Artemisia annua

Chinese name: Qing hoa su English: wormwood, sweet wormwood

Artemisia annua

Crystals of artemisinin isolated from A. annua

Aug. 19, 2005

17

2.3

97

10

5.7

80

94

.11

8

79

.91

27

7.7

43

50

.52

1

45

.42

1

37

.95

13

6.3

47

34

.04

63

3.3

15

25

.60

52

5.2

84

23

.82

12

0.2

22

12

.95

7

O

O

O

H

H

Me

OO

Me

Me

H

H

Artemisinin

105.7

94

172.3

Dept-135, and C-13 NMR Spectra of Artemisinin

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

12.956

20.224

23.809

25.265

25.599

30.089

33.302

34.022

36.320

37.936

45.394

50.490

77.095

77.412

77.617

77.730

79.904

94.112

105.785

172.448

C10

C12

C3

C12

aX

O

O

O

H

H

Me

OO

Me

Me

H

H

Artemisinin

103 12

12a

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

29.543

29.642

29.733

29.818

29.997

30.073

30.311

32.299

32.582

33.066

33.295

33.998

35.639

36.291

36.743

37.901

40.068

42.119

42.510

43.181

43.454

43.685

44.003

45.352

50.455

58.760

61.228

63.398

77.125

77.443

77.645

77.760

79.886

94.141

105.782

119.730

122.108

130.588

136.315

172.624

182.902

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-12C-12a

C-10

C-13 NMR spectrum of the crude EtOH ext of A. annua

O

O

O

H

H

Me

OO

Me

Me

H

H

3

5 6

8a

8

9

10

12

12a

15a

C-3

Aug. 19, 2005

Aug. 19, 2005

1 2 3

1. EtOH Ext2. Artemisinin3. Tea (5 g)

Dihydro-epideoxyareannuin B

Artemisinin

Confirms presence of Active ingredient in the Tea

17

9.6

83

14

2.5

84

12

2.2

04

83

.55

7

47

.00

44

3.2

28

40

.04

1

32

.84

03

1.2

71

30

.09

73

0.0

56

24

.14

92

3.8

20

21

.43

02

0.0

11

9.8

32

(S)

(S)

(S)

(R)

O

(R)

O

H

(Z)

2

5

7

10

14

15

1

1112

13

Dihydro-epideoxyarteannuin B

Constituents other than Artemisinin

•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

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)

Source: Planta Medica, 1993 (I.d. 16624)

Quantitative Aspects

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.

O

O

O

H

H

Me

OO

Me

Me

H

H

1H NMR of Artemisinin

7.26

1

5.85

8

3.72

73.

709

1.44

21.

241

1.00

70.

993

1.00

2.31

0.92

0.95

3.86

6.09

3

5.37

55.

373

1.00

1.03

2.88

0.98

1.01

1.011.001.01

1.08

ppm (f1)5.005.506.00

6.09

3

5.37

55.

373

1.00

1.03

1.01

1.01

1.00

1.01

H

HO

O

7.26

1

6.20

16.

192

5.83

3

Ref. DHCL

Top: 8 month plant extract 77-75I

Bottom: Artem.

1.00

2.35

Expansion of H-12 region of 77-75I

Ref. 2 mg0.0087 mmol

From ratioArtem. mmol = 0.019Hence Artem. Wt = 5.47 mgArtem % = 0.6%

H-12

ppm (f1)6.00

1.00

1.05

2 month plant extract 77-76C

Integral Ratio 1:1Calcd Artem. mmol = .009Hence Artem. Wt = 2.5 mgArtem % = 0.3%

Ref. 2 mg0.0087 mmol

ppm (f1)5.506.006.507.007.50

1.00

0.80

0.37

Artemisinin 1 mg 282 MW 0.0035 mmolCaffeine 2mg 194 MW 0.0100 mmolDHCL 2mg 230 MW 0.0087 mmol

Artemi/Caf. Integral Ratio .037/1Therefore Artemi = 0.37 x 0.01 = 0.0037 mmolMass 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 mmolMass of Artemi. = 0.004 x 282 = 1.1 mg = 1 mg

Conclusion: Actual Artemi. Added 1 mg Found by QHNMR 1 mg

What next?

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