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`Ann. Nutrit. Alimentation Vol. 19 pp. pages C655-C658 (1965)
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`THE STUDY OF ASTAXANTHIN TRANSFORMATION INTO VITAMIN A IN THE
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`ALBINO RAT:
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`IN VITRO EXPERIMENTS
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`by
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`Ms. R. MASSONET, Mrs. T. CONQUY and R. R. GRANGAUD
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`Laboratory of Medical Biochemistry in the Mixed Faculty of Medicine and Pharmacy, Rennes.
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`It is traditionally thought that the only substances in mammals which display vitamin A properties are
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`those with an axerophthol group formed by the union of a [3-ionone nucleus and an isoprenic chain,
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`i.e. a potent molecule of vitamin A. Carotenoid pigments, with 40 atoms of carbon in which two
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`nuclei are oxygenated, such as zeaxanthin, xanthophyll etc. are deprived of vitamin A activity.
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`However, astaxanthin, 3.3’-dihydroxy 4.4’-diceto-[3-carotene, the main carotenoid pigment in
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`crustaceans, constitutes a singular exception in this regard. Administered to the vitamin A deficient
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`white rat, this pigment manifests a notable antixerophthalmic activity, however, it is distinct from
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`other vitamin A factors since significant weight gain is only observed with doses much higher than
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`those which can heal ocular lesions (2) (3) (1). In addition, regardless of the administered pigment
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`dose, serious disorders in the reproduction functions are noticed: males become sterile and, while
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`females can still be impregnated, gestation is profoundly disturbed (miscarriages, still-births [9]).
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`C655
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`
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`Nevertheless, research conducted at the same time in fish, should show that the latter are able to use
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`astaxanthin as a vitamin A precursor. In fact, the administration of the pigment to Gambusia
`holbrooki Grd. leads to a neoformation of retinol which is detectable in the intestinal mucosa, the
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`liver and the eyes (5) (4) (7) (12).
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`Examination of this data set suggests that the rat may have the same ability but only in a localized
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`manner and in a quantitatively-limited manner. This hypothesis, subjected to experimentation,
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`received an initial confnmation (6) (10): after administration of astaxanthin diacetate, the rat was
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`incapable of creating pigment reserves but the content of retinol in the retina was notably increased. In
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`vitro experimentation shows that the ocular tissue performs the astaxanthin to vitamin A conversion.
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`This research was pursued in order to specify both the chemical and tissue specificity. Comparatively,
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`astaxanthin acetate, [3-carotene and xanthophyll dipalmitate were incubated with retinal, intestinal
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`mucous and adrenal tissue, successively, from deficient rats. The extraction of astaxanthin, the
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`preparation of diacetate and the purity controls were previously described as well as the preparation of
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`animals and the incubation techniques (10).
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`Animals were sacrificed by decapitation, three days after the appearance of the deficiency symptoms
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`(weight stabilization and beginning of xerophtalmy). The organs were immediately removed.
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`The eyes (except for the first experiment which was performed on the intact ocular globe) were
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`vertically sectioned in order to only preserve the retinal region which was immediately immersed in
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`the incubation liquid. In order to eliminate any error due to individual variations, two lots were
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`created by combing the right eye segments of half of the subjects (seven) with the left eye segments of
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`the remaining animals and vice versa, the incubation liquid of the first lot being the only lot diluted
`with carotenoid.
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`For the creation of the adrenal lots, the same precautions were observed.
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`The intestines were washed using 9% physiological serum and then cut into 1 cm sections. Each of
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`the two lots was composed of six alternate sections.
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`In all the experiments, the incubation time at 37C was uniformly kept to 3 ‘/2 hours except for the first
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`(whole eyes) which were kept for 12 hours.
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`At the end of this time, the tissue fragments from the various lots were saponified in accordance with
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`the Lewis, Bodansky, Falk and MacGuire technique (8). The unsaponifiable [tissues] were treated
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`with petrol ether. After washing and evaporation in a nitrogen atmosphere under reduced pressure, the
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`residue was dissolved in chloroform. The search and dose of vitamin A were performed using the Carr
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`and Price reaction in accordance with the Meunier and Raoul technique (11).
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`The results are recorded in the following table:
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`C656
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`Vitamin A in ;
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`1.65
`0.75
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`C’?‘mo
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`Incubated Oran
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`Whole eyes
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`Retinal region of eyes
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`Retinal region of eyes
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`Retinal region of eyes
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`Adrenal [tissue]
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`Adrenal [tissue]
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`Adrenal [tissue]
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`Intestines
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`Carotenoid Addition
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`Astaxanthin diacetate
`0
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`Astaxanthin diacetate
`0
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`-carotene
`0
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`Xantho h ll dialmitate
`0
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`Astaxanthin diacetate
`0
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`5 -carotene
`0
`Xantho h ll di almitate
`0
`Astaxanthin diacetate
`-carotene
`0
`* Non-dosable
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`It clearly appears that, in the rat, it’s the retinal tissue which alone has the ability to convert
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`astaxanthin into retinol. The absence of any xanthophyll transformation also highlights the fact that, in
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`accordance with the traditional data, this property does not extend to all carotenoids whose two cycles
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`are oxygenated.
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`The limitation of this ability to convert astaxanthin in the retinal tissue indicates the essentially
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`antixerophthalmic activity which this carotenoid manifests in the rat. Vitamin A forms in local
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`quantities which are not sufficient to allow its passage into circulation, the action on the general status
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`is, at least partially abolished, from where the dissociation observed in the pigment’s vitamin activity.
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`C657
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`BIBLIOGRAPHY
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`(1) R.Grangaud, Doctoral Thesis in Physical Sciences, Lyon, 1950.
`(2) R.Grangaud and R. Massonet, C.R.Acad.Sc., 1948, V.227, p.568.
`(3) R.Grangaud and R. Massonet, C. R. Acad. Sc., 1950, V.230, p.1319.
`(4) R.Grangaud and R. Mnssonet, Arch.Sci.physiol., 1955, V.9, p.3.
`(5) R.Grangaud and R. Massonet, C.R.Acad.Sc., V241, 1955, p. 1087.
`(6) R.Grangaud, R.Massonet, Th.Conquy, and J.Ridolfo, C. R. Acad. Sc., 1961, V252, p.1854.
`(7) R.Grangaud, P.Vignais, R. Massonet, and J.P.Moatti, Bull.Soc. chim. Biol., 1957, V.39, no.11.
`(8) J.M.Lewis, O.Bodansky, K.G.Falk and G. MacGuire, .1. Nutrition, v. 23, 1941, p.351.
`(9) R. Massonet, Doctoral Thesis in Sciences, Lyon, 1958.
`(10) R.Massonet, T.Conquy, and R.Grangaud, C. R. Acad. Sc., 196[1], V.155, p.747.
`(11) P.Meunier and Y.Raoul, Diagnostic chemistry of Vitamin Deficiencies. Masson and Co., Paris
`1942.
`
`(12) J.P.Moatti, Doctoral Thesis (Pharmacy), Algiers, 1959.
`
`DISCUSSION
`
`Mr. Raoul. — I observed that the retina is added to already-known areas where a double-type
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`carotenoid is transformed into a single-type: intestinal mucosa and liVer where the marked [3-carotene
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`was used for this trial. Recent work with marked [3-carotene has confirmed the effectiveness of the
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`hepatic parenchyma. There are, therefore, many possibilities.
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`Mr. Matet. — 1. Did the author find alcohol Vitamin A or retinal Vitamin A in the eyes of the rats
`which receiVed astaxanthin?
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`Ms. Massonet. — In the examined animals, it was retinol which was found.
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`Mr. Matet. — 2. In the same animals, was unaltered astaxanthin found, especially in the liVer?
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`Ms. Massonet. — In the animals treated with astaxanthin, the pigment was found, in trace amounts, in
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`the eyes and thyroids. The pigment was missing in the other organs examined and, in particular, it was
`neVer detected in the liVer.
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`Mr. Grangaud. — The 1948 publication of the obserVation of the exclusiVely antixerophthalmic
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`properties of penaeid oils was generally and naturally receiVed with skepticism. HoweVer, Paul
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`Meunier agreed with our results and encouraged us to continue. Also, the completion of this work
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`which has just been presented to you is, in my opinion, an homage to his memory and I am happy to
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`declare it here in front of many of his closest friends and collaborators.
`
`C658
`
`
`
`The Language Link of Connecticut
`Translation, DTP and Foreign Language Typesetting
`
`I hereby certify to the best of my knowledge and ability that the six translations referenced below are true
`and accurate translations from French to English of the French language originals and were performed by
`a professional translator with over 10 years translation experience.
`
`The above facts are personally known to me or are based on representations made tome by the
`translator.
`
`
`
`
`
`
`TITLE: Antixerophthalmic effect of the esters of astxanthln
`Authors: GRANGAUD, R.; MASSONET, R.
`Journal: Comptes rendus des seanoes de la Soclete de biologie 1954 Vol. 148
`pp. 1392-1394
`
`Date final
`translated
`version a a roved
`May 23, 2013
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`
`
`Author, Title and Journal
`
`Our file name
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`
`
`rangaud 1954 CRdesSc blologle
`Vol. 143 pp. 1392-1394 EN
`18May2013.docx
`
`
`
`Title: Transformation of astaxanthin to vitamin A by ocular tissue of the rat in
`
`vitro.
`Authors: MASSONET, R.; CONQUY, T.; GRANGAUD, R.
`Author Affiliation: Lab. Chim. blol., Fac. Med. Pharrn., Algiers.
`Joumalz Compte rendu des seanoes de la Societe de biologie 1961 Vol. 155
`pp. 747-750
`
`Grangaud 1961a_ CRdesSc
`blologlevol. 155 pp. 747-750 EN
`18May2013
`
`
`
`
`Title: Transformation of astaxanthin to vitamin A in the albino rat:
`Grangaud 1961b_ CRdesSc Vol. 252
`
`
`neoformation in vivo and in vitro.
`'
`pp. 1854-1856 EN 18May2013
`Authors: GRANGAUD, Rene; MASSONET, Renée; CONQUY, Thérése:
`
`RIDOLFO, J.
`Author Affiliation: Lab. Chim. biol., Fac. Med. Pharm.. Algiers.
`Journal: Comptes Rendus de l'Academie des Sciences 1961 Vol. 252 pp.
`18541856
`
`
`
`May 23, 2013
`
`Title: The study of astaxanthin transformation into vitamin A in the albino rat:
`In vitro experiments
`Authors: MASSONET. R.; CONQUY, T.; GRANGAUD, R.
`Author Affiliation: Lab. Chim. bioI., Fac. Med. Pharm., Rennes
`Journal: Compte rendu des seances de la Societe de biologie 1965
`Pages-C655~C658
`
`Grangaud 1965 Ann. Nutrit.
`Alimentation Vol. 19 pp. 0655-0658
`EN 18 May2013
`
`May 23, 2013
`
`TITLE: The Action of Shrimp Oil (Penaeus foliaoeus) on the Vitamin A
`Deficient White Rat
`Authors: GRANGAUD, Rene.; MASSONET. Renee.
`Journal: Comptes rendus de l'Academie des Sciences 1948 Vol. 227
` . 568-570
`May 23, 2013
`TITLE: Antixerophthalmic Activity of the Carotenoid Pigment of the
`Grangaud 1950 CRdesSc V230
`Arlsteomorphafollacea (Peneida)
`pp1319-1321 EN 1BMay2013
`Authors: ‘GRANGAUD, Rene.; MASSONET. Renee.
`Journal: Comples Rendus de l‘Academie des Sciences, March 27, 1950 Vol.
`230 pp. 1319-1321
`
`
`Grangaud 1948 CRdesSc V227
`pr>568-570 EN 181/1672013
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