Page 67 - Application Handbook - Liquid Chromatography

Application No.L491
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n Quantitation of β-Carotene in Red Chili Pepper separation obtained with the Nexera-e.
by LC/MS/MS Fig. 3 shows the two-dimensional separation data of
The analytical conditions are shown in Table 1, and the β-carotene obtained from DUIS-positive mode MRM
sample pretreatment conditions are shown in Fig. 2. analysis of the calibration curve, and Fig. 4 shows the
β-carotene, which is a precursor of vitamin A, was linearity of the three values (blobs) in the range of 0.01
detected in the two-dimensional separation of the to 10 mg/L, which correspond to the peak volumes
carotenoids. Quantitation was then conducted using used for quantitation. The correlation coefficient (r) =
the LCMS-8030 triple quadrupole mass spectrometer. 0.998976 indicates results with good linearity. The
Both high sensitivity and high selectivity can be quantitative result for β-carotene present in red chili
obtained using MRM analysis, and further, reduced ion pepper was calculated as 1.22 mg/L based on the
suppression can be expected with the two-dimensional concentration in the final sample following extraction.

Table 1 Analytical Conditions
1D Colum : Ascentis Cyano (250 mm L. × 1.0 mm I.D., 5 µm)
Mobile Phase : A; Hexane
B; Hexane/Butylacetate/Acetone = 80/15/5 (v/v/v)
Flowrate : 0.02 mL/min
Time Program : B Conc. 0 % (0.01 min) → 0 % (5 min) → 100 % (65 min) → 100 % (75 min) → 0 % (76 min)
Column Temp. : 30 °C
Injection vol. : 2 µL
Loop vol. : 20 µL
2D Column : Ascentis Express C18 (30 mm L. × 4.6 mm I.D., 2.7 µm)
Mobile Phase : A; acetnitrile
B; 2-propanol
Flowrate : 4 mL/min (0.8 mL/min split for MS)
Time Program : B Conc. 0 % (0.01 min) → 50 % (0.17 - 0.54 min) → 80 % (0.54 - 0.93 min) → 30 % (0.94 min) → STOP (1 min)
Detector : SPD-M30A Photo diode array detector (standard cell, wave length = 450 nm)
536.40 > 444.30)
Shimadzu LCMS-8030 (DUIS positive mode, targeted β-carotene MRM transition: m/z 536.40 > 444.30)
m/z
m/z

)PNPHFOBUFE SFE DIJMMJ QFQQFS
Capsicum annuum L. H

"EE DPOTFDVUJWF N- BMJRVPUT PG NFUIBOPM
FUIZM BDFUBUF QFUSPMFVN FUIFS W W W
Area = 551615
$PNCJOF FYUSBDUT BOE pMUSBUF Concentration = 1.22 mg/L

&WBQPSBUF UP ESZOFTT VOEFS WBDVVN $

Fig. 3 2D Plot of β-Carotene
%JTTPMWFE JO N- NFUIBOPM
UFSU CVUZM NFUIZM FUIFS W W

'JMUSBUF BOE JOKFDU µ- JOUP )1-$

Correlation
Fig. 2 Sample Preparation Coefficient (r) = 0.998976

Conc. (mg/L)

Fig. 4 Linearity of Calibration Curve for β-Carotene

Data provided by University of Messina Prof. Luigi Mondello and Chromaleont S.r.l.

First Edition: May. 2015

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