LC-MS Application Data Sheet

No. 050

Analysis of aflatoxins using LCMS-2010A

fluorescence detection and LC-MS were adopted as

aflatoxin analysis methods.
This data sheet introduces an example of aflatoxin
analysis using LC-MS in accordance with the official
methods. Figs. 1 and 2 show SIM chromatograms of
four aflatoxin components G2, G1, B2 and B1
(2.5ppb and 0.5ppb respectively). Good results were
obtained for each component (S/N= 203 to 557 at
2.5ppb and S/N= 43 to 84 at 0.5ppb). Fig. 3 shows
the mass spectra and calibration curves (0.5ppb to
100ppb, n=5) for each component. Good linearity at
r2= 0.9999 or higher was obtained for each
component.

Aflatoxins are naturally occurring toxic

substances with strong carcinogenicity produced by
mold (aspergillus) growing on peanuts, corn and
other grains. Japanese Ministry of Health, Labor and
Welfare announced aflatoxin testing methods in the
1971. The Food Sanitation Law in Japan controls
food products from which aflatoxins exceeding 10ppb
is detected. This method of aflatoxin testing was
revised on March 26, 2002 into methods that do not
use harmful reagent. As a result HPLC with

Int.

Based on
Official Method

35000

313.00(1.22)
315.10(1.00)
329.10(1.00)
331.10(1.00)

G2
S/N=557

32500

30000

27500

G1
S/N=398
B2
S/N=296

25000

22500

B1
S/N=203

20000

17500

15000

12500

[M+H]+

10000

[M+H] m/z 331

m/z 329

7500

[M+H]+

[M+H]+

5000

m/z

m/z

315
2.5

5.0

7.5

10.0

12.5

313

15.0

17.5

20.0

min

Fig. 1 SIM chromatogram of aflatoxin mixture (each 2.5ppb)

Int.

Based on
Official Method

9500

313.00(1.41)
315.10(1.21)
329.10(1.08)
331.10(1.00)

G1
S/N=60

G2
S/N=8

9000

8500

B2
S/N=62

B1
S/N=43

8000

7500

7000

6500

6000

[M+H]+ m/z 315

5500

5000

4500

[M+H]+

4000

331

[M+H]+ m/z 313

[M+H]+ m/z 329

m/z

3500

3000

2.5

5.0

7.5

10.0

12.5

15.0

17.5

20.0

Fig. 2 SIM chromatogram of aflatoxin mixture (each 0.5ppb)

min

Int.
1250e3

Aflatoxin G2

Area
17.5e6

331.1

MW 330

15.0e6

1000e3

12.5e6
10.0e6

750e3

7500e3

500e3

369.0

5000e3

353.1

250e3

229.0 246.0

0e3202.2
200

225

313.3

284.6

250

275

300

389.1

362.1
325

350

375

m/z

Aflatoxin G1

329.1

r2=0.99998
0

25

50

75

Conc

15.0e6

MW 328

700e3

0e3

Area
17.5e6

Int.
800e3

2500e3

399.1

12.5e6

600e3
500e3

10.0e6

400e3

7500e3

300e3

5000e3

200e3

351.1

100e3

215.3
0e3

247.3

225

282.9

250

275

311.0
300

333.4
325

374.1
387.1
391.9

350

375

m/z

315.1

Aflatoxin B2

0e3

r2=0.99999
0

25

50

75

Conc

Area
17.5e6

Int.
1000e3

2500e3

MW 314

15.0e6
12.5e6

750e3

10.0e6
500e3

7500e3
250e3

0e3

Int.

5000e3

360.1

225

250

336.9 353.1 373.1

287.1 306.1

242.7 259.0

213.0

275

300

325

350

2500e3

r2=0.99999

388.4

375

m/z

0e3

25

50

75

Conc

Area

Aflatoxin B1

15.0e6

313.0

MW 312

12.5e6

750e3

10.0e6
500e3

7500e3
5000e3

250e3

0e3
200

358.3
213.7
225

240.9

335.1 350.9 371.1

268.7 284.7

250

275

300

325

350

2500e3

r2=0.99999

385.9

375

m/z

0e3

25

50

75

Conc

Fig. 3 Mass spectra and calibration curves (0.5-100ppb, n=5) of aflatoxins G2,G1,B2 and B1
Column
Mobile phase A
Flow rate
Injection volume
Probe voltage
CDL temperature
Nebulizing gas flow
CDL voltage
Q-array DC voltage
Scan range
SIM monitoring ions

Table 1 Analytical conditions of LC-MS

: Imtakt Cadenza CD-C18 (2.0 mmI.D. x 150 mm)
: acetonitrile / methanol /10mM ammonium acetate = 2 / 6 / 15
: 0.2 mL/min
: 6L
Column temperature
: 40 C
: 4.5kV (ESI-Positive mode)
: 300 C
BH temperature
: 250 C
: 1.5 L/min
Drying gas flow
: 0.15MPa
: 25 V
: Scan-mode
Q-array RF voltage
: Scan-mode
: m/z 200-400 (1.0sec/scan)
: m/z 331.1, 329.1, 315.1, 313.0 (0.25sec/ch)

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