Chlorophyllins

Chiorophyllin—copper complex, oil soluble—The chlorophyllin is obtained by extraction from a mixture of fescue and rye grasses. The chlorophyll is... 

Potassium sodium copper chlorophyllin (chiorophyllin—copper complex)—A green-black powder obtained from chlorophyll by replacing the methyl and phytyl ester groups with alkaH and replacing the magnesium with copper. The source of the chlorophyll is dehydrated alfalfa. 

Hydrolysis of chlorophyll using sodium hydroxide produces the moderately water-soluble sodium salts of chlorophyllin, phytol and methanol (145). The magnesium ia chlorophyllin may be replaced by copper. The sodium copper chlorophyllin salt is heat stable, and is ideal for coloring foods where heat is iavolved, such as ia canning (146). 

In a clinical trial performed in China, the administration of 300 mg/day of copper chlorophyllin to humans who had detectable levels of serum aflatoxin due to unavoidable food contamination resnlted in a 50% reduction of median urinary levels of aflatoxin-DNA adducts. If health benefits from consuming natural chlorophylls were confirmed, it wonld be easy to add green leafy vegetables to a daily diet to obtain the benefit. Since leafy vegetables contain usually up to 200 mg chloro-phylls/100 g fresh weight, the intake of approximately 1 to 2 cups of raw spinach/day... 

Blum, C.A. et al., F romotion versus suppression of rat colon carcinogenesis by chlorophyllin and chlorophyll Modnlation of apoptosis, cell proliferation, and 3-catenin/Tcf signaling, Mutat. Res., 523, 217, 2003. 

Carter, O., Bailey, G.S., and Dashwood, R.H., The dietary phytochemical chlorophyllin alters E-cadherin and P-catenin expression in human colon cancer cells International Research Conference on Food, Nutrition and Cancer, J. Nutr., 134,3441S, 2004. 

Nelson R.L. Chlorophyllin, an antimutagen, acts as a tumor promoter in the rat dimethylhydrazine colon carcinogenesis model. Anticancer Res., 12, 737, 1992. 

Dashwood, R.H., The importance of using pure chemicals in (anti) mutagenicity studies Chlorophyllin as a case point, Mutat. Res., 381, 283, 1997. 

Commercially produced metal-substituted chlorophylls such as copper chlorophylls and copper chlorophyllins that can be obtained by chemical modification of natural chlorophylls have better stability, solubility, and tinctorial strength, but they cannot be considered natural food colorants and will be discussed later. 

Although chlorophyll and chlorophyllin colorants seem to be easily obtained, in practice their production as natural food colorants is rather difficult. The sensitivity of chlorophylls to certain enzymes, heat, and low pH, and their low tinctorial strength greatly limit their manufacture and application as food additives, principally when the pigments are isolated from the protective environment of the chloroplasts. The well-known instability of chlorophylls prompted extensive research for developing... 

In recent years, metallo-chlorophylls and metaUo-chlorophyllins have been considered alternatives to their natural chlorophyll counterparts due to their enhanced color potency, and greater stability against moderate heat, dilute acids, and oxidative agents in general, not to mention their alleged biological activities. 

Commercial food grade water-soluble Cu-chlorophyllin is the most notable among these preparations. Copper chlorophyllins are produced from crude natural chlorophyll extracts followed by the hydrolysis of the phytyl and methyl esters, cleavage of the cyclopentanone (E) ring in dilute alkali, and the replacement of magnesium by copper. ... 

Chemical analysis revealed that commercial food grade copper chlorophyllin is not a single, pure compound, but is a complex mixture of structurally distinct porphyrins, chlorin, and non-chlorin compounds with variable numbers of mono-, di-, and tri- carboxylic acid that may be present as either sodium or potassium salts. Although the composition of different chlorophyllin mixtures may vary, two compounds are commonly found in commercial chlorophyllin mixtures trisodium Cu (II) chlorin Cg and disodium Cu (II) chlorin which differ in the number of... 

It must be remembered that copper chlorophylls and copper chlorophyllins are chemically modified natural extracts and therefore should not be called natural. 

Fermzzi, M.G. and Schwartz, S.J., Thermal degradation of commercial grade sodium copper chlorophyllin, J. Agric. Food Chem., 53, 7098, 2005. 

Scotter, M.J., Castle, L., and Roberts, D., Method development and HPLC analysis of retail foods and beverages for copper chlorophyll (E 141 [i]) and chlorophyllin (E 141[ii]) food colouring materials, FoodAddit. Contam., 22, 1163, 2005. 

U.S. Eood and Drug Administration, Listing of color additives exempt from certification sodium copper chlorophyllin, 21CER Part 73, 67 Fed. Reg. 35429, May 20, 2002. 

In contrast to the well-established methods for identifying and quantifying naturally occurring chlorophylls, very few reports concern quantitative analysis of chlorophyllin copper complexes in color additives and in foodstuffs. Analytical methods proposed are based on spectral properties, elemental analysis, chromatographic separation, and molecular structure elucidation or a combination of these procedures. 

Sodium copper chlorophyllin, approved by the FDA as a color additive in citrus-based dry beverage mixes, should have a ratio of absorbance (SoretQ band) not less than 3.4 and not more than 3.9. In Europe, purity criteria of the food additives E141[i] and E141[ii], which are copper complexes of chlorophyll and chlorophyllin, respectively, are set out in the EC color specifications that include identification and spectrophotometric assay tests. ... 

Purity levels of commercial preparations have also been estimated based on their Cu contents and compared with the theoretical values expected for fuUy coppered chlorophyllin based on the two major compounds Cu(ll) chlorin e4 (disodium salt) and Cu(ll) chlorin e6 (trisodium salt). The expected theoretical content of copper in a pure Cu chlorophyllin complex is 9.2%, which has never been found in commercial preparations. The sodium copper chlorophyllin from Sigma-Aldrich (St. Louis, MO) has a 4.5% copper content, specified by the manufacturer with respect... 

Analysis of nitrogen contents could be an aid for estimating the chlorophyllin concentration, complementing the copper analysis. The Cu N ratio of 1.1 calculated based on a Cu content of 9.2% and N content of 8.1% must be re-evaluated because both Cu and N levels found in commercial preparations are significantly lower than theoretical values. 

Similarly to the methods used to characterize natural chlorophylls, RP-HPLC has been chosen by several authors to identify the individual components in Cn chlorophyllin preparations and in foods. The same ODS columns, mobile phase and ion pairing or ion suppressing techniques coupled to online photodiode UV-Vis and/or fluorescence detectors have been used. ° ... 

Inoue, H. et al., Determination of copper(II) chlorophyllin by reversed-phase high-performance liquid chromatography, J. Ghromatogr. A, 679, 99, 1994. 

Chernomorsky, S., Quantitative procedure for chlorophyllin copper complex. Technical Communications, J. AOAC Internal., 11, 765, 1994. 

Chernomorsky, S. et al.. Antimutagenicity, cytotoxicity and composition of chlorophyllin copper complex. Cancer Lett., 120, 141, 1997. 

Breinholt V, Hendricks J, Pereira C, Arbogast D and Bailey G. 1995. Dietary chlorophyllin is a potent inhibitor of aflatoxin Bi hepatocarcinogenesis in rainbow trout. Cancer Res 55(l) 57-62. 

Sarkar D, Sharma A and Talukder G. 1994. Chlorophyll and chlorophyllin as modifiers of genotoxic effects. Mutat Res 318(3) 239-247. 

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