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Uv Vis Analysis
Uv Vis Analysis
249
250
251
TABLE 1
Comparison of Mean Duplicate Samplesa
acetone-SP825
hexane-SK104
isopropanol-SK104
modes of the C-H groups. The weak absorption band observed at 3005 cm1 is due to the trans-CH=CH- of the
-carotene. The sharp band at 1455 cm1 arose from the
asymmetric deformation mode, whereas the band at 1373
cm1 was the symmetric deformation mode of the C-H group.
At lower frequencies, the band at 956 cm1 corresponded to
the trans conjugated alkene -CH=CH- out-of-plane deformation mode (17), whereas the weak band near 727 cm1 was
assigned to the CH2 group in rocking mode.
Figure 3 represents overlaid spectra of CPO, containing
~200800 ppm -carotene, centered on the trans-CH=CH- of
the -carotene. A PLS calibration was constructed and optimized based on the FTIR spectral region from 976 to 926 cm
1
referenced to a single-point baseline at 926 cm1. The selection and optimization of the wavelength range, type of
baseline configuration, and number of factors were guided by
the PRESS test. Figure 4 illustrates the calibration plot obtained from the PLS calibration model using seven factors in
terms of FTIR predicted vs. actual spectrophotometric carotene for 30 samples (Table 1). The plot shows overall linear regression with R2 of 0.972 and SE of 25.8 ppm. When
the calibration was examined with the leave-one-out cross
validation, the overall SE was 29.6 ppm.
NIR spectroscopy. Figure 5 represents a mean spectrum of
CPO (30 samples) in the NIR region, which was more difficult to interpret mainly because of the extensive band overlapping due to the overtone and combination of the funda-
FTIR method
NIR method
528
530
535
535
526
527
526
537
503
542
503
539
553
532
794
733
692
574
343
360
301
222
261
304
257
248
272
266
531
520
559
546
562
565
563
395
393
346
388
388
282
299
287
239
281
528
527
546
528
530
528
543
533
519
549
519
500
554
528
751
NA
651
564
365
411
334
294
300
333
245
NA
288
243
528
NA
567
568
580
587
576
379
379
361
379
362
249
278
NA
NA
249
535
542
537
532
513
530
477
511
500
479
500
518
535
517
782
755
699
547
355
380
260
321
301
347
306
301
285
268
546
543
537
533
586
567
556
316
426
315
353
379
311
319
310
310
318
a
UV, ultraviolet; FTIR, Fourier transform infrared spectroscopy; NIR, near
infrared spectroscopy; NA, not available.
252
1.0
1455
956
0.9
1373
0.8
0.7
3005
0.6
0.5
0.4
727
0.3
0.2
0.1
4000
3500
3000
2500
2000
1500
1000
500
the trans-CH=CH- functional group. However, this characteristic band did not exist in the NIR region (20). The correlation spectrum (Fig. 6) also revealed that there was a mathematical correlation between spectral changes with the
changes in the -carotene content. However, there was strong
correlation between 500 and 700 nm in the visible region.
FIG. 4. Plot of mean carotene content obtained from calibration standards by ultraviolet (UV)-visible spectrophotometry reference method
vs. FTIR method. See Figure 2 for abbreviation.
1.14
1.12
1.10
1.08
1.06
1.04
1.02
1.00
Absorbance
0.98
0.96
0.94
0.92
0.90
0.88
2.6
2.4
0.86
0.84
2.2
2.0
0.82
1.8
1.6
0.80
1.4
1.2
0.78
1.0
0.8
0.76
980
970
960
950
940
0.6
0.4
0.2
Wavenumbers (cm )
FIG. 3. Overlaid spectra of crude palm oil samples showing different
concentrations of carotene using FTIR spectroscopy. See Figure 2 for
abbreviation.
400
600
800
1000
1200
1400
1600
1800
2000
2200
2400
253
0.60
0.55
0.50
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0.00
400
600
800
1000
1200
1400
1600
1800
2000
2200
2400
from the validation sets are 25.78 for FTIR method and 19.96
for NIR method. These plots are linear, with R2 of 0.951 for
FTIR technique and 0.979 for NIR technique, respectively. The
regression lines were:
FTIR -carotene = 30.401 + 0.9144 UV-vis -carotene
[1]
[2]
FIG. 7. Plot of mean carotene content obtained from calibration standards by UV-visible spectrophotometry reference method vs. NIR
method. For abbreviations see Figures 4 and 5.
TABLE 2
Statistical Comparisona of Carotene Data Obtained by UV-Visible
Spectrophotometer with FTIR and NIR Methods
Statistic
Max. value
Min. value
Mean
MDr
SDDr
MDa
SDDa
UV-visible
reference method
795
223
452
2.67
1.67
3.78
0.69
FTIR
method
NIR
method
751
243
456
6.45
2.37
6.44
7.27
783
260
448
9.11
8.95
FIG. 8. Plot of mean FTIR carotene content vs. mean reference carotene
content of validation samples. For abbreviation see Figure 2.
254
5.
6.
7.
8.
9.
10.
11.
12.
FIG. 9. Plot of mean NIR carotene content vs. mean reference carotene
content of validation samples. For abbreviation see Figure 5.
ACKNOWLEDGMENTS
The authors thank the Ministry of Science, Technology and Environment of Malaysia for providing IRPA funds (0302040048). Gratitude is also expressed to Prof. F.R. van de Voort of the Department
of Food Science and Agricultural Chemistry, Macdonald Campus of
McGill University, Qubec, Canada, for his technical assistance.
13.
14.
15.
16.
REFERENCES
1. Jacobsberg, B., Palm Oil Characteristics and Quality, in Proceedings of 1st Malaysian Agricultural Research and Development Institute Workshop on Oil Palm Technology, Kuala
Lumpur, Malaysia, pp. 4868 (1974).
2. La Vecchia, C., A. Decarli, S. Franceshi, A. Gentile, E. Negri,
and F. Parazzini, Dietary Factors and the Risk of Breast Cancer,
Nutr. Cancer 10:205214 (1987).
3. Seifter, E., G. Rettura, J. Padawer, and S.M. Levenson, Moloney
Marine Sarcoma Virus Tumors in CBA/J Mice: Chemopreventive and Chemotherapeutic Actions of Supplemental Carotene, J. Natl. Cancer Inst. 68:835840 (1982).
4. Alam, B.S., and S.Q. Alam, The Effect of Different Levels of
17.
18.
19.
20.