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Materials and Methods process was conducted under light-

protected conditions.
Preparation of Crude Extract from
Butterfly Pea Flowers
Butterfly pea petals were air-dried in
the shade and ground into a fine
powder. The powder was gradually
dissolved in distilled water at a ratio
1:10 at room temperature. The crude
extract was collected by initially
filtering with gauzes and subsequently
with No. 1 Whatman grade filter
papers (Whatman, 1001-150). The pH
of the crude extract was measured
using a CyberScan 1000 pH meter
(Eutech Instruments Pte Ltd,
Preparation of Butterfly Pea Dye
for Staining
The filtered crude extract of butterfly
pea petals was added to 1.0%
aluminium chloride anhydrous and
1.2% Iron (III) chloride hexahydrate.
The dye solution wasmixed well and
filtered using No. 1 Whatman grade
filter papers. The pH was adjusted to
0.2 using concentrated HCl.
Staining on Blood Smear
The staining process was designed to
be conducted on blood smears of
chicken, pigeon, dog, and horse. The
smears were prepared from EDTA-
blood on glass slides. The slides were
air-dried and fixed in Diff-quick fixative
reagent which contained methanol
and triarylmethane dye for 30-60 sec.
The slides were stained with the
butterfly pea dye for 30 min. The
staining process was stopped by
covering the slides with glass covers
without washing and counter-staining.
Images of the stained smears were
taken under an Axiolab microscope
using Axio Vision software. All of the
Structural Identification and Mature Basella alba L. fruit, with dark blue
Bioactivities of Red-Violet Pigments skin and deep red-violet flesh, is a
Present in Basella alba Fruits potential source of natural colorants. Its
pigment components and bioactivities
deserve particular attention and
investigation. In this study, fruit flesh was
extracted with 80% methanol (containing
0.2% formic acid) and subjected to solid-
phase extraction, semipreparative HPLC
isolation, mass spectrophotometric
analysis, and structural elucidation. The
major red pigment was identified as
gomphrenin I. Its quantity increased with
the increase of fruit maturity. The
gomphrenin I extract yield from ripe fruits
was 36.1 mg/100 g of fresh weight. In
addition to gomphrenin I, betanidin-
dihexose and isobetanidin-dihexose were
also detected. The antioxidant activities of
gomphrenin I determined by Trolox
equivalent antioxidant capacity (TEAC),
,-diphenyl--picrylhydrazyl (DPPH)
radical scavenging activity, reducing
power, and antioxidative capacity assays
were equivalent to 534 M Trolox, 103 M
butylated hydroxytoluene (BHT), 129 M
ascorbic acid, and 68 M BHT at 180, 23,
45, and 181 M, respectively. The anti-
inflammatory function was tested at
concentrations of 25, 50, and 100 M in
murine macrophages stimulated with
lipopolysaccharide (LPS). The results
revealed that gomphrenin I suppressed
LPS-induced nitric oxide (NO) production in
a dose-dependent manner and decreased
PGE2 and IL-1 secretions at the highest
concentration tested. The transcriptional
inhibitory activities of gomphrenin I on the
expression of inflammatory genes
encoding iNOS, COX-2, IL-1, TNF-, and
IL-6 were also observed. It is of merit to
identify gomphrenin I as a principal
pigment of B. alba fruits and as a potent
antioxidant and inflammatory inhibitor.
These findings suggest that B. alba fruit is
a rich source of betalains and has value-
added potential for use in the
development of food colorants and
Spinach vine extract was more stable
at pH 5.0 and 6.0 than at pH 4.0 both
in the presence and absence of light.
This characteristic differs from other
anthocyanins. This property could
facilitate its application as a natural
Stability of anthocyanin in food colorant.
spinach vine (Basella rubra)
fruits. Cien. Inv. Agr. (In English)
E. Ferreira Ozela, Paulo Csar icle/view/389
Stringheta, Milton Cano Chauca


The stability of anthocyanin in the Utilization of an indigenous

extract of spinach vine fruit (Basella TITLE dyestuff from Basella rubra
rubra L.) was studied in relation to (alugbati) as microbiological stain
degradative factors such as light, CALL NO(S) Fil Q149.P5 N25
temperature and pH acting alone or in
combination. In this work, the possible
use of spinach vine fruit as a source of PUBLICATION Transactions of the National
TITLE Academy of Science and Technology
natural pigments for use in food
coloring was evaluated. Extraction of VOLUME/ISSUE 28(1)
the pigment was carried out with ISSUE DATE 2006
99.9% methanol at pH 2.0. The
MAIN AUTHOR Enerva, Lorna T. , et al
stability of the anthocyanin extract
was estimated. From these values, ABSTRACT This study was undertaken to
extract anthocyanin from Basella
reaction velocity constants (k) as well rubra berries and utilize the extract
as the half-life time (t1/2) were as microbiological stain. It is an
calculated at pH 4.0, 5.0, and 6.0 in inexpensive, indigeneous and
abundant raw material. The alugbati
the presence and absence of light
berries were macerated in a blender
both at 40 and 60C. Results indicate and extracted with 1% HCl in 95%
that, independent of pH values, methanol. The extract obtained was
spinach vine extract suffered an filtered and then concentrated. Thin
layer and column chromatography
interference of light in its anthocyanin methods were used to isolate and
degradation kinetics with the mean purify the anthocyamin. The samples
t1/2 being greater in samples place in were analyzed using infrared spectra
and ultraviolet spectra. FT-IR revealed
darkness (654.5 66.6 h) compared the presence of a hydroxyl group
to exposed to light (280 60.62 h). In which is prominent in the structure of
the presence of light, degradation of anthocyanin pigment at 3385cm'1. The
the anthocyanin pigment increased C=O stretching for aromatic ring were
indicated by a peak at 1635.20 cm'l,
with increased temperature and had 1513.38 em-I for C=O bending,
an average half life time of 280 1439.91 cm'I for O-H bending, 1207.42
60.62 h, 6.88 0.76 h and 2.42 cm,l for c-o stretching, 1151.92cm"
for alkane and 1100.77 em-I for C-C
0.31 h at room temperature (25 stretching. The structure of
1C), 40 and 60C, respectively. anthocycanin was further established
by the e maximum of the ultraviolet structure of the microorganisms with
spectrum at 510.0 nm. For the respect to shape and size and certain
application, the crude extract was cellular components were identified
used as a stain for Staphyloccus using a microscope and
aureus, a gram positive bacteria and photomicrographs. The alugbati
Escherichia coli, a gram negative extract produced stain that was
bacteria. The staining process for the comparable with synthetic stains like
microorganism used mordants like crystal violet and safranin and can,
potassium alum, calcium oxide and therefore, be used as an alternative
copper sulfate for fixing the color. stain.
Only copper sulfate and lime
responded positively as a mordant SUBJECTS Biological sciences
that gave favorable outcome in fixing Basella rubra (alugbati)
the color of alugbati. The samples Microbiological stain
were screened based on the criteria
of color retention and evenness. The