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Figure 1. Biotin, C₁₀H₁₆N₂O₃S, Mw = 224,31 g/mol Biotin occurs in foods as the free vitamin or bound to protein. Syn..."

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Figure 1. Biotin, C10H16N2O3S, Mw = 224,31 g/mol

Biotin occurs in foods as the free vitamin or bound to protein.

Synonyms for biotin:

* vitamin B7, co-enzyme R, vitamin H

[https://metrofood-wiki.foodcase-services.com/index.php?title=Summary+-+biotin Summary]

-----

=== Golden standard ===

[http://www.cen.eu/cen/Sectors/TechnicalCommitteesWorkshops/CENTechnicalCommittees/Pages/Standards.aspx?param=6256&title=CEN/TC%20275 EN 15607:2009] Foodstuffs - Determination of d-biotin by HPLC

=== Method indicator ===

* ''Name''
* ''Code''

=== Scope ===

The European Standard EN 15607 specifies a method for the determination of the mass fraction of d-biotin in foodstuffs by high performance liquid chromatography (HPLC). This method can also be used for estimation of d-biocytin, but it has to be noted that the method has not been validated for the determination of d-biocytin.

=== Principle ===

Pretreatments include enzymatic treatment followed by extraction. Quantification is done by HPLC with post-column binding reaction. Post-column reaction of d-biotin with avidin protein that is bound to a fluorescent marker, provides a specific method.

=== Key steps ===

==== Extraction ====

* The sample should be homogenised prior to extraction.
* Gluthatione solution, EDTA solution, citrate buffer solution and papain solution are added.
** Use of taka-diastase may be needed if the sample contains high amounts of starch.
* The sample solution is incubated overnight (37 °C).
* The solution is filtered. 

==== HPLC ====

* Liquid chromatographic system consisting of a pump, an injector, a fluorescence detector (excitation wavelength 490 nm and emission wavelength 520 nm) and a data evaluation system.
* Reverse phase column: particle size 5 µm, diameter 4,0 mm, length 250 mm
* Mobile phase: phosphate buffer (pH 6):methanol (8:2, v:v)
* Also other chromatographic systems can be used if equivalent results are guaranteed.
* The performance criteria is the baseline separation of d-biotin from interferences.
* The system should include a post-column reactor derivatisation unit with a suitable reagent delivery system.
** d-biotin is derivatised with fluorescent avidin-fluorescein 5-isothiocyanate.

 

==== Identification and detection ====

* D-biotin is detected fluorometrically (excitation wavelength 490 nm, emission wavelength 520 nm)
* Identification of d-biotin is done by the comparison of the retention time obtained with the standard test solution to that of the sample test solution.
** Identification can also be done by adding the standard solution to the sample test solution. 

 

==== Quantification and calculations ====

* Quantification is done by external calibration.
* Peak areas or heights are integrated and second degree calibration curve is used.
* The mass fraction of d-biotin in µg/100 g of the sample is calculated.

 

 

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=== Remarks ===

* Other methods for the determination of d-biotin:
** microbiological method using ''Lactobacillus plantarum''
** radio-assays using specific binding protein
* The method described in the standard EN 15607 can also be used for estimation of d-biocytin.

=== Criteria for analytical performance and analytical quality control ===

* Greenfield and Southgate discuss the criteria for analytical performance and quality of analytical data in their book.
** [ftp://ftp.fao.org/docrep/fao/008/y4705e/y4705e00.pdf Greenfield H, Southgate DAT. 2003. Food composition data ? production, management and use. Elsevier Applied Science, London, UK.]

* [https://metrofood-wiki.foodcase-services.com/index.php?title=Precision+data+-+biotin link to precision data]

=== Certified Reference Materials/Standard Reference Material ===

* [https://metrofood-wiki.foodcase-services.com/index.php?title=Reference+materials+-+biotin link to detailed data]

=== Proficiency testing schemes ===

Here are listed some completed, on-going and/or upcoming proficiency testing schemes concerning biotin:

[http://otis.jtlnet.com/~proficie/proficiencytesting.com.au/documents/651.pdf PTA - Proficiency Testing Australia: Report No. 651, Food Proficiency Testing Program, Round 31 - Vitamins]

[http://www.dla-lvu.de/english/DLA%20Programm%202012%20english.html DLA - Dienstleistung Lebensmittel Analytik GbR: 33/2012 Food Supplement II: Biotin, Vitamin C, Niacin and Pantothenic Acid]

Some upcoming proficiency testing schemes can be found in the [http://www.eptis.bam.de/en/index.htm EPTIS database].

=== Other methods available ===

 

=== Literature ===

* see separate child page below

== EuroFIR assistance to this method/guidelines ==

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 * Greenfield and Southgate discuss the criteria for analytical performance and quality of analytical data in their book.
-** [ftp://ftp.fao.org/docrep/fao/008/y4705e/y4705e00.pdf Greenfield H, Southgate DAT. 2003. Food composition data ? production, management and use. Elsevier Applied Science, London, UK.]
+** [ftp://ftp.fao.org/docrep/fao/008/y4705e/y4705e00.pdf Greenfield H, Southgate DAT. 2003. Food composition data - production, management and use. Elsevier Applied Science, London, UK.]
 *  [https://metrofood-wiki.foodcase-services.com/index.php?title=Precision+data+-+biotin link to precision data]

Biotin - Revision history

Admin: 4 revisions imported

Admin: Protected "Biotin" ([Edit=Allow only administrators] (indefinite) [Move=Allow only administrators] (indefinite))

Admin at 07:58, 23 November 2017

Admin at 07:26, 21 November 2017

Admin: Created page with " Figure 1. Biotin, C10H16N2O3S, Mw = 224,31 g/mol Biotin occurs in foods as the free vitamin or bound to protein. Syn..."

Admin: Created page with "

Figure 1. Biotin, C₁₀H₁₆N₂O₃S, Mw = 224,31 g/mol Biotin occurs in foods as the free vitamin or bound to protein. Syn..."