Production Practices and Quality Assessment of Food Crops. Vol. 1

Saffron Quality 227
Table 3. HPLC analysis of safranal and other volatile components of saffron.
Extraction procedure Column Elution conditions/ Reference
detection system
50% aqueous EtOH Lichrosorb Linear gradient elution Himeno and
RP-18. 5 µ (20–90% ACN in 20 min), Sano, 1987
1 ml/min, 308 nm
Saffron powder (100 mg) Bondapak C18 Gradient elution (55–90% Solinas and
+ 1 ml of int. standard ACN in 18 min, 90–99% Cichelli,
(75 ppm undecanal/ACN), ACN in 15 min), 1988
60 °C/30 min, derivatisation 0.7 ml/min, 365 nm
with 2.4-dinitrophenyl-hydrazine,
(125 mg in 50 ml ACN)
80% cold, aqueous EtOH (5 ml) Sep-PAK-C18, Linear gradient elution Sujata et al.,
centrifuge: 5000 g/10 min, 15 cm × (20–80% ACN) 1992
wash: twice with 5 ml of 4.9 mm or
80% EtOH each Isocratic elution 76% ACN
0.5 ml/min, 308 nm
Saffron (20 g) successive LiChroCart Linear gradient (20–100% Tarantilis et
extraction in Soxhlet apparatus 125 mm × ACN in 20 min), al., 1994b
with light petroleum, diethyl 4 mm 0.5 ml/min, 308 nm
ether (0.03% BHT), MeOH: Superspher
100 RP-18,
4 µ
Ground saffron (400 mg) + 50% Nova-PACK Linear gradient elution Lozano et al.,
aqueous MeOH (200 ml), stirr. RP-18, 15 cm 20–70% MeOH, 1999
for 1 h: 25 °C/dark, centrifuge: × 3.9 mm, 1 ml/min, 310 nm
30000 g/20 min 4 µ, 6 nm
100 ml of extract + 100 ml of
int. standard (0.2 mg/ml of
4-nitroaniline)
a) Saffron powder (200 mg) in Lichro-CART Isocratic elution: 20% Lozano et al.,
the extractor cell (0.5 ml) + RP-18, MeOH in 10 min 2000
0.70 ml CO 2/min 125 mm × Gradient elution: 20–70%
(17 MPa, 70 °C, 30 min) 4 mm, 5 µ, MeOH, 1%/min, 1 ml/min,
0.8 ml of extract + 0.2 ml int. 10 nm 310 nm
standard (0.2 mg/ml of
4-nitroaniline in 50% MeOH)
b) Saffron residue (200 mg) +
50% aqueous MeOH (50 ml),
stirr: 1 h/room temp./dark,
centrifuge: 30000 g/20 min
8 ml of extract + 20 ml int.
stand. (same as above)
a) Ground stigmas (100 mg) + RP-C18 Isocratic elution: 100% Loskutov et
80% aqueous EtOH (3 ml), 25 cm × ACN, 1 ml/min al., 2000
(3reps), centrifuge: 8000 rpm/ 4.9 mm, 5 µ 308 nm
5 min, wash: 80% aqueous EtOH
b) 100 mg of ground stigmas +
100% aqueous ACN (3 ml), (3 reps)
centrifuge: 8000 rpm/5 min
c) the same as in (b)
except for time of injection
(1 h after extraction)