Food Chemistry I1 (1983) 167-174

Changes in the Chemical Constituents of

New Zealand Grapefruit During Maturation

Gordon L. Robertson
Department of Food Technology, Massey University,
Palmerston North, New Zealand

&
Myrna O. Nisperos
Department of Food Science and Technology, University of the Philippines at Los Banos,
College, Laguna 3720, Philippines

(Received: 18 August, 1982)

ABSTRACT

New Zealand grapefruit were harvested at intervals throughout the 1978

and 1981 seasons and their juices analysed for several chemical
constituents. The pH ranged from 3"0 to 3.8, the total soluble solids from
10.3 to 14.4 °Brix, the titratable acid from 0.92 to 2"56g citric'
acid/lOOml, the Brix.'acid ratio from 4"2 to 14.1, and the ascorbic acid
from 33"2 to 38.3mg/lOOml. Total flavonoids ranged from 626 to
1470 mg/litre, naringin from 528 to 743 mg/litre and limonin from 4.7 to
20.2 mg/litre. There was considerable variation in the concentration of
the bitter constituents between the two seasons. The results obtained are
compared with other published figures and their implications for juice
processors and consumers are discussed.

INTRODUCTION

Although N e w Zealand is situated climatically at the very southern limit

for citrus culture, the citrus industry has expanded steadily over the past
167
Food Chemistry 0308-8146/83/$03"00 © Applied Science Publishers Ltd, England, 1983.
Printed in Great Britain
168 Gordon L. Robertson, Myrna O. Nisperos

decade with the total citrus crop doubling from 14045 tonnes in 1974 to
28 647 tonnes in 1980 (Anon., 1981). While only 20 ~o of the citrus crop is
processed, New Zealand grapefruit (hereafter referred to as NZGF)
accounts for over half of this tonnage, making it the most important
citrus fruit from a processor's point of view.
N Z G F is a natural hybrid of obscure origin with tangelo characteristics
(Hodgson, 1967). Although limited information is available on the
composition o f N Z G F juices (Dawes, 1970; Robertson, 1975), there have
been no published reports on the nature of the bitter compounds in
N Z G F or the way in which their concentration might change during
maturation. The study reported here was undertaken to provide such
information.
Bitterness in citrus fruits and products is primarily caused by two
groups of compounds--flavonoids and limonoids (Kefford & Chandler,
1970). Flavonoid bitterness is due mainly to naringin and has been
studied most frequently with respect to grapefruit products. Naringin
occurs in low concentrations in the juice of mature grapefruit, and in
higher concentration in segment membranes, core and peel (Sinclair,
1972).
Limonoid bitterness is associated with Navel oranges and grapefruit.
Limonin, the major limonoid, causes delayed bitterness in the juice from
early season to mid-season Washington Navel, Australia Valencia and
Israel Shamouti oranges, as well as from Marsh grapefruit. Limonoid
bitter principles generally decrease with maturity of the fruit (Maier et al.,
1977).

EXPERIMENTAL

Raw material

The N Z G F used in this study were of the Golden Special variety

propagated on trifoliata (Poncirus trifoliata) rootstock in the Massey
University orchard, Palmerston North. The trees were planted in 1975.
During both the 1978 and 1981 seasons, six trees were selected at random
from the orchard and at intervals from early June or July until late
October, two fruit were picked from each tree. The selected fruits were
judged to be of average size and colour for the tree at that harvest and
picking was evenly distributed around the outside of the trees.
 Changes in the chemical constituents of New Zealand grapefruit 169

Juice was extracted from the fruit with a mechanical hand-reamer and
screened through a double layer of cheese-cloth to remove gross
suspended matter. Where necessary juice was pasteurised by heating to,
and maintaining, a temperature of 95 °C for 5 min and then cooling.

Common juice parameters

The pH was determined using a combination electrode connected to a

Triac pH meter with digital output. Prior to measurements being made
the instrument was calibrated with buffers at pH 3.2 and 7.0. Total
soluble solids (TSS) were measured by refractometer and expressed as
degrees Brix (°B). Titratable acidity was determined by titration of the
juice against N a O H (0.1 ~) using phenolphthalein (0"5~oW/V) as an
indicator. Results are expressed as grams of citric acid per 100 ml of juice.
Ascorbic acid was determined spectrophotometrically using 2,6 dichloro-
phenolindophenol (Pearson, 1976).

Bitter compounds

Total flavonoid content was determined during the 1978 season using the
Davis test (Davis, 1947). It has been shown by Hagen et al. (1966) and
Tatum et al. (1972) that the total flavonoid content, as determined by the
Davis test, is approximately 2"1 times the naringin concentration.
During the 1981 season the naringin concentration was determined
using the thin layer chromatographic-spectrophotometric method of
Tatum & Berry (1973a).
Limonin was analysed by a method involving thin layer chroma-
tography (TLC) and densitometry which combined, with slight modifi-
cations, the separation procedure of Chandler (1971) and the visualisation
procedure of Tatum & Berry (1973b). Quantitation was based on
measurement of the heights of densitometric peaks corresponding to the
standard and unknown limonin spots on the same TLC plate using a
Shimadzu CS-910 TLC scanner. This analytical procedure does not
distinguish between limonin and its non-bitter precursor which is
generally believed to be limonoic acid A-ring lactone (Maier et al., 1977).
The precursor is extracted from the juice and converted into limonin
during preparation of the extract for chromatography.
170 Gordon L. Robertson, Myrna O. Nisperos

RESULTS AND DISCUSSION

The changes in the chemical constituents of NZGF juice during

maturation are summarised in Table 1. With the exception of ascorbic
acid, all the parameters measured changed significantly during maturation.
The increases in pH and TSS and decreases in titratable acidity parallel
very closely the trends reported by others (Sinclair, 1972; Robertson,
1975).
The ascorbic acid concentration reported here is within the range of 25
to 60 mg/100 ml reported by Nagy (1980) for a wide variety of grapefruit
grown in various parts of the world. The figures are marginally but
significantly higher than those reported earlier by Robertson (1975) for
NZGF juice (mean value of 29.0mg/100ml), possibly because the juice
analysed in this present study was extracted immediately after harvesting
compared with a delay of 3 days in the earlier study. Recently, Albach
et al. (1981a) reported constant values of ascorbic acid in early and mid-
season grapefruit, a trend which is evident in the results in Table 1.
Albach also reported a 3-year mean of ascorbic acid in grapefruit juice
(primarily the Ruby Red variety) of 31.3 mg/100 ml; this compares with
36.9 in the present study.
Care is needed when comparing the naringin values in Table 1 with
other published results since many authors erroneously equate the total
flavonoid concentration as determined by the Davis method with
naringin concentration. Fisher (1976) developed a far more specific
HPLC procedure for determining naringin and this method was used by
Dougherty & Fisher (1977) and Ting & McAllister (1977). They
determined the naringin contents of Florida grapefruit juice and reported
ranges of 152-711,240-903 and 123-442mg/litre and averages of 310,
456 and 285 mg/litre respectively. Comparison of these values with those
presented in Table 1 indicates that NZGF juices contain significantly
higher concentrations of naringin (1981 average 650mg/litre) than do
Florida grapefruit juices. On the other hand, the total flavonoid contents
of NZGF juice (1978 average 1153 mg/litre) fall within the range of values
reported by Maurer et al. (1950) but outside that reported by Dougherty
& Fisher (1977) for USA grapefruit juice; namely, 130-1880 and
277-910 mg/litre respectively.
Although the data are not strictly comparable, it is clear that flavonoid
contents were higher in 1981 than in 1978. Also, there was a more than
doubling of concentration of total flavonoids in 1978 compared with a
 TABLE 1
Changes in the Chemical Constituents of NZ Grapefruit Juice During Maturation

Pick ing pH Total Titratable Brix: acid Ascorbic Total Naringin Limonin
date soluble acidity ratio acid flaeonoids (mg/litre) (mg[litre)
solids (gcitric (mg/lOOml) (mg/litre) ~a
(°Brix) acM/lOOml)

11 July 1978 3.l 10.3 1.83 5.7 ND a 626 ND 17.9 ~.

2 August 1978 3.4 11.6 1.47 7.9 ND 980 ND 18.6 e~
22 August 1978 3.6 12.2 1.08 11.3 ND 1 060 ND 17-4
13 September 1978 3"8 12.6 0.94 13.4 ND 1 420 ND 20.2 ~.
3 October 1978 3.7 12.1 0.96 12.6 ND 1 470 ND 12.4 ~"
25 October 1978 3.7 12.9 0.92 14' 1 ND 1 362 ND 11 "8

2 June 1981 3.0 10.8 2.56 4.2 ND ND 743 7-5

22 June 1981 3.1 11.6 2-40 4.8 37.8 ND 615 8-8
13 July 1981 3.1 12.1 2.12 5.7 36-5 ND 575 6.1 ~N
3 August 1981 3.2 12.8 2-08 6.2 37.8 ND 655 5.7 ~-
24 August 1981 3.3 13.6 1.86 7.3 38.3 ND 662 4.8
24 September 1981 3.4 14.4 1.84 7"8 37.8 ND 528 4.8
5 October 1981 3.4 13.5 1.63 8.3 37.3 ND 670 5-3 "~
26 October 1981 3"5 13.5 1.31 10.3 33.2 ND 735 4-7 ~'~

Not determined.
172 Gordon L. Robertson, Myrna O. Nisperos

relatively constant concentration of naringin in 1981 ; these two processes

are not mutually exclusive. The steady increase in total flavonoid
concentration during a season does not appear to have been observed
previously, although the reverse does: Maurer et al. (1950) reported a
decrease in juice naringin concentration as the season progressed with a
slight rise towards the end of the season. In contrast, Kesterson &
Hendrickson (1953) and Albach et al. (1981b) observed no significant
decrease in juice naringin concentration until late in the season.
The actual concentrations of limonin are quite different in the two
seasons, with values in 1978 about three times those in 1981. Moreover, for
the 1978 season the changes in limonin content are rather abrupt with an
almost constant level of limonin for 2 months, followed by a sharp drop
over a period when the 1981 values remain relatively constant. It has been
well established that the limonin content of citrus fruit tissues decreases
with advancing maturity (Chandler et al., 1976; Albach et al., 1981 a). For
example, Chandler et al. reported that the limonin content in the juice of
Navel oranges grown on trifoliata rootstock fell approximately from
22 ppm on 24 April to 12 ppm on 25 June to 6 ppm on 25 August. From
published data it appears that rates of disappearance of limonin vary
markedly with the crop for both horticultural and climatic reasons. For
example, Mansell & Mclntosh (1980) reported that there was no
relationship between limonin, Brix, acid and Brix: acid ratio in grapefruit
juice samples from three State Test Houses in Florida.
The Florida Department of Citrus (Anon., 1975) has specified that a
Grade A grapefruit juice, processed outside the period for peak quality,
must contain less than either 5 mg/litre limonin or 600 mg/litre naringin.
European workers (Buffa & Bellenot, 1962) have suggested that a
naringin content of 300-700 mg/litre is necessary for the characteristic
grapefruit flavour, otherwise the juice is too insipid or too bitter. In both
of the above cases naringin content is as determined by the Davis test and
is thus strictly speaking the total flavonoid content.
While these figures can only be viewed as guidelines because of the
recognised effects of acidity and sweetness on subjective responses to
bitterness (Kamen et al., 1961), they do suggest that none of the juice
extracted from N Z G F during the 1978 or 1981 seasons could be regarded
as acceptable by consumers. Citrus processors in New Zealand have
recognised this fact for many years and marketed N Z G F juice either in
combination with other fruit juices or diluted with sugar and water as a
fruit drink.
 Changes in the chemical constituents of New Zealand grapefruit 173

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