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Nhận Biết Porphyrin PDF
Nhận Biết Porphyrin PDF
To elucidate the substance which causes fluorescence in black-lip pearl, the substance was extracted
from black-lip pearls with 2N-HCl. After being fractionated by talc column chromatography , the
substance was converted to methyl ester. Using uroporphyrin I octamethyl ester as a standard, the
esterified substance was compared in terms of visible absorption spectra , fluorescence emission spectra,
and high performance liquid chromatographic behaviors. Considerable similarities in absorption peaks,
fluorescence peak, and retention time resulted between the esterified substance and the standard. From
these results the substance was proved to be uroporphyrin I. The experimental result of the existence
of porphyrin facilitates percent pearl identification.
Key words: porphyrin pigment, black-lip pearl, black-lip oyster, pearl identification
Black pearls which are cultured using black-lip oyster Extraction of Pigments
Pinctada margaritifera are highly evaluated because of their Two hundred grams each of the nacre of both pearl and shell were
milled. Pigments were extracted by decalcification with 2N-HCl. By
high quality and scarcity. When black-lip pearls first filtration with filter paper (Advantec No. 2) the residue (conchiolin) was
appeared on the market in the 1970's, there were may claims removed. Extraction was repeated until the extract became colorless. The
that they had been chemically treated or dyed, since many extracts were combined and concentration under a vacuum.
tinged black pearls, most of which were cheaper akoya
Pinctada fucata pearls had already been sold. Therefore the Fractionation of Pigments
The fractionation of the pigment was performed by passing the
identification between these two different kinds of black concentrated extract through a talc column (1.8 •~ 7 cm). Developed bands
pearls is of much importance. Actually, black pearl were washed by 100 ml of water and then eluted by a subsequent flow of
identification is based on detecting the characteristics of 1% sodium acetate (100 ml) and 10% aqueous ammonia (50ml). The
black-lip pearls. For example, the black color of black-lip fraction eluted by 10% aqueous ammonia fluoresced as a vivid red color,
and was used for analysis.
pearls was examined using infrared color film.1) By tone
differences on the film, black-lip pearls were differentiated Esteriftcation of Pigment
from tinged black ones. At present, though the reason is After being evaporated to dryness under the vacuum, the ammonia
not still clear, the fact that black-lip pearls have a specific eluted fraction was dissolved in a mixture of sulfuric acid and methyl
reflectance spectrum near 700 nm is commonly used for alcohol (2:8, v/v) and was left for 16 h under 10-15•Ž. The methyl ester
of the pigment was extracted with chloroform. The chloroform layer was
identification.2) Red fluorescence is another effective method
washed with water, 0.5% sodium bicarbonate, and water successively, and
for pearl identification: black-lip pearls possess a specific was then dehydrated on anhydrous sodium sulfate.
peak fluorescence emission at 620 nm with 400 nm radia
tion. Miyoshi et al.3) examined the fluorescence of sev Spectrometry
eral pearl-producing oysters and reported that the red Visible absorption spectra of the methyl ester of pigments were measured
in chloroform between 300 and 700 nm with a Nihon Bunko UVIDEC
fluorescence of black-lip oyster and mabe oyster Pteria 610C spectrophotometer. Fluorescence emission spectra were also mea
penguin was derived from porphyrin. This result suggests sured with a Nihon Bunko FP 770 spectrophotofluorometer. Uropor
that the red fluorescence in black pearls produced by phyrin I octamethyl ester (Sigma) was used as a standard. The fluores
black-lip oysters is derived from uroporphyrin. Our cence emission spectra of both black-lip pearls and shell were measured
investigation was undertaken on the porphyrins both in non-destructively with a Nihon Bunko FP 770 spectro-photofluorom
eter.
black-lip oysters and pearls. Results obtained showed that
uroporphyrin exists in the nacre of both pearls and shells. High-Performance Liquid Chromatography
Methyl ester of the pigments and uroporphyrin I octamethyl ester were
Materials and Methods subjected to HPLC4) on a column of Shim-pack CLC-SIL (M) (4.6 •~
250 mm) with a mobile phase of n-hexane-ethyl acetate-dichloromethane
Fig. 1. Absorption spectra of methyl ester of pigments from black-lip pearl (a) , black-lip shell (b), and uroporphyrin I octamethyl ester (c).
References
and the same non-destructive spectroscopic measurement 13) S. Akamatsu, H. Komatsu, C. Koizumi, and J. Nonaka: A
comparison of sugar compositions of yellow and white pearls. Nippon
described above showed no evidence of porphyrin. There
Suisan Gakkaishi, 43, 773-777 (1977).
fore porphyrin is one of the characteristics of black-lip
14) A. Comfort: Acid soluble pigments of shells. The distribution of
pearls. Our experimental result can facilitate the present porphyrin fluorescence in molluscan shells. Biochem. J., 44, 111-117
black pearl identification with fluorescence emission spectra (1949).
in terms of porphyrin.