Extraction of carotenoid-rich palm pressed fiber oil using mixtures of hydrocarbons and short chain alcohols.

Solvent extraction is the most efficient method for recovering residual oil from palm pressed fiber (PPFO), which may contain up to eight times the carotenoid content of that found in crude palm oil. The objective of the present study is the use of binary mixtures of hydrocarbons (HC), hexane (Hex), cyclohexane (CHex) or heptane (Hep), and alcohols (ALC), ethanol (Eth) or isopropanol (IPA), in order to promote the highest recovery of a carotenoid-rich PPFO, in which the compositions of the mixtures are defined based on the calculation of solute-solvent distance (Ra) considering ß-carotene as the solute. The extraction experiments were conducted in batch, at 60 ± 2 °C, or in a fixed-bed packed column, at 55 ± 3 °C. Hex and Hep:IPA provided 80% of batch PPFO extraction yield, while in column, the highest yields were obtained with Eth and Hex:IPA (66%). The total carotenoid content obtained was the same independent of the solvent and extraction configuration (from 1790 ± 230 up to 2539 ± 78 mg ß-carotene/kg PPFO). In terms of the carotenoid profile, ß-carotene was mostly extracted by Hex, Hex:Eth stood out in the extraction of α-carotene, and Eth extracted the highest content of lycopene. It is possible to infer that mixtures of HC and ALC with compositions defined based on Hansen Solubility Parameters (HSPs) demonstrated good ability to extract carotenoid-rich PPFO, maintaining their relatively stable fatty acids composition and free acidity, showing that partial substitution of HC by ALC is technically possible.

Aprotic solvents effect on the UV-visible absorption spectra of bixin.

We describe here the effects of aprotic solvents on the spectroscopic characteristics of bixin. Bixin was dissolved in dimethyl sulfoxide, acetone, dichloromethane, ethyl acetate, chloroform, dimethyl carbonate, cyclohexane and hexane, separately, and its spectra in the resulting solutions were determined by UV-visible spectrophotometry at normal pressure and room temperature. We analyzed the effect of aprotic solvents on λmax according to Onsager cavity model and Hansen theory, and determined the approximate absorption coefficient with the Beer-Lambert law. We found that the UV-visible absorption spectra of bixin were found to be solvent dependent. The S0→S2 transition energy of bixin in solution was dependent principally on the refractive index of the solvents and the bixin-solvent dispersion interaction. There was a small influence of the solvents dielectric constant, permanent dipole interaction and hydrogen bonding occurred between bixin and solvents. The absorbance of bixin in various solvents, with the exception of hexane, increased linearly with concentration.

Alpha and omega of carotenoid cleavage.

In early 1900s, based on indirect evidence, Steenbock and Morton independently predicted that beta-carotene could be the biological precursor of vitamin A, although this notion was contested by others. In the 1930s, Thomas Moore showed the in vivo formation of vitamin A from beta-carotene. But it was not until Jim Olson and DeWitt Goodman independently showed in 1965 the formation of retinal, the aldehyde form of vitamin A from beta-carotene in cell-free extracts of liver and intestine, that this vital pathway of beta-carotene was recognized. Despite compelling evidence in several experimental systems for the central cleavage of beta-carotene to retinal by many investigators, there were some careful independent studies by Glover et al., Ganguly et al., Hansen and Meret and Krinsky et al. showing the eccentric cleavage of beta-carotene resulting in the formation of apocarotenoids both in vivo and in vitro. In an attempt to resolve this controversial issue, we revisited this problem in 1989 and showed beyond doubt the formation of retinal as the sole enzymatic product of a cytosolic enzyme from rabbit and rat intestinal mucosa by mass spectrometry and tracer analysis of the crystallized product. This was confirmed in 1996 by Nagao using the pig intestinal extract. Yeum et al. confirmed in 2000 that retinal is the sole product of beta-carotene cleavage in the presence of alpha-tocopherol, and that the observed formation of apocarotenoids occurs only in the absence of an antioxidant like alpha-tocopherol. In the same year, Barua and Olson also concluded from their in vivo studies in rats that central cleavage is by far the major pathway for the formation of vitamin A from beta-carotene. Beta, beta-carotene 15,15'-dioxygenase (EC 1.13.11.21) is the key enzyme that cleaves beta-carotene into two molecules of retinal. It is a cytosolic enzyme primarily localized in the duodenal mucosa although it has been found in liver. It is a 66 kDa sulfhydryl protein, requires molecular oxygen and is activated by ferrous ions. It is highly specific for 15:15' ethylenic bond of carotenoids although it has fairly broad specificity towards a number of carotenoids with at least one intact beta-ionone ring. The dioxygenase was recently cloned from Drosophila melanogaster and from the chicken intestine. The recombinant protein was found to form retinal as the sole cleavage product of beta-carotene. No apo-carotenoids were formed. Therefore, it is unequivocally proven that the major, if not the sole, pathway of beta-carotene cleavage to vitamin A is by oxidative cleavage of the central ethylenic bond of beta-carotene to yield two molecules of retinal. Most recently, human dioxygenase has also been cloned. Thus, the wisdom, vision and epoch-making mission of Jim Olson in the science of beta-carotene metabolism have been accomplished. I have no doubt that the impact of his original discovery of the dioxygenase and its importance in vitamin A nutriture should be forthcoming in the near future.

Murine leprosy and carotinoids

In this study of the effect of substances of the carotinoid type in the treatment of murine leprosy, it has been observed that there is a difference in the distribution of the Stefansky bacillus in the organism of mice according to whether the germs used in the inoculation are alive or have been killed by heat (autoclaving at 120ºC. for 20 minutes). Within the period of 80 days after inoculation, pieces of the liver were always negative when the inoculated germ was dead, and positive in the majority of cases when the germ was alive. Similar though less clear-cut findings were obtained with regard to the spleen. With infected animals treated with a crude substance of carotinoid type it was observed that the distribution of bacilli was the same as if they had been inoculated with dead germs, and that was also true when the treatment was with one of the fragments (fraction A) of the primitive substance. Two other fractions (b and C) were inactive, although it would appear probable that the active substance (fraction A) originates through an oxidation process from fraction C. A substance obtained in the laboratory through oxidation of raw carotinoid (fraction D) showed activity in a small number of observations.