 vellojin.maria@uces.edu.co Nobiletin
22 garcia.juanc@uces.edu.co Artemisinin
23 cruz.laura@uces.edu.co β-carotene
24 monsalve.geraldine@uces.edu.co Cynaropicrin
25 rojas.andres@uces.edu.co ergosterol
26 borda.laura@uces.edu.co Helenalin
27 montoyata.maria@uces.edu.co β-cryptoxanthin
28 gomez.laura@uces.edu.co brassicasterol
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
Value of this evaluation (5 points)

According to the activity "Units 1 and 2 Paper review" and the substance assigned in the table; you must
propose the following:
1. (1.0) Report at least five plant species that contain the assigned metabolite. propose a method for the
extraction of this substance from a species mentioned above. (Reference with link)

a. Genetically modified Soy bean plants (Glycine max.) From Dewick P. Medicinal Natural Products: A
Biosynthetic Approach: Third Edition. 3rd ed. Chichester: John Wiley & Sons Ltd; 2009. Page 255.
I would recommend to first let the newly introduced oxydase make effecton present stigmasterols
and then extract using some mineral oil or glycols.
*Most of this extractions would take place on a HPLC system preferably .

b. False yellowhead (Dittrichia viscosa) From

https://journals.sagepub.com/doi/pdf/10.1177/1934578X0800301110
I would recommend to only macerate the leaves and then extract using acetone-water or mineral
oils.

c. Bilimbi tree (Averrhoa bilimbí) From

https://journals.sagepub.com/doi/pdf/10.1177/1934578X1501000116
I would macerate the leaves and then extract using hexane-etanol.

d. Kava Kava (Piper Methysticum) From

https://journals.sagepub.com/doi/pdf/10.1177/10915818093509341
It´s proposed to macerate the roots and then extract metabolites using etanol-water or acetone
water.

e. Marigold (Calendula officinalis) From

https://journals.sagepub.com/doi/pdf/10.1177/1091581810384883
They purposed a variety of glycols and mineral oils to extract lipophilic substances out of marigold´s
stem and flower, including stigmastanol with a presence of 0.2%

2. (0.5) Express the name of the compound according to IUPAC, in addition to its formula and exact mass.
(Reference with link).

 IUPAC name: (3S,5S,8R,9S,10S,13R,14S,17R)-17-[(2R,5R)-5-ethyl-6-methylheptan-2-yl]-10,13-

dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol
 Molecular formula: C 29 H 52 O
 Molecular weight: 416.7 g/mol
From https://pubchem.ncbi.nlm.nih.gov/compound/Stigmastanol

3. (0.5) Consult the structure (stereo model) with absolute configuration and add a 3D image of the most
stable conformer of the assigned metabolite. (Reference with link)
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A

Stereo structure of stigmastanol

3D structure of stigmastanol

Info from https://pubchem.ncbi.nlm.nih.gov/compound/Stigmastanol

4. (1.5) Using the simulator: https://www.nmrdb.org/

Draw the structure correctly, simulate the H-NMR spectrum (600 Mhz; 0-14; 64 k; line width: 1 Hz), take
an image (or several images, if you consider it important to enlarge any area of the spectrum) of the
spectrum in full screen and add this to your answer. A table must be prepared with the data for chemical
displacement, coupling constants, multiplicity and integral for each of the signals. In this table a column
must be placed to indicate observations, explanation and/or comments on the behavior of the
respective signals. (Reference with link)
for example:
R/.
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A

*How I´m counting stigmastanol´s carbons.

In general terms, proton signals between 1 and 2 ppm are from the hydrogens of the steroidal skeleton
of the molecule and other secondary and tertiary carbons present in the posterior hydrocarbon tail of
the compound, implying that this area is full of weak and not well defined signals, as showed in the
graph below the table. The area above and close to 2 ppm belongs to protons nearby the hydroxyl
group, like the ones from carbons 2, 3 and 4 of the steroidal skeleton, and have a weak signal, all that
because electronegative groups like hydroxyl increase the chemical shift and weakens magnetic
resonance. The area below 1 ppm belongs to the hydrogens from the methyl groups, with a well-defined
strong signal, also hydrogens from secondary carbons 22 and 23 in the hydrocarbon tail have their
signals there, being weaker and the having the lowest shift.

Position  Simulation 1H-NMR 600 MHz Observations

δh (ppm) Nh Multiplicity J(Hz)

A hydrogen that belongs to
30 1.3 1 tt 10.26,2.78 the 5th carbon, its positioned
backwards.
A hydrogen that belongs to
31 1.664 1 ddd 8.03,4.63,4.31 the 17th carbon, its
positioned backwards.
A hydrogen that belongs to
32 1.365 1 qd 10.22,2.8 the 8th carbon, its positioned
forwards.
A hydrogen that belongs to
33 1.232 1 ddd 10.25,10.2,3.22 the 9th carbon, its positioned
forwards.
A hydrogen that belongs to
34 1.602 1 ddd 10.22,9.71,4.88 the 14th carbon, its
positioned forwards.
A hydrogen that belongs to
35 1.792 1 dt 13.036,2.79
the 1st carbon.
A hydrogen that belongs to
36 1.703 1 ddd 13.036,10.26,2.79
the 1st carbon.
A hydrogen that belongs to
37 2.11 1 dt 13.357,2.78
the 4th carbon.
A hydrogen that belongs to
38 1.619 1 dt 13.357,10.26
the 4th carbon.
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
A hydrogen that belongs to
39 1.563 1 dtd 13.218,10.26,2.79
the 2nd carbon.
A hydrogen that belongs to
40 1.8 1 dq 13.218,2.78
the 2nd carbon.
A hydrogen that belongs to
the 3rd carbon. Heavily
41 3.592 1 tt 10.26,2.78
shifted by neighboring
hydroxyl group.
A hydrogen that belongs to
42 1.211 1 dtd 13.1,10.26,2.78
the 6th carbon.
A hydrogen that belongs to
43 1.688 1 dq 13.1,2.78
the 6th carbon.
A hydrogen that belongs to
44 1.277 1 dtd 13.016,10.26,2.78
the 7th carbon.
A hydrogen that belongs to
45 1.79 1 dq 13.016,2.78
the 7th carbon.
A hydrogen that belongs to
46 1.586 1 dddd 13.076,3.22,2.96,2.53
the 11th carbon.
A hydrogen that belongs to
47 1.499 1 dtd 13.076,10.2,3.17
the 11th carbon.
A hydrogen that belongs to
48 1.455 1 ddd 13.095,10.24,2.96
the 12th carbon.
A hydrogen that belongs to
49 1.588 1 ddd 13.095,3.17,2.53
the 12th carbon.
A hydrogen that belongs to
50 1.562 1 dddd 13.915,9.71,7.7,6.79
the 15th carbon.
A hydrogen that belongs to
51 1.533 1 dddd 13.915,6.95,4.88,1.73
the 15th carbon.
A hydrogen that belongs to
52 1.654 1 dddd 13.752,6.79,4.63,1.73
the 16th carbon.
A hydrogen that belongs to
53 1.676 1 dddd 13.752,8.03,7.7,6.95
the 16th carbon.
A hydrogen that belongs to
55,56,57 0.759 3 d 6.65 the 26th carbon. A methyl
group in the tail.
A hydrogen that belongs to
the 19th carbon. A methyl
58,59,60 0.878 3 s
group positioned forwards
from the steroid.
A hydrogen that belongs to
the 18th carbon. A methyl
61,62,63 0.8770000000000001 3 s
group positioned forwards
from the steroid.
A hydrogen that belongs to
64 1.457 1 tqd 7.275,6.803,4.31
the 20th carbon in the tail.
65 0.685 1 td 7.711,7.275 A hydrogen that belongs to
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
the 22th carbon in the tail.
A hydrogen that belongs to
66 0.685 1 td 7.711,7.275
the 22th carbon in the tail.
A hydrogen that belongs to
the 21th carbon. A methyl
67,68,69 0.947 3 d 6.803
group positioned backwards
from the tail.
A hydrogen that belongs to
70 0.67 1 td 7.711,7.144
the 23th carbon in the tail.
A hydrogen that belongs to
71 0.67 1 td 7.711,7.144
the 23th carbon in the tail.
A hydrogen that belongs to
72 1.195 1 ttd 7.144,6.65,6.565
the 24th carbon in the tail.
A hydrogen that belongs to
73 1.366 1 septd 6.65,6.565
the 25th carbon in the tail.
A hydrogen that belongs to
74 1.213 1 quint 6.605
the 28th carbon in the tail.
A hydrogen that belongs to
75 1.213 1 quint 6.605
the 28th carbon in the tail.
A hydrogen that belongs to
76,77,78 0.818 3 t 6.605 the 29th carbon. A methyl
group in the tail.
A hydrogen that belongs to
79,80,81 0.759 3 d 6.65 the 27th carbon. A methyl
group in the tail.
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A

5. (1.5) Propose a complete biosynthetic pathway for the assigned metabolite, considering the explanation
of the reactions involved in the whole process. keep in mind a good bibliographic reference for this.
(Reference with link)
From https://www.kegg.jp/kegg-bin/highlight_pathway?map=map01010 and Dewick P. Medicinal Natural
Products: A Biosynthetic Approach: Third Edition. 3rd ed. Chichester: John Wiley & Sons Ltd; 2009. Page 210,
235, 236, 252
We are going to follow both Mevalonate and MEP pathways to synthesize monoterpenes, then from that we are
going to get squalene, cholesterol and finally stigmastanol, as it´s being explained by the mechanisms below.
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A
Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A

That give us stigmastanol.

Rating of units 2 and 3
Metabolites derived from the pathways of chikimic acid
and acetyl coenzyme A

6. (bonus: 1 point) Consult the literature for the spectral data of the assigned metabolite and make a
comparison with the data obtained in the simulation. Are there differences between the theoretical and
the experimental? write your explanation in the “observations” column. (Reference with link)
for example:

I could not find any information from literature about this compounds NMR spectra.

Simulation Experimental
Position  1
H-NMR 600 MHz Ref. Link:
Observations
Multiplicity, Multiplicity,
δh (ppm) δh (ppm)
J(Hz), Nh J(Hz), Nh
1
2
3
Add rows
Nh: Number of hydrogens