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Table 1 Tentative assignment of HPLC chromatogram peaks as flavonoids, phenylcarboxylic acids or diterpenes based on UV absorbance and MS fragmentation patterns

From: An in vitro study of anti-inflammatory activity of standardised Andrographis paniculata extracts and pure andrographolide

Peakidentitycor number

Rt (min)

% Area3

Fold variation2

UV peaks1(nm)

MS fragmentation (m/z)5

Tentative assignment4

Chlorogenic acid

12.2

1.0

34.3

327, 218, 235

Not determined (ND)

Phenyl carboxylic acid

Isoquercetin

28.7

0.9

5.1

204, 255, 353

ND

Flavonol glycoside

Peak 3

29.2

0.6

9.8

323, 219, 234

ND

Phenyl carboxylic acid

Peak 4

29.7

3.5

3.8

325,219,235

ND

Phenyl carboxylic acid

Peak 5

31

1.4

3.4

326, 219, 271

ND

Phenyl carboxylic acid

Peak 6

32.2

4.1

6.1

324, 219, 234

ND

Phenyl carboxylic acid

Peak 7

32.4

3.3

4.5

325, 220

ND

Phenyl carboxylic acid

Peak 8

32.8

3.3

4.7

346, 224, 256

ND

Phenyl carboxylic acid

Peak 9

33.7

2.1

4.5

204, 336, 266

ND

Flavone

Peak 10

36.8

1.3

6.6

327, 219, 235

ND

Phenyl carboxylic acid

Peak 11

37.7

0.8

4.4

327, 271, 223, 204

ND

ND

Andrographolide

38.4

ND

ND

227

351 [M+H]+, 333 [M-H2O]+, 315 [M-2H2O]+, 297 [M-3H2O]+

Diterpene

Andropanoside

39.8

1.4

4.8

207

535 [M+K]+

Diterpene

Peak 14

40.2

8.0

7.1

225

ND

ND

Peak 15

40.7

4.3

3.7

202

ND

ND

Apigenin

42.2

2.6

6.7

211, 337, 267

ND

Flavone

Wogonoside

45.2

0.7

2.3

265, 212

ND

O-methylated flavone glycoside

Peak18

46.6

0.7

4.6

ND

ND

ND

Peak 19

46.8

0.8

2.5

ND

ND

ND

Neoandrographolide

47.7

9.3

2.7

201

503 [M+Na]+,519 [M+K]+, 319[M+H-Glu]+

Diterpene

Peak 21

48.6

0.5

3.5

ND

ND

ND

Peak 22

49.7

1.5

5.3

ND

ND

ND

Peak 23

50.2

0.76

7.7

ND

ND

ND

Deoxyandrographolide

52.1

6.7

3.7

200

357 [M+Na]+, 317 [M+H-H2O]+, 299 [M+H-2H2O]+

Diterpene

Dehydroandrographolide

52.6

35.1

2.1

200, 249

355 [M+Na]+, 315 [M+H-H2O]+, 297 [M+H-2H2O]+

Diterpene

Peak 26

53.6

0.8

2.8

229

ND

ND

Peak 27

54.3

2.0

6.1

229

ND

ND

Peak 28

54.6

2.8

3.0

200, 263

ND

ND

  1. 1UV peaks are listed in order of intensity.
  2. 2Fold variation = (Max. peak .area) / (Min. peak area).
  3. 3Percent (%) area = average peak area from the 12 batches / total combined area × 100. Andrographolide was excluded from the % area calculations.
  4. 4Assignments based on UV spectrum, MS fragmentation and/or comparison to reference standard RT.
  5. 5MS fragments are listed in order of intensity.