Category Archives: Carotenoids

Bile salts: definition, functions, enterohepatic circulation, synthesis

Bile salts: contents in brief

What are bile salts

Bile salts and bile acids are polar cholesterol derivatives, and represent the major route for the elimination of the steroid from the body.
They are molecules with similar but not identical structures, and diverse physical and biological characteristics.
They are synthesized in the liver, stored in the gallbladder, secreted into the duodenum, and finally, for the most part, reabsorbed in the ileum.
Because at physiological pH these molecules are present as anions, the terms bile acid and bile salts are used herein as synonyms.

⇑ Back to the top ⇑

Chemical structure of bile salts

Bile Salts
Fig. 1 – Chemical Structures of the Most Abundant Bile Acids

Bile salts have similarities and differences with cholesterol molecule.
Like the steroid, they have a nucleus composed of four fused rings: three cyclohexane rings, labeled A, B and C, and a cyclopentane ring, labeled D. This structure is the perhydrocyclopentanophenanthrene, more commonly known as steroid nucleus.
In higher vertebrates, they have 24 carbon atoms, as the side chain is three carbons shorter than the original. In lower vertebrates, bile acids have 25, 26, or 27 carbon atoms. The side chain ends with a carboxyl group, ionized at pH 7, that can be linked to the amino acid glycine or taurine (see below).
In addition to the hydroxyl group at position 3, they have hydroxyl groups at positions 7 and/or 12.
All this makes them much more polar than cholesterol.

Bile Salts
Fig. 2 – Cholic Acid Structure

Since A and B rings are fused in cis configuration, the planar structure of the steroid nucleus is curved, and it is possible to identify:

  • a concave side, which is hydrophilic because the hydroxyl groups and the carboxyl group of the side chain, with or without the linked amino acid, are oriented towards it;
  • a convex side, which is hydrophobic because the methyl groups present at position 18 and 19 are orientated towards it.

Therefore, having both polar and nonpolar groups, they are amphiphilic molecules and excellent surfactants. However, their chemical structure makes them different from many other surfactants, often composed of a polar head region and a nonpolar tail.

⇑ Back to the top ⇑

Primary, conjugated and secondary bile salts

Bile Salts
Fig. 3 – Conjugated Bile Acids

Primary bile acids are those synthesized directly from cholesterol in the hepatocytes. In humans, the most important are cholic acid and chenodeoxycholic acid, which make up 80% of all bile acids. Before being secreted into the biliary tree, they are almost completely conjugated, up to 98%, with the glycine or taurine, to form glycoconjugates and tauroconjugates, respectively. In particular, approximately 75% of cholic acid and chenodeoxycholic acid are conjugated with glycine, to form glycocholic acid  and glycochenodeoxycholic acid, the remaining 25% with taurine, to form taurocholic acid and taurochenodeoxycholic.
Conjugated bile acids are molecules with more hydrophilic groups than unconjugated bile acids, therefore with a increased emulsifying capacity. In fact, conjugation decreases the pKa of bile acids, from about 6, a value typical of non-conjugated molecules, to about 4 for glycocholic acid, and about 2 for taurocholic acid. This makes that conjugated bile acids are ionized in a broader range of pH to form the corresponding salts.
The hydrophilicity of the common acid and bile salts decreases in the following order: glycine-conjugated < taurine-conjugated < lithocholic acid  < deoxycholic acid  < chenodeoxycholic acid < cholic acid <ursodeoxycholic acid.
Finally, conjugation also decreases the cytotoxicity of primary bile acids.

Secondary bile acids  are formed from primary bile acids which have not been reabsorbed from the small intestine. Once they reach the colon, they can undergo several modifications by colonic microbiota to form secondary bile acids (see below). They make up the remaining 20% of the body’s bile acid pool.

Another way of categorizing bile salts is based on their conjugation with glycine and taurine and their degree of hydroxylation. On this basis, three categories are identified.

  • Trihydroxy conjugates, such as taurocholic acid and glycocholic acid.
  • Dihydroxy conjugates, such as glycodeoxycholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, and taurodeoxycholic acid. They account for about 60% of bile salts present in the bile.
  • Unconjugated forms, such as cholic acid, deoxycholic acid, chenodeoxycholic acid, and lithocholic acid.

⇑ Back to the top ⇑

Function of bile acids

All their physiological functions are performed in the conjugated form.

  • They are the major route for the elimination of cholesterol from human body.
    Indeed, humans do not have the enzymes to break open the cyclohexane rings or  the cyclopentane ring of the steroid nucleus, nor to oxidize cholesterol to CO2 and water.
    The other mechanism to eliminate the steroid from the body is as cholesterol per se in the bile.
  • Bile salts are strong surfactants. And in particular, di- and trihydroxy conjugates are the best surfactants among bile acids, much more effective than unconjugated counterparts, since they have more polar groups.
    Once in contact with apolar lipids in the lumen of the small intestine, the convex apolar surface interacts with the apolar lipids, such as triglycerides, cholesterol esters, and ester of fat-soluble vitamins, whereas the concave polar surface interacts with the surrounding aqueous medium. This increases the dispersion of apolar lipids in the aqueous medium, as it allows the formation of tiny lipid droplets, increasing the surface area for:

lipase activity, mainly pancreatic lipase, (bile salts also play a direct role in the activation of this enzyme);

intestinal esterase activity.

Subsequently, they facilitate the absorption of lipid digestion products, as well as of fat soluble vitamins by the intestinal mucosa thanks to the formation of mixed micelles.
Bile acids perform a similar function in the gallbladder where, forming mixed micelles with phospholipids, they prevent the precipitation of cholesterol.
Note: as a consequence of the arrangement of polar and nonpolar groups, bile acids form micelles in aqueous solution, usually made up of less than 10 monomers, as long as their concentration is above the so-called critical micellar concentration or CMC.

  • At the intestinal level, they modulate the secretion of pancreatic enzymes and cholecystokinin.
  • In the small and large intestine, they have a potent antimicrobial activity, mainly deoxycholic acid, in particular against Gram-positive bacteria. This activity may be due to oxidative DNA damage, and/or to the damage of the cell membrane. Therefore, they play an important role in the prevention of bacterial overgrowth, but also in the regulation of gut microbiota composition.
  • In the last few years, it becomes apparent their regulatory role in the control of energy metabolism, and in particular for the hepatic glucose handling.

⇑ Back to the top ⇑

Enterohepatic circulation of bile salts

After fat intake, enteroendocrine cells of the duodenum secrete cholecystokinin into the blood stream. Hormone binding to receptors on smooth muscle cells of the gallbladder promotes their contraction; the hormone also causes the relaxation of the sphincter of Oddi. All this results in the secretion of the bile, and therefore of bile acids into the duodenum.
Under physiological conditions, human bile salt pool is constant, and equal to about 3-5 g. This is made possible by two processes:

  • their intestinal reabsorption;
  • their de novo synthesis (see below).

Up to 95% of the secreted bile salts is reabsorbed from the gut, not together with the products of lipid digestion, but through a process called enterohepatic circulation.
It is an extremely efficient recycling system, which seems to occur at least two times for each meal, and includes the liver, the biliary tree, the small intestine, the colon, and the portal circulation through which reabsorbed molecules return to the liver. Such recirculation is necessary since liver’s capacity to synthesize bile acids is limited and insufficient to satisfy intestinal needs if the bile salts were excreted in the feces in high amounts.
Most of the bile salts are reabsorbed into the distal ileum, the lower part of the small intestine, by a sodium-dependent transporter within the brush border of the enterocytes, called sodium-dependent bile acid transporter or ASBT, which carries out the cotransport of a molecule of bile acid and two sodium ions.
Within the enterocyte, it is thought that bile acids are transported across the cytosol to the basolateral membrane by the ileal bile acid-binding protein or IBABP. They cross the basolateral membrane by the organic solute transporter alpha-beta or OSTα/OSTβ, pass into the portal circulation, and, bound to albumin, reach the liver.
It should be noted that a small percentage of bile acids reach the liver through the hepatic artery.
A hepatic level, their extraction is very efficient, with a first-pass extraction fraction ranging from 50 to 90%, a percentage that depends on bile acid structure. The uptake of conjugated bile acids is mainly mediated by a Na+-dependent active transport system, that is, the sodium-dependent taurocholate cotransporting polypeptide or NTCP. However, a sodium-independent uptake can also occur, carried out by proteins of the family of organic anion transporting polypeptides or OATP, mainly OATP1B1 and OATP1B3.
The rate limiting step in the enterohepatic circulation is their canalicular secretion, largely mediated by the bile salt export pump or BSEP, in an ATP-dependent process. This pump carries monoanionic bile salts, which are the most abundant. Bile acids conjugated with glucuronic acid or sulfate, which are dianionic, are transported by different carriers, such as MRP2 and BCRP.

Note: serum levels of bile acids vary on the basis of the rate of their reabsorption, and therefore they are higher during meals, when the enterohepatic circulation is more active.

⇑ Back to the top ⇑

Intestinal metabolism of bile acids

Bile Salts
Fig. 4 – Intestinal Bile Acid Metabolism

Bile acids which escape ileal absorption pass into the colon where they partly undergo modifications by intestinal microbiota and are converted to secondary bile acids.
The main reactions are listed below.

  • Deconjugation
    On the side chain, hydrolysis of the C24 N-acyl amide bond can occur, with release of unconjugated bile acids and glycine or taurine. This reaction is catalyzed by bacterial hydrolases present both in the small intestine and in the colon.
  • 7α-Dehydroxylation
    Quantitatively, it is the most important reaction, carried out by colonic bacterial dehydratases that remove the hydroxyl group at position 7 to form 7-deoxy bile acids. In particular, deoxycholic acid is formed from cholic acid, and lithocholic acid, a toxic secondary bile acid, from chenodeoxycholic acid.
    It should be noted that 7α-dehydroxylation, unlike oxidation and epimerization (see below), can only occur on unconjugated bile acids, and therefore, deconjugation is an essential prerequisite.
  • Oxidation and epimerization
    They are reactions involving the hydroxyl groups at positions 3, 7 and 12, catalyzed by bacterial hydroxysteroid dehydrogenases. For example, ursodeoxycholic acid derives from the epimerization of chenodeoxycholic acid.

Some of the secondary bile acids are then reabsorbed from the colon and return to the liver. In the hepatocytes, they are reconjugated, if necessary, and resecreted. Those that are not reabsorbed, are excreted in the feces.
Whereas oxidations and deconjugations are carried out by a broad spectrum of anaerobic bacteria, 7α-dehydroxylations is carried out by a limited number of colonic anaerobes.
7α-Dehydroxylations and deconjugations increase the pKa of the bile acids, and therefore their hydrophobicity, allowing a certain degree of passive absorption across the colonic wall.
The increase of hydrophobicity is also associated with an increased toxicity of these molecules. And a high concentration of secondary bile acids in the bile, blood, and feces has been associated to the pathogenesis of colon cancer.

⇑ Back to the top ⇑

Soluble fibers and reabsorption of bile salts

The reabsorption of bile salts can be reduced by chelating action of soluble fibers, such as those found in fresh fruits, legumes, oats and oat bran, which bind them, decreasing their uptake. In turn, this increases bile acid de novo synthesis, up-regulating the expression of the 7α-hydroxylase and sterol 12α-hydroxylase (see below), and thereby reduces hepatocyte cholesterol concentration.
The depletion of hepatic cholesterol increases the expression of the LDL receptor, and thus reduces plasma concentration of LDL cholesterol. On the other hand, it also stimulates the synthesis of HMG-CoA reductase, the key enzyme in cholesterol biosynthesis.
Note: some anti-cholesterol drugs act by binding bile acids in the intestine, thereby preventing their reabsorption.

⇑ Back to the top ⇑

Synthesis of primary bile acids

Bile Salts
Fig. 5 – Primary Bile Acid Biosynthesis

Quantitatively, bile acids are the major product of cholesterol metabolism.
As previously said, enterohepatic circulation and their de novo synthesis maintain a constant bile acid pool size. In particular, de novo synthesis allows the replacement of bile salts excreted in the faces, about 5-10% of the body pool, namely ~ 0.5 g/day.
Below, the synthesis of cholic acid and chenodeoxycholic acid, and their conjugation with the amino acids taurine and glycine, is described.
There are two main pathways for bile acid synthesis: the classical pathway and the alternative pathway. In addition, some other minor pathways will also be described.

⇑ Back to the top ⇑

The classical or neutral pathway

In humans, up to 90% of bile salts are produced via the classical pathway (see fig. 5), also referred to as “neutral” pathway since intermediates are neutral molecules.
It is a metabolic pathway present only in the liver, that consists of reactions catalyzed by enzymes localized in the cytosol, endoplasmic reticulum, peroxisomes, and mitochondria, and whose end products are the conjugates of cholic acid and chenodeoxycholic acid.

  • The first reaction is the hydroxylation at position 7 of cholesterol, to form 7α-hydroxycholesterol. The reaction is catalyzed by cholesterol 7α-hydroxylase or CYP7A1 (E.C. 1.14.14.23). It is an enzyme localized in the endoplasmic reticulum, and catalyzes the rate-limiting step of the pathway.

Cholesterol + NADPH + H+ + O2 → 7α-Hydroxycholesterol + NADP+ + H2O

  • 7α-Hydroxycholesterol undergoes oxidation of the 3β-hydroxyl group and the shift of the double bond from the 5,6 position to the 4,5 position, to form 7α-hydroxy-4-cholesten-3-one. The reaction is catalyzed by 3β-hydroxy-Δ5-C27-steroid oxidoreductase or HSD3B7 (E.C. 1.1.1.181), an enzyme localized in the endoplasmic reticulum.
  • 7α-Hydroxy-4-cholesten-3-one can follow two routes:

to enter the pathway that leads to the synthesis of cholic acid, through the reaction catalyzed by 7α-hydroxy-4-cholesten-3-one 12α-monooxygenase or sterol 12α-hydroxylase or CYP8B1 (E.C. 1.14.18.8), an enzyme localized in the endoplasmic reticulum;

to enter the pathway that leads the synthesis of chenodeoxycholic acid, through the reaction catalyzed by 3-oxo-Δ4-steroid 5β-reductase or AKR1D1 (E.C. 1.3.1.3), a cytosolic enzyme.

It should be underlined that the activity of sterol 12α-hydroxylase determines the ratio of cholic acid to chenodeoxycholic acid, and, ultimately, the detergent capacity of bile acid pool. And in fact, the regulation of sterol 12α-hydroxylase gene transcription is one of the main regulatory step of the classical pathway.

Therefore, if 7α-hydroxy-4-cholesten-3-one proceeds via the reaction catalyzed by sterol 12α-hydroxylase, the following reactions will occur.

  • 7α-Hydroxy-4-cholesten-3-one is hydroxylated at position 12 by sterol 12α-hydroxylase, to form 7α,12α-dihydroxy-4-cholesten-3-one.
  • 7α,12α-Dihydroxy-4-cholesten-3-one undergoes reduction of the double bond at 4,5 position, in the reaction catalyzed by 3-oxo-Δ4-steroid 5β-reductase, to form 5β-cholestan-7α,12α-diol-3-one.
  • 5β-Cholestan-7α,12α-diol-3-one undergoes reduction of the hydroxyl group at position 4, in the reaction catalyzed by 3α-hydroxysteroid dehydrogenase or AKR1C4 (EC 1.1.1.213), a cytosolic enzyme, to form 5β-cholestan-3α,7α,12α-triol.
  • 5β-Cholestan-3α,7α,12α-triol undergoes oxidation of the side chain via three reactions catalyzed by sterol 27-hydroxylase or CYP27A1 (EC 1.14.15.15). It is a mitochondrial enzyme also present in extrahepatic tissues and macrophages, which introduces a hydroxyl group at position 27. The hydroxyl group is oxidized to aldehyde, and then to carboxylic acid, to form 3α,7α,12α-trihydroxy-5β-cholestanoic acid.
  • 3α,7α,12α-Trihydroxy-5β-cholestanoic  acid is activated to its coenzyme A ester, 3α,7α,12α-trihydroxy-5β-cholestanoyl-CoA, in the reaction catalyzed by either very long chain acyl-CoA synthetase or VLCS (EC 6.2.1.-), or bile acid CoA synthetase or BACS (EC 6.2.1.7), both localized in the endoplasmic reticulum.
  • 3α,7α,12α-Trihydroxy-5β-cholestanoyl-CoA is transported to peroxisomes where it undergoes five successive reactions, each catalyzed by a different enzyme. In the last two reactions, the side chain is shortened to four carbon atoms, and finally cholylCoA is formed.
  • In the last step, the conjugation, via amide bond, of the carboxylic acid group of the side chain with the amino acid glycine or taurine occurs. The reaction is catalyzed by bile acid-CoA:amino acid N-acyltransferase or the BAAT (EC 2.3.1.65), which is predominantly localized in peroxisomes.
    The reaction products are thus the conjugated bile acids: glycocholic acid and taurocholic acid.

If 7α-hydroxy-4-cholesten-3-one does not proceed via the reaction catalyzed by sterol 12α-hydroxylase, it enters the pathway that leads to the synthesis of chenodeoxycholic acid conjugates, through the reactions described below.

  • 7α-Hydroxy-4-cholesten-3-one is converted to 7α-hydroxy-5β-cholestan-3-one in the reaction catalyzed by 3-oxo-Δ4-steroid 5β-reductase.
  • 7α-Hydroxy-5β-cholestan-3-one is converted to 5β-cholestan-3α,7α-diol in the reaction catalyzed by 3α-hydroxysteroid dehydrogenase.

Then, the conjugated bile acids glycochenodeoxycholic acid and taurochenodeoxycholic acid are formed by modifications similar to those seen for the conjugation of cholic acid, and catalyzed mostly by the same enzymes.

Note: unconjugated bile acids formed in the intestine must reach the liver to be reconjugated.

⇑ Back to the top ⇑

The alternative or acidic pathway

It is prevalent in the fetus and neonate, whereas in adults it leads to the synthesis of less than 10% of the bile salts.
This pathway  (see fig. 5) differs from the classical pathway in that:

  • the intermediate products are acidic molecules, from which the alternative name “acidic pathway”;
  • the oxidation of the side chain is followed by modifications of the steroid nucleus, and not vice versa;
  • the final products are conjugates of chenodeoxycholic acid.

The first step involves the conversion of cholesterol into 27-hydroxycholesterol in the reaction catalyzed by sterol 27-hydroxylase.
27-Hydroxycholesterol can follow two routes.

Route A

  • 27-hydroxycholesterol is converted to 3β-hydroxy-5-cholestenoic acid in a reaction catalyzed by sterol 27-hydroxylase.
  • 3β-Hydroxy-5-cholestenoic acid is hydroxylated at position 7 in the reaction catalyzed by oxysterol 7α-hydroxylase or CYP7B1 (EC 1.14.13.100), an enzyme localized in the endoplasmic reticulum, to form 3β-7α-dihydroxy-5-colestenoic acid.
  • 3β-7α-Dihydroxy-5-cholestenoic acid is converted to 3-oxo-7α-hydroxy-4-cholestenoic acid, in the reaction catalyzed by 3β-hydroxy-Δ5-C27-steroid oxidoreductase.
  • 3-Oxo-7α-hydroxy-4-cholestenoic acid, as a result of side chain modifications, forms chenodeoxycholic acid, and then its conjugates.

Route B

  • 27-Hydroxycholesterol is converted to 7α,27-dihydroxycholesterol in the reaction catalyzed by oxysterol 7α-hydroxylase and cholesterol 7α-hydroxylase.
  • 7α,27-Dihydroxycholesterol is converted to 7α,26-dihydroxy-4-cholesten-3-one in the reaction catalyzed by 3β-hydroxy-Δ5-C27-steroid oxidoreductase;

7α, 26-Dihydroxy-4-cholesten-3-one can be transformed directly to conjugates of chenodeoxycholic acid, or can be converted to 3-oxo-7α-hydroxy-4-colestenoic acid,  and then undergo side chain modifications and other reactions that lead to the synthesis of the conjugates of chenodeoxycholic acid.

⇑ Back to the top ⇑

Minor pathways

There are also minor pathways (see fig. 5) that contribute to bile salt synthesis, although to a lesser extent than classical and alternative pathways.

For example:

  • A cholesterol 25-hydroxylase (EC 1.14.99.38) is expressed in the liver.
  • A cholesterol 24-hydroxylase or CYP46A1 (EC 1.14.14.25) is expressed in the brain, and therefore, although the organ cannot export cholesterol, it exports oxysterols.
  • A nonspecific 7α-hydroxylase has also been discovered. It is  expressed in all tissues and appears to be involved in the generation of oxysterols, which may be transported to hepatocytes to be converted to chenodeoxycholic acid.

Additionally, sterol 27-hydroxylase is expressed in various tissues, and therefore its reaction products must be transported to the liver to be converted to bile salts.

⇑ Back to the top ⇑

Bile salts: regulation of synthesis

Regulation of bile acid synthesis occurs via a negative feedback mechanism, particularly on the expression of cholesterol 7α-hydroxylase and sterol 12α-hydroxylase.
When an excess of bile acids, both free and conjugated, occurs, these molecules bind to the nuclear receptor farnesoid X receptor or FRX, activating it: the most efficacious bile acid is chenodeoxycholic acid, while others, such as ursodeoxycholic acid, do not activate it.
FRX induces the expression of the transcriptional repressor small heterodimer partner or SHP, which in turn interacts with other transcription factors, such as liver receptor homolog-1 or LRH-1, and hepatocyte nuclear factor-4α or HNF-4α. These transcription factors bind to a sequence in the promoter region of 7α-hydroxylase and 12α-hydroxylase genes, region called bile acid response elements or BAREs, inhibiting their transcription.
One of the reasons why bile salt synthesis is tightly regulated is because many of their metabolites are toxic.

⇑ Back to the top ⇑

References

Chiang J.Y.L. Bile acids: regulation of synthesis. J Lipid Res 2009;50(10):1955-66. doi:10.1194/jlr.R900010-JLR200

Gropper S.S., Smith J.L. Advanced nutrition and human metabolism. 6h Edition. Cengage Learning, 2012 [Google eBook]

Moghimipour E., Ameri A., and Handali S. Absorption-enhancing effects of bile salts. Molecules 2015;20(8); 14451-73. doi:10.3390/molecules200814451

Monte M.J., Marin J.J.G., Antelo A., Vazquez-Tato J. Bile acids: Chemistry, physiology, and pathophysiology. World J Gastroenterol 2009;15(7):804-16. doi:10.3748/wjg.15.804

Rawn J.D. Biochimica. Mc Graw-Hill, Neil Patterson Publishers, 1990

Rosenthal M.D., Glew R.H. Medical biochemistry – Human metabolism in health and disease. John Wiley J. & Sons, Inc., Publication, 2009

Sundaram S.S., Bove K.E., Lovell M.A. and Sokol R.J. Mechanisms of Disease: inborn errors of bile acid synthesis. Nat Clin Pract Gastroenterol Hepatol 2008;5(8):456-68. doi:10.1038/ncpgasthep1179

Chemical composition of olive oil

Chemical composition of olive oil: contents in brief

Olive oil constituents

Olive Oil
Fig. 1 – EVOO

From a chemical point of view, we can identify in the olive oil two fractions, depending on the behavior in the presence of heating and strong alkaline solutions (concentrated solutions of KOH or NaOH):

  • the saponifiable fraction, which represents 98-99% of the total weight, is composed of substances that form soaps in the above conditions;
  • the unsaponifiable fraction, which represents the remaining 1-2% of the total weight, is composed of substances that fail to form soaps in the above conditions.

⇑ Back to the top ⇑

Saponifiable fraction of olive oil

It is composed of saturated and unsaturated fatty acids, esterified almost entirely to glycerol to form triglycerides (or triacylglycerols). To a much lesser extent, diglycerides (or diacylglycerols), monoglycerides (monoacylglycerols), and free fatty acids are also found.
Unsaturated fatty acids make up 75 to 85% of the total fatty acids. Oleic (O) and linoleic (L) acids are the most abundant ones; palmitoleic, eptadecenoic, gadoleic and alpha-linolenic (Ln) acids are present in lower/trace amounts.

Oleic Acid
Fig. 2 – IOOC and Fatty Acids

Oleic acid is the major fatty acid in olive oils. According to the rules laid down by the International Olive Oil Council (IOOC), its concentration must range from 55% to 83% of total fatty acids.
Linoleic acid is the most abundant polyunsaturated fatty acid in olive oil; its concentration must vary between 2.5% and 21% (IOOC). Because of its high degree of unsaturation, it is subject to oxidation; this means that an oil high in linoleic acid becomes rancid easily, and thus it may be stored for a shorter time.
In a Mediterranean-type diet, olive oil is the main source of fat: therefore, oleic acid, among monounsaturated fatty acids, and linoleic acid, among polyunsaturated fatty acids, are the most abundant fatty acids.
alpha-Linolenic acid must be present in very low amount, according to the IOOC standards ≤1%. It is an omega-3 polyunsaturated fatty acid, which may have health benefits. However, because of to its high degree of unsaturation (higher than that of linoleic acid), it is very susceptible to oxidation, and therefore it promotes rancidity of the olive oil that contains it.
Saturated fatty acids make up 15 to 25% of the total fatty acids.
Palmitic (P) (7.5-20%) and stearic (S) acids (0.5-5%) are the most abundant saturated fatty acids; myristic, heptadecanoic, arachidic, behenic and lignoceric acids may be present in trace amounts.

The presence of fatty acids that should be absent or present in amounts different than those found is a marker of adulteration with other vegetable oils. On this regard, particular attention is paid to myristic, arachidic, behenic, lignoceric, gadoleic and alpha-linolenic acids, whose limits are set by IOOC.

Fatty acid composition is influenced by several factors.

  • The climate.
  • The latitude.
  • The zone of production.
    Italian, Spanish and Greek olive oils are high in oleic acid and low in palmitic and linoleic acids, while Tunisian olive oils are high in palmitic and linoleic acids but lower in oleic acid. Therefore, oils can be divided into two groups:

one rich in oleic acid and low in palmitic and linoleic acids;
the other high in palmitic and linoleic acids and low in oleic acid.

  • The cultivar.
  • The degree of olive ripeness at the time of oil extraction.
    It should be noted that oleic acid is formed first in the fruit, and data seem to indicate a competitive relationship between oleic acid and palmitic, palmitoleic, and linoleic acids.

⇑ Back to the top ⇑

Triglycerides of olive oil

Olive Oil
Fig. 3 – The sn Positions of Triglycerides

As previously said, fatty acids in olive oil are almost entirely present as triglycerides.
In small percentage, they are also present as diglycerides, monoglycerides, and in free form.
During triglyceride biosynthesis, thanks to the presence of specific enzymes, only about 2% of glycerol binds palmitic acid in the sn-2 position (also the percentage of stearic acid in the sn-2 position is very low); for the most part, the sn-2 position is occupied by oleic acid.
On the contrary, if we consider oils that have undergone a nonenzymatic esterification, the percentage of palmitic acid in the sn-2 position increases significantly.
Note: sn = stereospecific numbering

Among triglycerides present in significant proportions in olive oil, there are:

  • OOO: 40-59%;
  • POO: 12-20%;
  • OOL: 12.5-20%;
  • POL:  5.5-7%;
  • SOO: 3- 7%.

POP, POS, OLnL, OLnO, PLL, PLnO are present in smaller amounts.
Trilinolein (LLL) is a triglyceride that contains three molecules of linoleic acid. Its low content is an indicator of an oil of good quality.
Triglycerides containing three saturated fatty acids or three molecules of alpha-linolenic acid have not been reported.

⇑ Back to the top ⇑

Diglycerides and monoglycerides of olive oil

Their presence is due to an incomplete synthesis and/or a partial hydrolysis of triglycerides.
The content of diglycerides in virgin olive oil ranges from 1% to 2.8%. 1,2-Diglycerides prevail in fresh olive oil, representing over 80% of the diglycerides. During oil storage, isomerization occurs with a progressive increase of the more stable 1-3 isomers, which after about 10 months become the major isomers.
Therefore, the ratio 1,2/1,3-diglycerides may be used as an indicator of the age of the oil.
Monoglycerides are present in amounts lower than diglycerides, <0.25%, with 1-monoglycerides far more abundant than 2-monoglycerides.

⇑ Back to the top ⇑

Unsaponifiable fractions of olive oil

It is composed of a large number of different molecules, very important from a nutritional point of view, as they contribute significantly to the health effects of olive oil.
Furthermore, they are responsible for the stability and the taste of olive oil, and are also used to detect adulteration with other vegetable oils.
This fraction includes tocopherols, sterols, polyphenols, pigments, hydrocarbons, aromatic and aliphatic alcohol, triterpene acids, waxes, and minor constituents.
Their content is influenced by factors similar to those seen for fatty acid composition, such as:

  • the cultivar;
  • the degree of ripeness of the olive;
  • the zone of production;
  • the crop year and olive harvesting practices;
  • the storage time of olives;
  • the oil extraction process;
  • the storage conditions of the oil.

It should be noted that many of these compounds are not present in refined olive oils, as they are removed during the refining processes.

⇑ Back to the top ⇑

Polyphenols

They make up 18 to 37% of the unsaponifiable fraction.
They are a very heterogeneous group of molecules with nutritional and organoleptic properties  (for example, oleuropein and hydroxytyrosol give oil its bitter and pungent taste).
For a more extensive discussion, see: ” Polyphenols in olive oil: variability and composition.”

⇑ Back to the top ⇑

Hydrocarbons

Olive Oil
Fig. 4 – Squalene

They make up 30 to 50% of the unsaponifiable fraction.
Squalene and beta-carotene are the main molecules.
Squalene, isolated for the first time from shark liver, is the major constituent of the unsaponifiable fraction, and constitutes more than 90% of the hydrocarbons. Its concentration ranges from 200 to 7500 mg/kg of olive oil.
It is an intermediate in the biosynthesis of the four-ring structure of steroids, and it seems to be responsible of several health effects of olive oil.
In the hydrocarbon fraction of virgin olive oil, n-paraffins, diterpene and triterpene hydrocarbons, isoprenoidal polyolefins are also found.
Beta-carotene acts both as antioxidant, protecting oil during storage, and as dye (see below).

⇑ Back to the top ⇑

Sterols

They are important lipids of olive oil, and are:

  • linked to many health benefits for consumers;
  • important to the quality of the oil;
  • widely used for checking its genuineness.
    On this regard, it is to underline that sterols are species-specific molecules; for example, the presence of high concentrations of brassicasterol, a sterol typically found in Brassicaceae (Cruciferae) family, such as rapeseed, indicates adulteration of olive oil with canola oil.

Four classes of sterols are present in olive oil: common sterols, 4-methylsterols, triterpene alcohols, and triterpene dialcohols. Their content ranges from 1000 mg/kg, the minimum value required by the IOOC standard, to 2000 mg/kg. The lowest values are found in refined oils because of the refining processes may cause losses up to 25%.

⇑ Back to the top ⇑

Common sterols or 4α-desmethylsterols
Olive Oil
Fig. 5 – beta-Sitosterol

Common sterols are present mainly in the free and esterified form; however they have been also found as lipoproteins and sterylglucosides.
The main molecules are beta-sitosterol, which makes up 75 to 90% of the total sterol, Δ5-avenasterol, 5 to  20%, and campesterol, 4%. Other components found in lower amounts or traces are, for example, stigmasterol, 2%, cholesterol, brassicasterol, and ergosterol.

⇑ Back to the top ⇑

4-Methylsterols

They are intermediates in the biosynthesis of sterols, and are present both in the free and esterified form. They are present in small amounts, much lower than those of common sterols and triterpene alcohols, varying between 50 and 360 mg/kg. The main molecules are obtusifoliol, cycloeucalenol, citrostadienol, and gramisterol.

⇑ Back to the top ⇑

Triterpene alcohols or 4,4-dimethylsterols

They are a complex class of sterols, present both in the free and esterified form. They are found in amounts ranging from 350 to 1500 mg/kg.
The main components are beta-amyrin, 24-methylenecycloartanol, cycloartenol, and butyrospermol; other molecules present in lower/trace amounts are, for example, cyclosadol, cyclobranol, germanicol, and dammaradienol.

⇑ Back to the top ⇑

Triterpene dialcohols

The main triterpene dialcohols found in olive oil are erythrodiol and uvaol.
Erythrodiol is present both in the free and esterified form; in virgin olive oil, its level varies between 19 and 69 mg/kg, and the free form is generally lower than 50 mg/kg.

⇑ Back to the top ⇑

Tocopherols

They make up 2 to 3% of the unsaponifiable fraction, and include vitamin E.
Of the eight E-vitamers, alpha-tocopherol represents about 90% of tocopherols in virgin olive oil. It is present in the free form and in very variable amount, but on average higher than 100 mg/kg of olive oil. Thanks to its in vivo antioxidant properties, its presence is a protective factor for health. Alpha-tocopherol concentration seems to be related to the high levels of chlorophylls and to the concomitant requirement for deactivation of singlet oxygen.
Beta-tocopherol, delta-tocopherol, and gamma-tocopherol are usually present in low amounts.

⇑ Back to the top ⇑

Pigments

In this group we find chlorophylls and carotenoids.
In olive oil, chlorophylls are present as phaeophytins, mainly  phaeophytin a (i.e. a chlorophyll from which magnesium has been removed and substituted with two hydrogen ions), and confer the characteristic green color to olive oil. They are photosensitizer molecules that contribute to the photooxidation of olive oil itself.
Beta-carotene and lutein are the main carotenoids in olive oil. Several xanthophylls are also present, such as antheraxanthin, beta-cryptoxanthin, luteoxanthin, mutatoxanthin, neoxanthin, and violaxanthin.
Olive oil’s color is the result of the presence of chlorophylls and carotenoids and of their green and yellow hues. Their presence is closely related.

⇑ Back to the top ⇑

Triterpene acids

They are important components of the olive, and are present in trace amounts in the oil.
Oleanolic and maslinic acids are the main triterpene acids in virgin olive oil: they are present in the olive husk, from which they are extracted in small amount during processing.

⇑ Back to the top ⇑

Aliphatic and aromatic alcohols

Fatty alcohols and diterpene alcohols are the most important ones.
Aliphatic alcohols have a number of carbon atoms between 20 and 30, and are located mostly inside the olive stones, from where they are partially extracted by milling.

⇑ Back to the top ⇑

Fatty alcohols

They are linear saturated alcohols with more than 16 carbon atoms.
They are found in the free and esterified form and are present, in virgin olive oil, in amount not generally higher than 250 mg/kg.
Docosanol (C22), tetracosanol (C24), hexacosanol (C26), and octacosanol (C28) are the main fatty alcohols in olive oil, with tetracosanol and hexacosanol present in larger amounts.
Waxes, which are minor constituents of olive oil, are esters of fatty alcohols with fatty acids, mainly of palmitic acid and oleic acid. They can be used as a criterion to discriminate between different types of oils; for example, they must be present in virgin and extra virgin olive oil at levels <150 mg/kg, according to the IOOC standards.

⇑ Back to the top ⇑

Diterpene alcohols

Geranylgeraniol and phytol are two acyclic diterpene alcohols, present in the free and esterified form. Among esters present in the wax fraction of extra virgin olive oil, oleate, eicosenoate , eicosanoate, docosanoate, and tetracosanoate have been found, mainly as phytyl derivatives.

⇑ Back to the top ⇑

Volatile compounds

More than 280 volatile compounds have been identified in olive oil, such as hydrocarbons, the most abundant fraction, alcohols, aldehydes, ketones, esters, acids, ethers and many others. However, only about 70 of them are present at levels higher than the perception threshold beyond which they may contribute to the aroma of virgin olive oil.

⇑ Back to the top ⇑

Minor components

Phospholipids are found among the minor components of olive oil; the main ones are phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol.
In the unfiltered oils, trace amounts of proteins may be found.

⇑ Back to the top ⇑

References

Gunstone F.D. Vegetable oils in food technology: composition, properties and uses. 2th Edition. Wiley J. & Sons, Inc., Publication, 2011

Pasqualone A., Sikorska E., Gomes T. Influence of the exposure to light on extra virgin olive oil quality during storage. Eur Food Res Technol 2005;221:92-8. doi:10.1007/s00217-004-1126-8

Servili M., Sordini B., Esposto S., Urbani S., Veneziani G., Di Maio I., Selvaggini R. and Taticchi A. Biological activities of phenolic compounds of extra virgin olive oil. Antioxidants 2014;3:1-23. doi:10.3390/antiox3010001

Human health and carotenoids

Benefits of carotenoids for human health

Carotenoids belong to the category of bioactive compounds taken up with diet, that is, molecules able to provide protection against many diseases such as cardiovascular diseases, cancer and macular degeneration. They are also important for the proper functioning of the immune system.
Among the mechanisms that seem to be at the basis of their human health-promoting effects have been reported (Olson, 1999, see References):

  • the capability to quench singlet oxygen (see above);
  • the scavenging of peroxyl radicals and reactive nitrogen species;
  • the modulation of carcinogen metabolism;
  • the inhibition of cell proliferation;
  • the enhancement of the immune response;
  • a filtering action of blue light;
  • the enhancement of cell differentiation;
  • stimulation of cell-to-cell communication

Carotenoids, antioxidant activity and human health

Human Health and Carotenoids
Fig. 1 – Free Radical

Carotenoids, with the adaptation of organisms to aerobic environment, and therefore to the presence of oxygen, have offered protection against oxidative damage from free radicals, particularly by singlet oxygen, a powerful oxidizing agent (see also below).
Carotenoids stabilize singlet oxygen acting both chemical and physical point of view:

  • chemical action involves the union between the two molecules;
  • in physical action, the radical transfers its excitation energy to the carotenoid. The result is a low energy free radical and an excited carotenoid; later, the energy acquired by the carotenoid is released as heat to the environment, and the molecule, that remains intact, is ready to carry out another cycle of stabilization of singlet oxygen, and so on.

The capability of carotenoids to quench singlet oxygen is due to the conjugated double-bond system present in the molecule, and the maximum protection is given by those molecules that have nine or more double bonds (moreover, the presence of oxygen in the molecule, as in xanthophylls, seems to have a role).
Carotenoids are involved not only in singlet oxygen quenching, but also in the scavenging of other reactive species both of oxygen, as peroxyl radicals (therefore contributing to the reduction of lipid peroxidation) and nitrogen. These reactive molecules are generated during the aerobic metabolism but also in the pathological processes.

Lycopene, xanthophylls and human health

Lycopene, a carotene, canthaxanthin and astaxanthin, two xanthophylls present in foods of animal origin, are better antioxidants than beta-carotene but also than zeaxanthin that, with lutein, is involved in prevention of age-related macular degeneration.
Lycopene, in addition to act on oxygen free radicals, acts as antioxidant also on the radicals of vitamin C and vitamin E, that are generated during the antioxidant processes in which these vitamins are involved, “repairing them”.
Finally, lycopene exerts its antioxidant action also indirectly, inducing the synthesis of enzymes involved in the protection against the action of oxygen free radicals and other electrophilic species; these enzymes are quinone reductase, glutathione S-transferase and superoxide dismutase (they are part of the enzymatic antioxidant system).

Vitamin A and human health

Human Health and Vitamin A
Fig. 2 – Provitamin A Activity

Vitamin A, whose deficiency affects annually more than 100 million children worldwide, causing more than a million deaths and half million cases of blindness, is a well-known carotenoid derivative with many biological actions, being essential for reproduction, growth, vision, immune function and general human health.
In the human diet, the major sources of vitamin A are the preformed vitamin, which is found in foods of animal origins (meat, milk, eggs, etc), and provitamin A carotenoids, present in fruits and vegetables. In economically deprived countries, fruits and vegetables are the main source of vitamin A being less expensive than food of animal origin.
Of the more than 750 different carotenoids identified in natural sources, only about 50 have provitamin A activity, and among these, beta-carotene (precisely, all-trans-beta-carotene isomer) is the main precursor of the vitamin A.
Among the other carotenoids precursors of vitamin A, alpha-carotene, gamma-carotene, beta-cryptoxanthin, alpha-cryptoxanthin, and beta-carotene-5,6-epoxide have about half the bioactivity of beta-carotene.
Spinach, carrots, pumpkins, sweet potatoes (yellow) are example of vegetables rich in beta-carotene and other provitamin A carotenoids.
Acyclic carotenes, such as lycopene (the main carotenoid in the human diet), and xanthophylls, except those mentioned above (beta-cryptoxanthin, alpha-cryptoxanthin, and beta-carotene-5,6-epoxide), cannot be converted to vitamin A.

References

de la Rosa L.A., Alvarez-Parrilla E., Gonzàlez-Aguilar G.A. Fruit and vegetable phytochemicals: chemistry, nutritional value, and stability. 1th Edition. Wiley J. & Sons, Inc., Publication, 2010

Johnson E.J. The role of carotenoids in human health. Nutr Clin Care 2002;5(2):56-65 [Abstract]

Olson, J.A. 1999. Carotenoids. p. 525-541. In: Shils M.E., Olson J.A., Shike M., Ross A.C. “Modern nutrition in health and disease” 9th ed., by Lippincott, Williams & Wilkins, 1999

Ross A.B., Thuy Vuong L., Ruckle J., Synal H.A., Schulze-König T., Wertz K., Rümbeli R., Liberman R.G., Skipper P.L., Tannenbaum S.R., Bourgeois A., Guy P.A., Enslen M., Nielsen I.L.F., Kochhar S., Richelle M., Fay L.B., and Williamson G. Lycopene bioavailability and metabolism in humans: an accelerator mass spectrometry study. Am J Clin Nutr 2011;93:1263-73 [Abstract]

 

Functions of carotenoids in plants and foods

Through the course of evolution, carotenoids, thank to their unique physical and chemical properties, have proven to be highly versatile molecules, being able to perform many functions in many different organisms, like plants.

Carotenoids in photosynthesis

Carotenoids, in the early stages of the emergence of single-celled photosynthetic organisms, are probably been used for light harvesting at wavelengths different from those covered by chlorophyll. Therefore carotenoids, acting as light absorbing accessory pigments, have allowed to expand the range of solar radiation absorbed and so utilized for photosynthesis, energy that is then transferred to chlorophyll itself.
The major carotenoids involved in light harvesting, that accumulate in green plant tissues, are beta-carotene, lutein, neoxanthin, and violaxanthin, that absorb light energy in the 400- to 500-nm range.
Moreover, they protect chlorophyll from photooxidation (in humans, they may contribute to the protection of photo-oxidative damage caused by UV rays, thus acting as a endogenous photo-protective agents).

Carotenoids and autumn leaf color

Carotenoids and Plants: Autumn Leaf Color
Fig. 1 – Carotenoids and Autumn Leaf Color

Leaf color of deciduous plants in different seasons, green, yellow, orange or red, is due to the presence in them of natural pigments.
In spring and summer, the predominant pigment present in the leaf is chlorophyll, and therefore the color is green.
During the fall, the color changes from green to yellow, orange or red, depending on the type of plant: this is a consequence of the change, both qualitative and quantitative, in the pigment content.
In fact, as a result of the decrease of the temperature and daylight hours, the production of chlorophyll is interrupted and that already present is demolished into colorless metabolites. In this way the predominant pigments become carotenoids (yellow-orange), molecules much more stable than the chlorophyll, which remain in the leaf coloring it (it do not seem to be synthesized de novo), and anthocyanins (red-purple), which, unlike carotenoids, are not present during the growing season, but are synthesized in autumn, just before leaf fall. Therefore, it can be concluded that the red-purple color assumed from the leaves of certain plants is not a side effect of leaf senescence but results from anthocyanins de-novo synthesis.
Depending on the prevalence of carotenoids or anthocyanins, leaf color changes from green to yellow/orange, as in Ginkgo biloba (yellow), or red-purple as in some maples.

And plants with non green leaves?
Their color is not due to the absence of chlorophyll but the presence of very high amounts of other pigments, typically carotenoids and anthocyanins, that “cover” the chlorophyll, determining the color of the leaf.

Some functions of apocarotenoids in plants and foods

These oxygenated carotenoids, containing fewer than 40 carbon atoms, have many functions in plants and animals and are also important for the aroma and flavor of foods.
Some of their main functions include the following.

  • Apocarotenoids have significant roles in the response signals involved in the development and in the response to the environment (for example abscisic acid).
  • They can act as visual or volatile signals to attract pollinators.
  • They are important in the defense mechanisms of plants.
  • They have a role in regulating plant architecture.
  • An apocarotenal, trans-beta-apo-8′-carotenal, found in citrus fruits and spinach, with a low provitamin A activity, is used in pharmaceuticals and cosmetics, and is also a food additive (E160e) legalized by the European Commission for human consumption.
  • Apocarotenoids make an important contribution to the nutritional quality and flavor of many types of foods such as fruits, wine and tea. Two natural apocarotenoids, crocetin and bixina, have economic importance as they are used as pigments and aroma in foods.
  • Finally, a broad range of apocarotenals derive from oxidative reactions that occur in food processing; these molecules are intermediates in the formation of smaller molecules, important for the color and flavor of the food.
References

Archetti, M., Döring T.F., Hagen S.B., Hughes N.M., Leather S.R., Lee D.W., Lev-Yadun S., Manetas Y., Ougham H.J. Unravelling the evolution of autumn colours: an interdisciplinary approach. Trends Ecol Evol 2009;24(3):166-73 [Abstract]

de la Rosa L.A., Alvarez-Parrilla E., Gonzàlez-Aguilar G.A. Fruit and vegetable phytochemicals: chemistry, nutritional value, and stability. 1th Edition. Wiley J. & Sons, Inc., Publication, 2010

Carotenoids: definition, structure and classification

What are carotenoids?

A Good Source of Carotenoids: Carrots
Fig. 1 – Carrots

Carotenoids are soluble-fat pigments found throughout nature.
Carotenoids were discovered during the 19th century.

  • In 1831 Wachen proposed the term “carotene” for a pigment crystallized from carrot roots;
  • Berzelius called the more polar yellow pigments extracted from autumn leaves “xanthophylls” (originally phylloxanthins), from Greek words xanthos, meaning yellow, and phyllon, meaning leaf;
  • Tswett separated many pigments and called them “carotenoids.”

They occur in the chromoplasts of plants and some other photosynthetic organisms such as algae and in some types of fungi and bacteria; they are also produced by some invertebrates (Aphids).
There are more than 750 different carotenoids ranging in color from red (such as lycopene), to orange (such as alpha-carotene, beta-carotene, and gamma-carotene) or yellow (such as lutein, alfa-cryptoxanthin or violaxanthin); more than 100 have been found in fruits and vegetables.
In some green plants and in their parts, generally the darker the green color, the higher the carotenoid content: for example, carotenoid content in pale green cabbage is less than 1% of that in dark green one.
Fruit carotenoids are very different, and those present in ripe fruits may be different from those present in unripe fruits.
They also occur extensively in microorganisms and animals.
In plants, microorganism and animals carotenoids have diverse and important functions and actions.

Chemical structure of carotenoids

Carotenoids are a class of hydrocarbon compounds consisting of 40 carbon atoms (tetraterpenes), with a structure characterized by an extensive conjugated double-bond system that determines the color (it serves as a light-absorbing chromophore): as the number of conjugated double-bond increases, color changes from pale yellow, to orange, to red.
In nature, they exist primarily in the more stable all-trans isomeric configuration, even though small amounts of cis isomers do occur too (they can be produced from all-trans forms also during processing).
Traditionally, carotenoids have been given trivial names derived from the biological source from which they are extracted. However, a semisystematic scheme exists: it allows carotenoids to be named in a way that describes and defines their structure.

Classification of carotenoids

Depending on the presence or absence of oxygen in the molecule, they can be divided into:

  • xanthophylls, which contain oxygen, such as:

Antheraxanthin
Astaxanthin (red)
Auroxanthin
Bixin, E160b
Canthaxanthin (red), E161g
Capsanthin, E160c
Capsorubin, E160c
beta-Carotene-5,6-epoxide
alfa-Cryptoxanthin (yellow)
beta-Cryptoxanthin (orange)
Crocetin
Lutein (yellow), E161b
Lutein-5,6-epoxide or taraxanthin
Luteoxanthin
Lycophyll
Lycoxanthin
Neoxanthin
Rubixanthin
Tunaxanthin
Violaxanthin (yellow)
Zeaxanthin (yellow-orange)
Zeinoxanthin

  • carotenes, which lack oxygen, as such:

alfa-Carotene (orange)
beta-Carotene (orange), E160a
delta-Carotene
gamma-Carotene (orange)
Lycopene (red), E160d
Neurosporene
Phytoene (colorless)
Phytofluene
alfa-Zeacarotene
beta-Zeacarotene
zeta-Carotene

Depending on chemical structure they can be divided into:

  • acyclic carotenes: formed by a linear carbon chain such as:

zeta-Carotene
Phytoene (colorless)
Lycopene (red), E160d
Neurosporene
Phytofluene

  • cyclic carotenes: containing one or two cyclic structures such as:

alfa-Carotene (orange)
beta-Carotene (orange), E160a
gamma-Carotene (orange)
delta-Carotene
alfa-Zeacarotene
beta-Zeacarotene

  • hydroxycarotenoids (or carotenols): containing at least an hydroxyl group (xanthophylls) such as:

alfa-Cryptoxanthin (yellow)
beta-Cryptoxanthin (orange)
Lutein (yellow), E161b
Lycofill
Lycoxanthin
Rubixanthin
Zeaxanthin (yellow-orange)
Zeinoxanthin

  • epoxycarotenoids: containing at least an epoxic group (xanthophylls) such as:

Antheraxanthin
Auroxanthin
beta-Carotene-5,6-epoxide
Lutein-5,6-epoxide
Luteoxanthin
Neoxanthin
Violaxanthin (yellow)

  • uncommon or species-specific carotenoids such as:

Bixin, E160b
Capsanthin, E160c
Capsorubin, E160c
Crocetin

Note: although green leaves contain unesterified hydroxycarotenoids, most carotenols in ripe fruits are esterified with fatty acids. However, those of some fruits, particularly those that remain green when ripe (example kiwi fruit) undergo no or limited esterification.

Apocarotenoids

Apocarotenoids are a class of carotenoids containing less than 40 carbon atoms, very widespread in nature and with extremely different structures.
They derive from 40 carbon atom carotenoids by oxidative cleavage that can occurs through non-specific mechanisms, such as photo-oxidation, or through the action of specific enzymes (these enzymatic activities, identified in plants, animals and microorganisms, are collectively referred to as carotenoid cleavage dioxygenases).
Some of the most well-known

  • vitamin A
  • abscisic acid
  • bixin, E160b
  • crocetin
  • trans-β-apo-8′-carotenal, E160e
References

Boileau A.C., Merchen N.R., Wasson K., Atkinson C.A. and Erdman Jr J.W. cis-Lycopene is more bioavailable than trans-lycopene in vitro and in vivo in lymph-cannulated ferrets. J Nutr 1999;129:1176-1181 [Abstract]

de la Rosa L.A., Alvarez-Parrilla E., Gonzàlez-Aguilar G.A. Fruit and vegetable phytochemicals: chemistry, nutritional value, and stability. 1th Edition. Wiley J. & Sons, Inc., Publication, 2010

Engelmann N.J., Clinton S.K., and Erdman Jr J.W. Nutritional aspects of phytoene and phytofluene,carotenoid precursors to lycopene. Adv Nutr 2011:2;51-61 [Abstract]

Olempska-Beer Z. Lycopene (synthetic): chemical and technical assessment (CTA). Office of Food Additive Safety, Center for Food Safety and Applied Nutrition. U.S. Food and Drug Administration. College Park, Maryland, USA [PDF]

Periago M.J., Bravo S., García-Alonso F.J., and Rincón F. Detection of key factors affecting lycopene in vitro accessibility. J Agr Food Chem 2013;61(16):3859-3867 [Abstract]

Ross A.B., Thuy Vuong L., Ruckle J., Synal H.A., Schulze-König T., Wertz K., Rümbeli R., Liberman R.G., Skipper P.L., Tannenbaum S.R., Bourgeois A., Guy P.A., Enslen M., Nielsen I.L.F., Kochhar S., Richelle M., Fay L.B., and Williamson G. Lycopene bioavailability and metabolism in humans: an accelerator mass spectrometry study. Am J Clin Nutr 2011;93:1263-73 [Abstract]

Wang X-D. Lycopene metabolism and its biological significance. Am J Clin Nutr 2012:96;1214S-1222S [Abstract]