Materials and Methods process was conducted under light-

protected conditions.
Preparation of Crude Extract from
Butterfly Pea Flowers http://www.thaiscience.info/journals/Ar
ticle/SJST/10890421.pdf
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,
Singapore).
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
Bioactivities of Red-Violet Pigments
Present in Basella alba Fruits
Mature Basella alba L. fruit, with dark blue
skin and deep red-violet flesh, is a
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
nutraceuticals.
http://pubs.acs.org/doi/abs/10.1021/jf1
017719

Stability of anthocyanin in
spinach vine (Basella rubra)
fruits. Cien. Inv. Agr. (In English)
34(2):85-90.
E. Ferreira Ozela, Paulo Csar
Stringheta, Milton Cano Chauca
Abstract
The stability of anthocyanin in the
extract of spinach vine fruit (Basella
rubra L.) was studied in relation to
degradative factors such as light,
temperature and pH acting alone or in
combination. In this work, the possible
use of spinach vine fruit as a source of
natural pigments for use in food
coloring was evaluated. Extraction of
the pigment was carried out with
99.9% methanol at pH 2.0. The
stability of the anthocyanin extract
was estimated. From these values,
reaction velocity constants (k) as well
as the half-life time (t1/2) were
calculated at pH 4.0, 5.0, and 6.0 in
the presence and absence of light
both at 40 and 60C. Results indicate
that, independent of pH values,
spinach vine extract suffered an
interference of light in its anthocyanin
degradation kinetics with the mean
t1/2 being greater in samples place in
darkness (654.5 66.6 h) compared
to exposed to light (280 60.62 h). In
the presence of light, degradation of
the anthocyanin pigment increased
with increased temperature and had
an average half life time of 280
60.62 h, 6.88 0.76 h and 2.42
0.31 h at room temperature (25
1C), 40 and 60C, respectively.
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
food colorant.
http://www.rcia.uc.cl/index.php/rcia/art
icle/view/389
Utilization of an indigenous
TITLE dyestuff from Basella rubra
(alugbati) as microbiological stain
CALL NO(S) Fil Q149.P5 N25
LOCATION(S) STII
PUBLICATION Transactions of the National
TITLE Academy of Science and Technology
VOLUME/ISSUE 28(1)
ISSUE DATE 2006
MAIN AUTHOR Enerva, Lorna T. , et al
ABSTRACT This study was undertaken to
extract anthocyanin from Basella
rubra berries and utilize the extract
as microbiological stain. It is an
inexpensive, indigeneous and
abundant raw material. The alugbati
berries were macerated in a blender
and extracted with 1% HCl in 95%
methanol. The extract obtained was
filtered and then concentrated. Thin
layer and column chromatography
methods were used to isolate and
purify the anthocyamin. The samples
were analyzed using infrared spectra
and ultraviolet spectra. FT-IR revealed
the presence of a hydroxyl group
which is prominent in the structure of
anthocyanin pigment at 3385cm'1. The
C=O stretching for aromatic ring were
indicated by a peak at 1635.20 cm'l,
1513.38 em-I for C=O bending,
1439.91 cm'I for O-H bending, 1207.42
cm,l for c-o stretching, 1151.92cm"
for alkane and 1100.77 em-I for C-C
stretching. The structure of
anthocycanin was further established
by the e maximum of the ultraviolet
spectrum at 510.0 nm. For the
application, the crude extract was
used as a stain for Staphyloccus
aureus, a gram positive bacteria and
Escherichia coli, a gram negative
bacteria. The staining process for the
microorganism used mordants like
potassium alum, calcium oxide and
copper sulfate for fixing the color.
Only copper sulfate and lime
responded positively as a mordant
that gave favorable outcome in fixing
the color of alugbati. The samples
were screened based on the criteria
of color retention and evenness. The
structure of the microorganisms with
respect to shape and size and certain
cellular components were identified
using a microscope and
photomicrographs. The alugbati
extract produced stain that was
comparable with synthetic stains like
crystal violet and safranin and can,
therefore, be used as an alternative
stain.
SUBJECTS Biological sciences
Basella rubra (alugbati)
Microbiological stain