Page 31 - Application Handbook - Liquid Chromatography

LAAN-A-LC-E253

Application High Performance Liquid Chromatography

News Rapid Analysis of 2,4-DNPH-Derivatized Aldehydes

and Ketones Using the Prominence-i with a Shim-
No.L476 pack XR-ODS Column

Application News No. L268 presented a rapid analysis the new Prominence-i integrated high-performance
of 2,4-DNPH (2,4-dinitrophenylhydrazine)-derivatized liquid chromatograph with a 2.2 µm particle size Shim-
aldehydes and ketones using the LC-2010 integrated pack XR-ODS column. Here we present the results of
high-performance liquid chromatograph with a Shim- simultaneous analysis of thirteen 2,4-DNPH-derivatized
pack FC-ODS column. As the goal of this study was to aldehyde and ketone standards run on a conventional
obtain a shorter the analysis time than that using the 5 µm particle column, and an example of rapid analysis
previous method, we investigated the run conditions with a 2.2 µm particle column.
for 2,4-DNPH-derivatized aldehydes and ketones using

n Simultaneous Analysis of 2,4-DNPH-Derivatized
Aldehydes and Ketones mV (×10)
Fig. 1 shows the chromatogram obtained from n Peaks
1. Formaldehyde
simultaneous analysis of thirteen 2,4-DNPH-derivatized 3.0 1 2. Acetaldehyde
3. Acetone
aldehydes and ketones using the analytical conditions 4. Acrolein

shown in Table 1. With the conventional VP-ODS 2.5 5. Propionaldehyde
6. Crotonaldehyde

column, separation of the thirteen components is 7. 2-Butanone
8. Methacrolein
complete within a 30 minute run time. The detection 2.0 9. n-Butyraldehyde

10. Benzaldehyde
limit and quantitation limit for formaldehyde were 1.5 2 4 11. Valeraldehyde
determined to be 13 pg and 41 pg, respectively. 3 12. m-Tolualdehyde
13. Hexanaldehyde
1.0 5 6 8
7 9
Table 1 Analytical Conditions 0.5 2 10 11 12 13
Column : Shim-pack VP-ODS (150 mm L. × 4.6 mm I.D., 5 µm) 7 11 13
Mobile Phase : A: Water/THF = 8/2 0.0
B: Acetonitrile 0.0 5.0 10.0 15.0 20.0 25.0 min
Time Program : B Conc. 20 % → 60 % (30 min) → 20 % (30-35 min)
Flowrate : 1.5 mL/min
Injection Volume : 10 µL
Column Temp. : 40 °C Fig. 1 Chromatogram of Standard Sample
Detection : UV 360 nm (each at 1.0 mg/L as carbonyl compounds)
n Linearity
Calibration curves were generated for standard
solutions of thirteen 2,4-DNPH-derivatized aldehydes ×10 5
and ketones (each at 0.03 – 3 mg/L, 10 µL injected)
using the Shim-pack VP-ODS. Fig. 2 shows the 7 Formaldehyde
calibration curves for formaldehyde, 2-butanone, and 2‐Butanone
2
hexanaldehyde. All three compounds produced a linear 6 Hexanaldehyde R =0.99997
plot, with an R value of greater than 0.9999 for each.
2
Table 2 shows the area reproducibility values (n=3), 5
using a concentration of 1.0 mg/L.
The area %RSD values were within 1 % for all 4 R =0.99995
2
components, indicating very good repeatability. Peak Area 3
Table 2 Repeatability of Peak Area (n=3)
2 2
Area %RSD Area %RSD R =0.99995
Formaldehyde 0.429 Methacrolein 0.186
1
Acetaldehyde 0.209 n-Butyraldehyde 0.095
Acetone 0.728 Benzaldehyde 0.979 0
Acrolein 0.108 Valeraldehyde 0.580 0 1 2 3
Propionaldehyde 0.222 m-Tolualdehyde 0.355 Concentration (mg/L)
Crotonaldehyde 0.765 Hexanaldehyde 0.644
2-Butanone 0.969 Fig. 2 Linearity