Co-NZyme R

Scientific Literature Review

August 2003 / rj

General Comment

This Scientific Literature Review has been done in August 2003 and covers
Biotin in relation to skin care. The scientific literature data bases used was
SciFinder among others. In this review the abstracts of the articles as well as
the source are listed. A selection of full articles are added at the end.

The information given in this review is based on our current knowledge and
experience, and may be used at your discretion and risk. We can not guarantee
this list to be complete and therefore do not assume any liability in connection
with this information.

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Transport of biotin in human keratinocytes
Grafe, Franziska; Wohlrab, Wolfgang; Neubert, Reinhard H.; Brandsch, Matthias.
Biozentrum of the Martin-Luther-University, Membrane Transport Group, Institute of
Pharmaceutics and Biopharmaceutics, Martin-Luther-University Halle-Wittenberg, Halle,
Germany
Journal of Investigative Dermatology (2003), 120(3), 428-433

Abstract
Biotin is an essential micronutrient for normal cellular function, growth, and development.
Biotin deficiency leads to pathol., dermatol., and neurocutaneous manifestations in skin and its
appendages. Previous studies described the presence of specific biotin transport systems in
the epithelia of the intestine, liver, kidney, and placenta, and in blood mononuclear cells. The
aim of this study was to examine biotin transport into human keratinocytes. Uptake of
[3H]biotin was measured both in the HaCaT cell line and in native keratinocytes in primary
culture. Uptake of [3H]biotin (6 nM) in HaCaT cells was linear for up to 5 min of incubation. In
the presence of an Na+ gradient total biotin uptake was 4- to 5-fold higher than in the absence
of sodium ions. Biotin uptake was not altered by H+ and Cl- gradients. This transport system
exhibited a Michaelis-Menten const. for biotin of 22.7 1.0 M and a maximal velocity of
163.6 3.5 pmol per 5 min per mg protein. [3H]Biotin uptake (6 nM) was strongly inhibited by
lipoic acid (oxidized form, Ki = 4.6 M; reduced form, Ki = 11.4 M), pantothenic acid (Ki = 1.2
M), and desthiobiotin (Ki = 15.2 M), but not by biocytin or biotin Me ester. Measured at
[3H]biotin concns. of 0.1-10 nM we obtained kinetic evidence for the presence of a second
transport component that is saturable at very low biotin concns. (Kt = 2.6 0.1 nM). Unlabeled
lipoic acid and pantothenic acid (20 nM) did not inhibit the [3H]biotin uptake (1 nM). We
conclude that human keratinocytes express the Na+-dependent multivitamin transporter with
preference for pantothenate and a very high affinity transport component with specificity for
biotin.

Applications of biotin
Lu, Ling; Chang, Champion; Shiu, Chih-Sheng; Lai, Jun-Liang; Kuo, Chih-Wei; Liu, Wen-Hu;
Shen, Yu-Ting; Hsu, Li-Lin; Tseng, Kuan-Chieh; Ho, Ko-Chieh. Labeltek Inc., Taipei,
Taiwan
Huaxue (2002), 60(4), 573-598 (General Review written in Chinese)

Abstract
A review. Biotin is a water-sol. vitamin generally classified as a B-complex vitamin. It helps in
the synthesis of fatty acids, in energy metab., and in the synthesis of amino acids and glucose.
Biotin is nowadays an important compd. that is-used as additives to food and cosmetics,
vitamin, health foods and as labeling agent with proteins and nucleic acids. Biosynthesis
pathway, chem. synthesis, daily allowance, deficiency, etc., of biotin were also discussed. The
aim of this article is to review the recent successful applications of biotin in a no. of industrial
products.

Microbial processes for the production of organic sulfur compounds as

cosmetic materials
Kino, Kuniki. Department of Applied Chemistry, School of Science and Engineering, Waseda
University, Tokyo, Japan
Fragrance Journal (2003), 31(3), 16-22 (General Review written in Japanese)

Abstract
A review on microbial processes for the prodn. of the sulfur compds. such as L-cysteine,
glutathione, and biotin which are useful cosmetic materials.

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Biotin in microbes, the genes involved in its biosynthesis, its biochemical role
and perspectives for biotechnological production
Streit, W. R.; Entcheva, P. Institut fuer Mikrobiologie und Genetik, Universitaet Goettingen,
Germany
Applied Microbiology and Biotechnology (2003), 61(1), 21-31

Abstract
A review. Biotin (vitamin H) is one of the most fascinating cofactors involved in central
pathways in pro- and eukaryotic cell metab. Since its original discovery in 1901, research has
led to the discovery of the complete biotin biosynthesis pathways in many different microbes
and much work has been done on the highly intriguing and complex biochem. of biotin
biosynthesis. While humans and animals require several hundred micrograms of biotin per
day, most microbes, plants and fungi appear to be able to synthesize the cofactor themselves.
Biotin is added to many food, feed and cosmetic products, creating a world market of 10-30
t/yr. However, the majority of the biotin sold is synthesized in a chem. process. Since the
chem. synthesis is linked with a high environmental burden, much effort has been put into the
development of biotin-overproducing microbes. A summary of biotin biosynthesis and its biol.
role is presented; and current strategies for the improvement of microbial biotin prodn. using
modern biotechnol. techniques are discussed.

Biotin-dependent carboxylase activities in different CNS and skin-derived cells,

and their sensitivity to biotin-depletion
Suormala, Terttu; Wiesmann, Ulrich N.; Cruz, Fatima; Wolf, Armin; Daschner, Markus; Limat,
Alain; Fowler, Brian; Baumgartner, E. Regula. Metabolic Unit, University Children's Hospital,
Basel, Switz.
International Journal for Vitamin and Nutrition Research (2002), 72(4), 278-286

Abstract
The validity of various transformed and untransformed CNS and skin-derived cell cultures as a
model for studying effects of biotin deficiency was tested. In biotin-sufficient conditions (0.1-10
mol/L) all cell types showed considerable activities of the four biotin-dependent
carboxylases. Notably, pyruvate carboxylase activity was also present in the different
neuronal cells. One passage in low-biotin medium (6-130 pmol/L) lowered mitochondrial
carboxylase activities in all cell types, but to varying degrees. Sensitivity to biotin depletion
was greatest in three neuronal cell types, Roc-1 oligodendroglia, and three keratinocyte cell
types (carboxylase activities decreased to 2-11% of maximal); intermediate in primary
astrocytes and C6 glioma (decreased to 12-28%), and least in SAOS2 sarcoma and skin
fibroblasts (decreased to 32-85%). Transformed and untransformed cell lines of the same cell
type showed similar sensitivity. We conclude that cultures of different transformed CNS and
keratinocyte cell types allow the study of effects of biotin deprivation. Carboxylase activities of
neurons, oligodendroglia, and keratinocytes were much more sensitive to biotin depletion than
fibroblasts. This may be an important factor in the pathogenesis of neurol. and cutaneous
abnormalities in congenital biotinidase deficiency where recycling of biotin is deficient.

Final report on the safety assessment of biotin

Fiume M Z, Cosmetic Ingredient Review Expert Panel, USA.
International Journal of Toxicology (2001), 20(Suppl. 4), 1-12

Abstract
A review. Biotin is a water-sol. vitamin used as a hair-conditioning agent and a skin-
conditioning agent in many cosmetic products at concns. ranging from 0.0001% to 0.6%.
Although biotin does absorb some UV radiation, the absorption shows no peaks in the UVA or
UVB region. Biotin is rapidly metabolized and excreted in urine. Little acute oral toxicity is

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seen in animal tests. Short-term and subchronic toxicity studies likewise found no evidence of
toxicity. Although intradermal injection of a small quantity of biotin (0.1 mL) into guinea pig
skin did not produce skin irritation, biotin (0.1% at pH 7.3) did produce slight, transient ocular
irritation in rabbit eyes. Biotin was not mutagenic in bacterial tests, but pos. results were found
in a Tradescantia micronucleus test. There was evidence of an increase in the no. of
resorptions in rats receiving biotin by s.c. injection, with concomitant decreases in fetal,
uterine, and placental wts. Another study of mice receiving biotin orally or by s.c. injection
found no differences between control and treatment groups. Although there is one case study
reporting an urticarial reaction in the literature, there are a very large no. of individuals
exposed to biotin on a daily basis, and there is not a parallel appearance of irritation,
sensitization, or other adverse reactions. Based on these available data, it was concluded that
biotin is safe as used in cosmetic formulations.

Percutaneous absorption of biotin in healthy subjects and in atopic dermatitis

patients
Makino Y; Osada K; Sone H; Sugiyama K; Komai M; Ito M; Tsunoda K; Furukawa Y Makino
Dermatology Clinic, Sendai, Japan
Journal of nutritional science and vitaminology (1999 Jun), 45(3), 347-52

Abstract
The study was designed to test the ability of sequential applications of biotin-containing
ointment to increase serum biotin levels. Twenty atopic dermatitis patients (mean age, 20.5
yr) and 11 healthy subjects (mean age, 25.5 yr) volunteered to participate in this study. The
diagnosis of atopic dermatitis was established dermatologically. Seven grams per day of
ointment containing 0.3% biotin and 1-4 g per day of steroid ointment were both applied
sequentially. The healthy subjects applied only biotin ointment. The biotin concentration was
determined microbiologically. Before biotin treatment, the average serum biotin level was
significantly lower in atopic dermatitis patients than in healthy subjects. The percutaneous
application of biotin-containing ointment caused a significant increase in the serum biotin
concentration in both healthy subjects (from 41.5 +/- 10.0 to 50.2 +/- 9.2 nmol/L) and in atopic
dermatitis patients (from 27.9 +/- 17.4 to 50.7 +/- 21.6 nmol/L), especially in patients whose
initial level was low, and also could be effective in regulating the atopic allergic response
involving eosinophils. In conclusion, biotin appears to be readily absorbed through both
normal and dermatitis-affected human skin.

Rapid quantification of biotin in human skin extracts after dermal application

using high-performance liquid chromatography-electrospray mass spectrometry
Wolf, Raik; Huschk, Christoph; Raith, Klaus; Wohlrab, Wolfgang; Neubert, Reinhard. Dep.
Pharmacy, Inst. Pharmaceutics Biopharmaceutics, Martin Luther Univ. Halle-Wittenberg,
Halle (Salle), Germany
Analyst (Cambridge, United Kingdom) (1997), 122(11), 335-337

Abstract
A method for the detn. of biotin in exts. from different layers of human skin is presented, which
has a detection limit of 800 pg mL-1 without further sample prepn. The quantification was
performed by using an HPLC procedure on a reversed-phase column, which takes only 4 min,
coupled to a single quadrupole mass spectrometer via an electrospray ionization interface.
The detection was performed in the pos. selected ion monitoring mode. Addnl. MS-MS expts.
with biotin using an ion trap mass spectrometer are described.

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Rapid Quantification of Biotin in Human Skin Extracts After Dermal Application
Using High-performance Liquid Chromatography-Electrospray Mass
Spectrometry
Wolf, Raik; Raith, Klaus; Neubert, Reinhard; Huschka, Christoph; Wohlrab, Wolfgang.
Institute of Pharmaceutics and Biopharmaceutics, Department of Pharmacy, Martin Luther
University Halle-Wittenberg, Halle (Saale), Germany
Analytical Communications (1997), 34(11), 335-337

Abstract
A method for the detn. of the vitamin biotin in exts. from different layers of human skin is
presented, which has a detection limit of 800 pg ml-1 without further sample prepn. The
quantification was performed using an HPLC procedure on a reversed phase column, which
takes only 4 min, coupled to a single quadrupole mass spectrometer via an electrospray
ionization interface. The detection was performed in the pos. selected ion monitoring mode.
Addnl. MS-MS and MSn expts. with biotin using an ion trap mass spectrometer are described.

Vehicle-dependent biotin liberation and penetration into human skin

Wagner, C.; Gobel, S.; Wohlrab, W.; Neubert, R. Klinik Poliklinik Hautkrankheiten, Martin-
Luther-Univ., Halle/Saale, Germany
Skin Pharmacology (1997), 10(1), 34-39

Abstract
The influence of the vehicle was investigated on biotin release and penetration after topical
application. Liberation studies (multi-layer membrane model) showed that >50% of the biotin
in an oil-in-water (O/W) emulsion and a microemulsion (ME) was released within 300 min,
whereas the degree of release from a water-in-oil emulsion did not exceed 16%. For this latter
vehicle, the influence of a controlled drug release on the penetration processes in human skin
(Franz cell) resulted in the penetration of only small quantities of the drug into the skin layers
(horny layer 7.9, living epidermis 0.11, dermis 0.38% within 300 min.). The application of an
O/W emulsion or an ME lead to a large reservoir in the horny layer (O/W 26.5, ME 26.0%
within 300 min.) and to important epidermal and dermal concns. (living epidermis: O/W 2.0,
ME 0.3%; dermis: O/W 3.4, ME 1.6% within 300 min.). The time-dependent concn. profiles in
the skin layers suggest that the use of an ME produces an immediate effect, whereas the
application of an O/W emulsion results in a delayed and/or long-term effect.

Proliferation and differentiation of cultured human follicular keratinocytes are

not influenced by biotin
Limat, Alain; Suormala, Terttu; Hunziker, Thomas; Waelti, Ernst R.; Braathen, Lasse R.;
Baumgartner, Regula. Dermatological Clinic, Inselspital, Bern, Switz
Archives of Dermatological Research (1996), 288(1), 31-8

Abstract
In humans and in animals, biotin deficiency causes pathol. changes in the skin and its
appendages. High doses of biotin may also have beneficial effects on skin, hair and
fingernails in humans and animals with normal biotin status. Therefore, we investigated the
effects of low and high concns. of biotin on proliferation and differentiation of cultured outer
root sheath cells from human hair follicles as an in vitro model for skin. The activities of biotin-
dependent carboxylases were measured to evaluate the biotin status of the cells. In
monolayer cultures of outer root sheath cells, proliferation and expression of the
differentiation-specific keratins K1 and K10 were not influenced by extremely low concns. of
biotin (< 2 10-10 mol/L) or by pharmacol. doses of biotin (10-5 mol/L). Biotin deficiency of
the cells was confirmed under the former condition by demonstrating decreased activities of
the mitochondrial carboxylases. In organotypic cocultures of outer root sheath cells and

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dermal fibroblasts, in which stratified epithelia resembling epidermis were developed, the biotin
concn. had no effect on the expression of all tested epidermal differentiation markers,
including the suprabasal keratins K1 and K10, the hyperproliferation-assocd. keratin K16,
involucrin and filaggrin.

Influence of Topically Applied Biotin on fine Wrinkles in the Skin of Elderly

Patients
L.Gilli, G.L.Floersheim
Zeitschrift f. Hautkrankheiten 70(6), (1995) 419-425 (written in German)

Abstract
Demonstration of a distinct reduction of fine wrinkles in the skin of elderly persons.

Recent topics of plant extracts and their applications to cosmetics

Kushibashi, Kengo; Yamaki, Hidehiko. Koei Perfum. Co., Ltd., Tokyo, Japan
Fragrance Journal (1994), 22(2), 54-61

Abstract
A review with 14 refs. on metab. and physiol. activities of biotin, biotin contents in herbs, and
application of their exts. to cosmetics.
Pharmacologic effects of biotin on epidermal cells
Fritsche A.; Mathis G. A.; Althaus F. R.
Schweizer Archiv fr Tierheilkunde (1991) 133(6), 277-83 (written in German)

Abstract
Biotin deficiency in animals causes pathological changes of the skin and its appendages
including, for example, exfoliative dermatitis, depigmentation, and alopecia. The hooves of
biotin-deficient swine are weak, brittle, and often necrotic. These changes disappear after
dietary biotin supplementation. Biotin supplementation also noticeably improves the hoof
quality of horses, cattle and swine having no apparent biotin deficiency. In order to elucidate
the molecular basis of these effects, the influence of biotin on cytokeratin expression in a
keratinocyte cell line (Ha-CaT) was investigated using electrophoretic and immunological
techniques. Pharmacological biotin concentrations of 1 microM, and 100 microM in the culture
medium caused a specific increase in cytokeratins, which are normally induced upon terminal
differentiation of epidermal cells in vivo. The expression of cytokeratins occurring in stratified
epithelia independent of differentiation were not affected. These findings show that biotin
directly stimulates the differentiation of epidermal cells. Such a molecular mechanism
revealed in cell culture could provide an explanation for the therapeutic effects of
pharmacological doses of biotin on hoof quality in farm animals.

Skin manifestations of biotin deficiency

Mock D M Department of Pediatrics, University of Iowa Hospitals and Clinics, Iowa City
52242
Seminars in Dermatology (1991 Dec), 10(4), 296-302

Abstract
This article reviews current knowledge concerning the dermatologic manifestations of biotin
deficiency. Biotin is a water-soluble vitamin that acts as an essential cofactor for four
carboxylases, each of which catalyzes an essential step in intermediary metabolism. For
example, acetyl-CoA carboxylase catalyzes the rate-limiting step in fatty acid elongation. In
infants, children, and adults, deficiency of biotin causes alopecia and a characteristic scaly,
erythematous dermatitis distributed around body orifices. The rash closely resembles that of

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zinc deficiency. Candida albicans often can be cultured from the skin lesions. Biotinidase
deficiency, an inborn error, causes biotin deficiency, probably as a consequence of unpaired
intestinal absorption, cellular salvage, and renal reclamation of biotin; biotinidase deficiency
causes dermatologic manifestations similar to biotin deficiency. There is evidence that
impaired fatty acid metabolism secondary to reduced activities of the biotin-dependent
carboxylases (especially acetyl-CoA carboxylase) plays an etiologic role in the dermatologic
manifestations of biotin deficiency. Candida infections secondary to impaired immune function
might also contribute to the dermatitis of biotin deficiency.

Vitamins in cosmetics
Erlemann, Gustav. Dep. VL, TS Hoffman-La Roche, Fed. Rep. Ger.
Aerosol & Cosmeticos (1985), 7(38), 4, 6, 8-11 (Journal written in Portuguese)

Abstract
Ascorbyl palmitate [137-66-6] had an antioxidant activity in cosmetics, effect enhanced by
lecithin and/or tocopherol. The highest activity was shown by Ronoxan A (tocopherol-ascorbyl
palmitate-lecithin mixt.) [83868-85-3]. In other expts., the soly. of biotin [58-85-5] and of its
derivs. in water and aq. EtOH was detd. Biotin Et ester [87573-52-2] showed a higher soly. in
aq. EtOH than biotin and a better skin absorption in humans. Vitamins used as additives to
cosmetics are discussed.

Biotin; proposed affirmation of GRAS status

United States Food and Drug Administration, Rockville, MD, USA.
Federal Register (1983), 48(10), 1739-42

Abstract
biotin [58-85-5] Is proposed to be generally-recognized-as-safe (GRAS) as a direct ingredient
of food, under the Federal Food, Drug, and Cosmetic Act. Biotin is used as a nutrient
supplement, and in special formulas for wt. redn. and gain and in infant formulas.

Cosmetic application of vitamins

Hennies, Klaus
Riechstoffe, Aromen, Koerperpflegemittel (1973), 23(8), 241-2, 244

Abstract
The beneficial effect of vitamin-contg. cosmetics depends on the ability of the body skin to
absorb the vitamin, so that they can pass into the blood stream to carry out their functions.
Skin absorbs vitamins such as A, D2, E, H, B6, and C, applied in the form of water or oil sol.
prepns.

Additives which enhance the efficiency of cosmetics

Sherr-Bruzer, Emilia. Hung.
Olaj, Szappan, Kozmetika (1972), 21(1), 18-21, (General Review written in Hungarian)

Abstract
A review of biol. active substances as additives in cosmetic prepns. The roles and effects of
A-, B1-, B2-, B12-, H-, D-, E-, and F-vitamins, pantothenic acid, vegetable (corn and wheat
germ) oils, hamamelis ext., chamomile ext., birch ext., peloids, pollen, fruit juices, camphor,
menthol, chlorophyll, azulene, allantoin, capsaicin, antiseptics (boric acid, tribromo-
salicylanilide, hexachlorophene) and some antioxidants used frequently in the cosmetic
industry are described. 7 refs.

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Vitamins in cosmetics
DeRitter, E.; Magid, L.; Sleezer, P. E.
American Perfumer and Aromatics (1959), 73(No. 5), 54-8

Abstract
Topically applied vitamins (A, D2, E, biotin, panthenol, and pyridoxine) and the incorporation in
cosmetics of the fat-sol. and water-sol. vitamins are discussed. The topical application of
vitamins can provide a local concn. in the skin higher than is obtained by oral administration
and a beneficial pharmacodynamic effect. Formulation techniques are outlined. 73
references.