Testing for Biotin Interference in In 
Vitro Diagnostic Devices 
 
 
Guidance for Industry 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
For questions on the content of this guidance, contact Center for Biologics Evaluation and 
Research (CBER), Office of Communication, Outreach, and Development (OCOD) at 240-402-
8010 or 800-835-4709, or email ocod@fda.hhs.gov.  For questions about this document 
concerning products regulated by Center for Devices and Radiological Health (CDRH), contact 
the Office of In Vitro Diagnostics at 301-796-5900, or email CDRH-OIR-Policy@fda.hhs.gov. 
 
 
 
 
 
 
 
 
 
 
 
 
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Biologics Evaluation and Research 
Center for Devices and Radiological Health 
October 2020  Testing for Biotin Interference in In 
Vitro Diagnostic Devices 
 
 
Guidance for Industry 
 
 
Additional copies of this guidance are available from: 
 
Office of Communication, Outreach and Development 
Center for Biologics Evaluation and Research 
Food and Drug Administration 
10903 New Hampshire Ave., WO71, Room 3128  
Silver Spring, MD 20993 
Phone:  800-835-4709 or 240-402-8010 
Email:  ocod@fda.hhs.gov  
https://www.fda.gov/vaccines-blood-biologics/guidance-compliance-regulatory-information-
biologics/biologics-guidances 
or 
Office of Policy 
Guidance and Policy Development 
Center for Devices and Radiological Health 
Food and Drug Administration 
10903 New Hampshire Ave., WO66, Room 5431 
Silver Spring, MD 20993 
Phone:  301-796-5900 
https://www.fda.gov/medical-devices/device-advice-comprehensive-regulatory-
assistance/guidance-documents-medical-devices-and-radiation-emitting-products 
 
 
 
  Contains Nonbinding Recommendations 
Table of Contents 
 
 
I.  INTRODUCTION............................................................................................................. 1 
II.  BACKGROUND ............................................................................................................... 1 
III.  BIOTIN TESTING RECOMMENDATIONS ............................................................... 2 
IV.  REFERENCES .................................................................................................................. 4 
i 
  Contains Nonbinding Recommendations 
Testing for Biotin Interference in In Vitro Diagnostic Devices 
 
 
Guidance for Industry 
 
 
This guidance represents the current thinking of the Food and Drug Administration (FDA or 
Agency) on this topic.  It does not establish any rights for any person and is not binding on FDA 
or the public.  You can use an alternative approach if it satisfies the requirements of the 
applicable statutes and regulations.  To discuss an alternative approach, contact the FDA staff 
responsible for this guidance as listed on the title page. 
 
 
I.  INTRODUCTION 
 
The Food the Drug Administration (FDA or we) is providing recommendations on the testing for 
interference by biotin on the performance of in vitro diagnostic devices (IVDs).  This guidance is 
intended to help device developers and clinicians understand how FDA recommends biotin 
interference testing be performed, and how the results of the testing should be communicated to 
end-users, including clinical laboratories and clinicians.  The recommendations apply to IVDs, 
including devices that are licensed under section 351 of the Public Health Service Act (42 U.S.C. 
262) and used in donor screening, that use biotin technology.  This guidance finalizes the draft 
guidance of the same title dated June 2019.   
 
FDAs guidance documents, including this guidance, do not establish legally enforceable 
responsibilities.  Instead, guidances describe the FDAs current thinking on a topic and should be 
viewed only as recommendations, unless specific regulatory or statutory requirements are cited.  
The use of the word should in FDAs guidances means that something is suggested or 
recommended, but not required. 
 
 
II.  BACKGROUND 
 
FDA has become aware of potential biotin interference with IVDs that use biotin/avidin 
interactions as part of the device technology.  Many IVDs use biotin technology due to its ability 
to bond with specific proteins which can be measured to detect certain health conditions.  For 
example, biotin is used in hormone tests and tests for markers of cardiac health like troponin.  
Biotin, also known as vitamin B7, is a water-soluble vitamin often found in multi-vitamins, 
prenatal vitamins, and dietary supplements marketed for hair, skin, and nail growth (Ref. 1).  
Biotin levels in samples from patients who consume more than the recommended daily intake for 
biotin can cause falsely high or falsely low results, depending on the test principle.  Incorrect test 
results may lead to inappropriate patient management or misdiagnosis (Ref. 1).  FDAs concern 
regarding biotin interference was expressed in Safety Communications on November 28, 2017 
(Ref. 2) and November 5, 2019 (Ref. 3) and manufacturers of currently marketed devices have 
been working with FDA to address interference when it occurs.  Historically, devices using 
1  Contains Nonbinding Recommendations 
biotin/avidin technology have been assessed for biotin interference at the normal recommended 
daily doses of biotin (30 g per day, which results in plasma/serum biotin levels of < 1 ng/mL).  
However, several recent reports (Refs. 4-6) have described unanticipated biotin interference with 
the performance of some IVDs due to consumer use of dietary supplements that result in 
plasma/serum biotin levels of > 1 ng/mL.  In addition, extremely high biotin doses also have 
been observed (up to 300 mg per day, which results in plasma/serum biotin levels of > 1000 
ng/mL).   
 
This guidance describes FDAs recommendations for testing for biotin interference on devices 
that use biotin/avidin technology and communicating the results of such testing to the end-users, 
including via labeling.  The recommendations in the guidance are consistent with recent advice 
we have provided to individual manufacturers and sponsors that have consulted with the agency.  
They reflect FDAs thinking that labeling alone may not be sufficient to mitigate the risk of 
incorrect results from biotin interference in all cases.  Additional mitigation strategies may be 
considered when the risk of potentially incorrect results from biotin interference could 
significantly affect patient or public health. 
 
 
III.  RECOMMENDATIONS 
 
  Sponsors should contact the appropriate CBER or CDRH review division if biotin 
interference at clinically relevant analyte and biotin concentrations in patient samples is 
demonstrated. 
 
  We recommend that studies designed to test for biotin interference follow designs similar 
to those included in the most current version of Clinical Laboratory Standards Institute 
(CLSI) EP07, Interference Testing in Clinical Chemistry; Approved Guideline (Ref. 7). 
 
  Concentrations of biotin that reflect current trends in biotin consumption should be 
evaluated.  Biotin should be evaluated up to 3500 ng/mL, three times the maximum 
expected clinical concentration. 
The evaluation of biotin at this level is consistent with the recommendations in the CLSI 
standard and appropriate for minimizing the risk to patients from incorrect test results.   
 
  The samples should include analyte levels near the medical decision point(s) of the 
device. 
 
  For assays that are susceptible to biotin interference at concentrations less than 3500 
ng/mL, the concentration of biotin at which no interference is detected should be 
determined.  
 
  Information on biotin interference should be communicated to end users such as clinical 
laboratories and clinicians, including via the labeling1 of the device.  Relevant 
                                                 
1 Under 21 CFR 809.10(b)(10), the labeling accompanying an in vitro diagnostic product shall state known extrinsic 
factors or interfering substances affecting results.  More information regarding IVD labeling requirements is 
available at https://www.fda.gov/medical-devices/device-labeling/vitro-diagnostic-device-labeling-requirements. 
2  Contains Nonbinding Recommendations 
information to include in the labeling may be the percent difference or bias at each 
concentration tested for both qualitative and quantitative assays and the consequence of 
biotin interference (e.g., falsely elevated, falsely depressed), if observed.  
 
   
3  Contains Nonbinding Recommendations 
IV.  REFERENCES 
   
1.  Biotin:  Fact Sheet for Consumers, accessed March 20, 2019.  
https://ods.od.nih.gov/factsheets/Biotin-Consumer/ 
 
2.  FDA Safety Communication.  The FDA Warns that Biotin May Interfere with Lab Tests.  
November 2017.  https://www.fda.gov/medical-devices/safety-communications/fda-
warns-biotin-may-interfere-lab-tests-fda-safety-communication. 
 
3.  UPDATE:  The FDA Warns that Biotin May Interfere with Lab Tests:  FDA Safety 
Communication.  November 2019.  https://www.fda.gov/medical-devices/safety-
communications/update-fda-warns-biotin-may-interfere-lab-tests-fda-safety-
communication.  
 
4.  Piketty, M.L., et al. (2017) High-dose biotin therapy leading to false biochemical 
endocrine profiles:  validation of a simple method to overcome biotin interference.  Clin. 
Chem. Lab Med. 55(6):817825. 
 
5.  Chun, Kelly Y. (2017) Biotin Interference in Diagnostic Tests.  Clinical Chemistry 
63:619620.   
 
6.  Barbesino, G. (2016) The Unintended Consequences of Biotin Supplementation:  
Spurious Immunoassay Results Lead to Misdiagnoses.  Clinical Laboratory News, Bench 
Matters, December 13.  
 
7.  Clinical Laboratory Standards Institute. (2018) Interference Testing in Clinical 
Chemistry, 3rd Edition:  April.  https://clsi.org/standards/products/method-
evaluation/documents/ep07/ 
 
 
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