Few studies have been reported dealing with the influence of anthocyanins on the growth of bacteria. Blank and Suter (3) investigated the effect of anthocyanins on pathogenic bacteria and reported that concentrations of 0.02 and 0.1M had no bactericidal action. The work of Blank and Suter was substantiated by Mandrik (7) who also reported that the anthocyanins had no apparent effect on pathogenic bacteria.Masquelier and Jensen (9, lo), in studying the bactericidal action of red wine, reported that pigments isolated from grapes to be used in red wine were not bactericidal. On hydrolysis, a fraction containing partially demethoxylated oenidol had definite bactericidal activity. Masquelier and Jensen found that as wine ages oenoside (malvidin 3-monoglucoside) was slowly converted to oenidol (malvidin). The oenidol thus formed possessed a phenol coefficient of 33. Masquelier and Jensen thought this might explain the bactericidal action of red wine.Zimmerman (1 7) found that strawberry and grape anthocyanins significantly influenced the growth rates of Lactobacillus acidophilus and Cameron's Putrefactive Anaerobe No 3679 (P.A. 3679). Zimmerman found that the growth rate of P.A. 3679 was stimulated by anthocyanins. The growth of L. acidophilits was stimulated by 0.008 to 0.025 mg % pigment while 2.5 mg % depressed the growth rate. H e found no difference in the effect of strawberry and grape anthocyanins. EXPERIMENTAL PROCEDURE Isolation of anthocyaninsThe anthocyanins investigated in the present study were isolated from comniercially frozen strawberries and commercially frozen and bottled Concord grape juice. Frozen strawberries were macerated in a Waring blender and the pulp removed from the juice by pressing through three layers of cheese cloth.Extrnctiou: The pigments were extracted from the fruit juice with acidified (1% HCI) 1-butanol. Two liters of the acidified butanol were used per liter of fruit juice. The aqueous and organic phases were separated and extraction of the pigments repeated. The pigments were precipitated into an aqueous phase by the addition of petroleum ether to the pigment-butanol solution. The organic phase was discarded.Chronzofogrnphic solvents: Two solvents were prepared by thoroughly shaking 1-butanol, acetic acid, and water (4: 1 : 5 v/v) in a separatory funnel. On standing the mixture separated into 2 phases. Herein, the upper phase (water-saturated butanol) is referred to as solvent A and the lower phase (butanol-saturated water) as solvent B (14). Chromatograpliic separation: The aqueous pigment extracts were concentrated in VUCTCO and heavily streaked on rectangular (8 x 12 in) Whatman No 1 filter paper. Using solvent A and the ascending chromatographic technique the chromatograms were de-

SUMMARY The metal‐complexing properties of malvidin‐3‐monoglucoside, petunidin‐3‐monoglucoside, and 3′‐methoxy‐apigeninidin‐chloride‐4′‐methyl‐ether were studied polarographically and spectrophotometrically to complement bacteriological findings previously made. The molar ratios at which malvidin‐3‐ monoglucoside combined with cupric ions depended on the pH of the supporting electrolyte solutions. The molar ratio of cupric ion to malvidin‐3‐monoglucoside was 2:1 at pH 3.9; the molar ratios were 1:2 at pH 3.5 and 4.5, but these ratios were 1:1 at pH 5.0 and 6.0. The absorption peaks of malvidin‐3‐monoglucoside were shifted to shorter wavelengths in the visible range but to longer wavelengths in the UV range upon addition of various concentrations of cupric ions. The absorption peaks of petunidin‐3‐monoglucoside were shifted to shorter wavelength in both the UV and the visible light ranges upon addition of various concentrations of cupric ions. The infrared absorbancies of 3′‐metboxy‐apigeninidin‐chloride‐4′‐methyl‐ether at 11 to 14.5 μ disappeared upon the addition of Ca and Mg ions.

Six anthocyaninswere isolated from Cabernet Sauvignon grapes. Four major and one minor pigment were identified by paper chromatography, color reactions, and spectroscopy.The four major anthocyanins were identified as delphinidin-3-monoglucoside, petunidin-3-monoglucoside, malvidin-3monoglncoside, and malvidin-3-monoglueoside aeetylated with chlorogenic acid. Malvidin-3-monoglucoside was the most abundant pigment of the grapes. One of the minor pigments was identified as petnnidin.

SUMMARY Pelargonidin 3‐monoglucoside was isolated from frozen strawberries and subjected to heat degradation in IN HCl. The six fractions isolated were characterized chemically and physically. The unheated and heated pigment inhibited the growth of Escherichia coli and Staphylococcus aureus, and exerted both a stimulatory and inhibitory effect on Lactobacillus casei culture. The stimulation may be due to a decrease in the oxidation‐reduction potential of the media affected by the pigment, and/or the ability of the organism to split the β‐glyeosyl bond and use the glucose moiety. The influence on growth of the test organisms of the various fractions resulting from heating the pigment was reported.

Various objective methods to estimate the degree of freshness of fish and fishery products were compared. The total volatile nitrogen (TVN) and ammonia-nitrogen of carp flesh reached values of 30–33 mg% when carp stored at 0.4°C. and 25–27°C. began to have a bad odor. For catfish stored at 0–4°C. and 25–27°C. these values reached 20–30 mg%. Trimethylamine was not found in fresh and spoiled carp and catfish. The determination of trimethylamine as an index of spoilage cannot be used for these fresh-water fish. The determinations of tyrosine and volatile reducing substances (VRS) cannot be used to estimate the degree of freshness of fish if the temperature at which the fish had been stored is not known. If these two methods were combined with TVN and ammonia-nitrogen one could determine the degree of freshness of fish and the temperature at which the fish were stored. The determination of tyrosine combined with TVN and ammonia-nitrogen can give further support as to whether fish are absolutely fresh. If the temperature under which fish have been stored is known each of the tests used in this study can be applied individually to determine the degree of freshness of fish.