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Received 8 February 2001; received in revised form 12 July 2001; accepted 16 July 2001
Abstract
The relation between antioxidant activity and anthocyanin was determined in Roselle (Hibiscus sabdariffa L.) petals. Petals from
Roselle, cultivar F141, were collected and dried in Taitung, Taiwan. Roselle extract was prepared by extracting dried Roselle petals
in boiling water. The relation between the anthocyanin color and antioxidant capacity was elucidated by comparing absorbance at
520 nm, with ferric reducing ability of plasma (FRAP), oxygen radical absorbance capacity (ORAC) and total antioxidant status
(TAS) antioxidant assays. The results showed that the antioxidant capacity of Roselle extract increased when extraction time or
weight of petals increased. The FRAP assay showed a linear relationship with anthocyanin as determined at 520 nm. Comparisons
between FRAP and ORAC or FRAP and TAS assays gave a linear relation. These results suggest that anthocyanin is the major
source of antioxidant capacity in Roselle extract. Further purification using Amberlite XAD-2 and HPLC indicated that antho-
cyanin and a brown pigment in the extract account for about 51 and 24% of the antioxidant capacity, respectively. Under different
processing temperatures and storage periods, anthocyanin content declines. However, other phenolic compounds increase and
overall there is only a relatively small decrease in total phenolic compounds and antioxidant activity. # 2002 Elsevier Science Ltd.
All rights reserved.
Keywords: Anthocyanin; Antioxidant capacity; Roselle (Hibiscus sabdariffa L.)
Fig. 1. Effect of (a) extraction time and (b) sample weight on the FRAP of Roselle petals.
P.-J. Tsai et al. / Food Research International 35 (2002) 351–356 353
Table 1
Distribution of anthocyanin and antioxidant activity in Roselle infusion
The extract was passed through an Amberlite XAD-2 tion with Ciocalteu’s reagent. The amount of phenolic
column where the pigments were adsorbed. The column compounds in the Roselle extract (23 mg/g dry weight)
was initially washed with water to give the non-absor- is similar to that found by Mazza, Fukumoto, Delaquis,
bed fraction ‘‘A’’. This fraction contained 70% (by Givard, and Ewert (1999) in various fruits such as
weight) of the extracted material but only 3% of the strawberries and currants.
antioxidant activity. The pigments were eluted with The effects on composition of drying and storage at
methanol, giving Fraction ‘‘B’’ which contained 86% of different temperatures were investigated. After drying
the total pigments as estimated by A520. This fraction was and storage at 20 C for 15 weeks, 90% of the total
further separated into three sub-fractions on a LiChro- phenolic compounds remained. Storage at 40 C only
prep RP-8 column with an acetic acid: acetonitrile gra- decreased the phenolic compounds by a few percent.
dient: sub-fraction B1 was red, B2 was pink and B3 was Even after drying at 75 C and storage for 15 weeks at
brown. These data are summarised in Table 1 and Fig. 6. 40 C, 85% of the total phenolics remained. The effects
After comparison with authentic standards (pur- on the percentage composition of drying and storage at
chased from Polyphenols Laboratories, Sandnes, Nor- different temperatures are summarised in Table 2.
way) in the HPLC system, the red pigment was However, under these conditions, the anthocyanins
identified as delphinidin 3-sambubioside (85% of the decreased from about 80% of the total phenolics to
anthocyanin) and the pink pigment was provisionally about 50%. There was a corresponding increase in the
identified as cyanidin 3-sambubioside (Tsai & Ou, percentage of the other phenolics (Fig. 7). These appar-
1996). These two pigments account for 51% of the total ently contradictory results are consistent with the con-
FRAP activity. The remainder of the activity (24%), version of some monomeric anthocyanins into
was due to the brown pigment which was composed of polymerised phenolics during storage.
phenolic compounds. This conclusion is based on the Measurement of antioxidant activity under these
absorbance spectra of the brown fraction and its reac- conditions showed a strong correlation to anthocyanin
P.-J. Tsai et al. / Food Research International 35 (2002) 351–356 355
Table 2
Effect of drying and storage temperatures on the percentage of total phenolic compounds found in Roselle
Drying temp Storage temp Storage time GAE CCE CFAE RUE DEL Total phenolics
( C) ( C) (weeks) (%) (%) (%) (%) (%) (%)
Acknowledgements
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