Artemisia annua

1
Artemisia annua Chinese name Qing hoa su
English wormwood, sweet wormwood
2
Artemisia annua
3
(No Transcript)
4
Crystals of artemisinin isolated from A. annua
Aug. 19, 2005
5
Artemisinin
94
105.7
172.3
6
Dept-135, and C-13 NMR Spectra of Artemisinin
12a
3
12
10
C12
C3
C12a
Artemisinin
C10
X
7

• 172.6 ppm C-10
• 105.9 and 79.9 C-3 and C-12a
• 94.1 (C-12), 50.5 (C-5a)
• and 45.3 (C-8a)

C-3
C-12a
C-12
C-10
C-13 NMR spectrum of the crude EtOH ext of A.
annua
8
Aug. 19, 2005
9
Aug. 19, 2005
Dihydro-epideoxyareannuin B

Confirms presence of Active ingredient in the Tea
Artemisinin

1. EtOH Ext
2. Artemisinin
3. Tea (5 g)

1 2 3
10
Dihydro-epideoxyarteannuin B
Constituents other than Artemisinin
11
Quantitative Aspects

• Artemisinin Levels in A. annua grown in Vietnam
• Stages of the growth of the plant
• Vegetative growth 5-8 months (from sown seeds)
• Mass formation of buds (9 months)
• Full blooming (10 months)

• Artemisinin highest in 5-months old plant (0.86)
• Content then drops to 0.4 and
• Post flowering as low as 0.16
• Essential oil 1.9 (v/w) just before flowering

Source Planta Medica, 1993 (I.d. 16624)
12
J. Nat. Prod., 68 (1), 133 -149, 2005.
Quantitative 1H NMR Development and Potential
of a Method for Natural Products Analysis
Guido F. Pauli,Birgit U. Jaki, and David C.
Lankin. College of Pharmacy, University of
Illinois at Chicago, Chicago, Illinois 60612
Abstract Based on a brief revision of what
constitutes state-of-the-art "quantitative
experimental conditions" for 1H quantitative NMR
(qHNMR), this comprehensive review contains
almost 200 references and covers the literature
since 1982 with emphasis on natural products. It
provides an overview of the background and
applications of qHNMR in natural products
research, new methods such as decoupling and
hyphenation, and analytical potential and
limitations, and compiles information on
reference materials used for and studied by
qHNMR. The dual status of natural products, being
single chemical entities and valuable
biologically active agents that need to be
purified from complex matrixes, results in an
increased analytical demand when testing their
deviation from the singleton composition ideal.
The outcome and versatility of reported
applications lead to the conclusion that qHNMR is
currently the principal analytical method to meet
this demand. Considering both 1D and 2D 1H NMR
experiments, qHNMR has proved to be highly
suitable for the simultaneous selective
recognition and quantitative determination of
metabolites in complex biological matrixes. This
is manifested by the prior publication of over 80
reports on applications involving the
quantitation of single natural products in plant
extracts, dietary materials, and materials
representing different metabolic stages of
(micro)organisms. In summary, qHNMR has great
potential as an analytical tool in both the
discovery of new bioactive natural products and
the field of metabolome analysis.
13
(No Transcript)
14
1H NMR of Artemisinin
15
(No Transcript)
16
Top 8 month plant extract 77-75I Bottom Artem.
Ref. DHCL
17
Expansion of H-12 region of 77-75I
From ratio Artem. mmol 0.019 Hence Artem. Wt
5.47 mg Artem 0.6
Ref. 2 mg 0.0087 mmol
H-12
18
2 month plant extract 77-76C
Ref. 2 mg 0.0087 mmol
Integral Ratio 11 Calcd Artem. mmol .009 Hence
Artem. Wt 2.5 mg Artem 0.3
19
(No Transcript)
20
Artemisinin 1 mg 282 MW 0.0035 mmol Caffeine
2mg 194 MW 0.0100 mmol DHCL 2mg 230 MW 0.0087
mmol


Artemi/Caf. Integral Ratio .037/1 Therefore
Artemi 0.37 x 0.01 0.0037 mmol Mass of
Artemi. 0.0037 x 282 1.04 mg 1
mg Artemi/DHCL Integral Ratio 0.37/0.8 0.46
Therefore Artemi 0.46 x 0.0087 0.004
mmol Mass of Artemi. 0.004 x 282 1.1 mg 1
mg
Conclusion Actual Artemi. Added 1 mg Found by
QHNMR 1 mg
21
What next?
22
Thank you very much for your attention