Guidance on Safety assessment of botanicals and botanical ...
Botanical Integrity: Significance of Fit for Purpose Tests During … · 2018-05-15 · WHY...
Transcript of Botanical Integrity: Significance of Fit for Purpose Tests During … · 2018-05-15 · WHY...
Amit Chandra, Ph.D
AMWAY R&D
Phytochemical Fingerprint
Botanical Integrity: Significance of Fit for Purpose Tests During
Botanical Product Development Process
WHY BOTANICALS IN SUPPLEMENTS:
Colorful fruits and vegetables are healthy sources of beneficial nutrients.
WHO recommends five servings (about 450g) of fruits and vegetables every day to meet these requirements.
75% of adults fail to meet this recommendation.
Supplements containing vitamins, minerals and botanicals help to close this gap.
Hence “quality of” and “choice of” right botanical for the right health benefit is extremely important !
(Nutrilite Health Institute-Global Phytonutrient Report,
British J. Nutrn, 2014)
TYPICAL FORM OF BOTANICAL INGREDIENTS, ACTIVES v/s MARKERS:
STANDARDIZED EXTRACT: Ingredient with certified / consistent levels of
phytonutrients produced via controlled manufacturing / extraction process
MARKERS (are typically quality indicating): Phytonutrients that are “unique to” and used as analytes for
identifying and quantifying the botanical ingredient.
Please note: It is typically the whole ingredient that offers / maintains health benefit (in a supplement)
not specific phytonutrients !
ADULTERATION OF BOTANICALS IN DIETARY SUPPLEMENT COMPROMISING ITS INTEGRITY:
(1)If any valuable constituent has been in whole or in part omitted or abstracted therefrom; or (2) if any substance has been substituted wholly or in part therefor; or (3) if damage or inferiority has been concealed in any manner; or (4) if any substance has been added thereto or mixed or packed therewith so as to increase its bulk or weight, or reduce its quality or strength, or make it appear better or of greater value than it is.
21 US Code 342 – Adulterated Food
TYPICALLY OBSERVED
ADULTERATIONS IN BOTANICALS
ECONOMICALLY MOTIVATED ADULTERATION (Intentional): Fortifying botanical ingredients by adding “markers” to pass quantitative tests !
It is by design, not accident ! Markers can be from synthetic / natural source
ACCIDENTAL ADULTERATION (Incidental): Due to carelessness in chain of custody
No traceability ! supply chain, violation of cGMP, cross contamination etc.
FORTIFYING OR REPLACING BOTANICALS WITH PRESCRIPTION DRUGS (Intentional)
For quick results ! In weight loss, body building, sexual enhancement
A holistic / qualitative analytical approach to identify the correct / intended botanical source SHOULD ALWAYS PRECEED its quantification in the product.
HENCE, THE CHOICE OF FIT FOR PURPOSE ANALYTICAL METHOD IS OF PRIME IMPORTANCE HERE
Chandra, A., in Korea-China Joint International Phytonutrient Symposium, Seoul, 2016, 147-156.
What is the purpose of method OR what is the question that is being asked (ID or Quant) ?
Is there some benchmark sample (authentic sample/control/reference) for comparison ?
Nature of botanical INGREDIENT (dehydrate or extract) ?
ID of whole plant or a specific plant part / processed ingredient ?
QUANTIFICATION OF INGREDIENT ? Genus spp and / or phytonutrients (markers) ….?
ANSWERS TO ABOVE WILL HELP TO CHOOSE THE PROPER ANALYTICAL TEST METHOD
AKA “FIT FOR PURPOSE”
CRITERIA TO CHOOSE FIT FOR PURPOSE METHODS?
AUTHENTIC BOTANICAL SAMPLE / CONTROLS / REFERENCE
• CERTIFIED REFERENCE MATERIALS
• VOUCHERED SPECIMENS,
• SELF COLLECTED / GENERATED BASED ON HISTORICAL USE, (with
documentation)
• ETC…..
DEPENDS ON YOUR PURPOSE.
SAMPLE MUST REPRESENT MATERIAL OF INTEREST
Example: Matrix, Harvest time, specific plant part, Standardization level
IT HAS TO
BE APPLES
TO APPLES
COMPARISON
QUALITATIVE METHODS USED FOR BOTANICAL ID
GENETIC: Full genomic sequence
DNA barcoding
Next generation sequencing (NGS)
BIOLOGICAL: Micro and Macroscopic, organoleptic…
CHEMICAL: (Spectroscopic) Patterns, Phytochemical Fingerprints
(Qualitative, Holistic)
Selected method must be
appropriate (fit for purpose) for
answering the question that is
being asked!
Khan, I.A., Smillie, T. J.Nat. Prod., 2012, 75 (9), 1665-1673.Betz, J.M., et al. Anal. Bioanal. Chem., 2007, 389, 19-25.
TIPS FOR FLAGGING BOTANICALS THAT ARE
VULNERABLE FOR THEIR INTEGRITY (PRONE TO
ADULTERATION)
• RESTRICTED / LIMITED SUPPLY CHAIN
• COMPLEX STANDARDIZATION PROCESS
• HIGH COST
• GENERIC PHYTONUTRIENT MARKERS
• ONE GENUS, MULTIPLE SPECIES
• LACK OF MANF. – SUPPLIER PARTNERSHIP
IN THIS CONTEXT,
I WILL NOW SHARE WITH YOU
TYPICAL EXAMPLES OF BOTANICAL SUPPLEMENTS IN MARKET THAT ARE VULNERABLE FOR ADULTERATION !
SIGNIFICANT INCREASE IN SUPPLY CHAIN
LOWERING OF COST
GENERIC PHYTONUTRIENT TARGET FOR STANDARDIZATION (Total flavonol-glycosides)
EXAMPLE 1: GINKGO BILOBA LEAF EXTRACT
Chandra A; Li Y; Rana J; Persons K; Hyun C; Shen S; Mulder T., J. Func. Foods, 3, 2011, 107-114
FIT FOR PURPOSE METHOD: PHYTOCHEMICAL FINGERPRINT
OF FLAVONOL GLYCOSIDES BEFORE HYDROLYSIS
RQ, K and I
This is the signature profile / fingerprint of
“authentic ginkgo leaf extract 24/6”
RESULTS AND CONCLUSIONS
All
Passed
24%
spec
Chandra A; Li Y; Rana J; Persons K; Hyun C; Shen S; Mulder T., J. Func. Foods, 3, 2011, 107-114
FLAGS:
SIGNIFICANT INCREASE IN SUPPLY CHAIN
LOWERING OF COST
GENERIC STANDARDIZATION (Total Anthocyanins)
ONE GENUS (Vaccinium spp) MANY SPECIES
EXAMPLE 2: BILBERRY (Vaccinium myrtillus)
Chandra A., Rana J., Li Y. J Agric Food Chem., 49 (8), 2001, 3515-3521
Li Y; Chandra A, Persons K, Stern N, Shen S. 127th AOAC International, 2013, Aug 25-28, Chicago IL, USA
FIT FOR PURPOSE TEST: HPLC Fingerprint of
anthocyanins in authentic Bilberry (as is )
This is the signature profile / fingerprint of anthocyanins in
“authentic bilberry extract”
Chandra A., Rana J., Li Y. J Agric Food Chem., 49 (8), 2001, 3515-3521
Looks like black rice
Looks like elderberry
Could be amaranth dye !
Li Y; Chandra A, Persons K, Stern N, Shen S. 127th AOAC International, 2013, Aug 25-28, Chicago IL, USA
RESULTS AND CONCLUSIONS
AUTHENTIC BILBERRY
BC BR BB1 BB2 GR BB BBn CG A
RECOMMENDED FIT FOR PURPOSE ANALYTICAL TESTS FOR ENSURING BOTANICAL INTEGRITY
DNA ORGANOLEPTIC
MACROSCOPIC
&
MICROSCOPIC
PHYTOCHEMICAL
WHOLE
DEHYD.
EXTRACT
STD. EXTRACT
SPECIFIC
PLANT PART * *
BLENDED
BOTANICALS
TEST
BO
TA
NIC
AL
*: RED FOR
EXTRACTS
* *
RED: Not fit for purpose
YELLOW: Limited fit
GREEN: Fit for purpose
RED FOR
UNKNOWN
BOTANICALS
Chandra, A., in Korea-China Joint International Phytonutrient
Symposium, Seoul, 2016, 147-156.
Quality By Design QbD ?
• Quality by Design (QbD) is a proactive and
systematic approach to development that begins
with predefined objectives and emphasizes
product and process understanding and process
control, based on sound science and quality risk
management.
WHY: FOR Advancement of botanical integrity,
safety and quality in the industry !
Khan, I.A., Smillie, T. J.Nat. Prod., 2012, 75 (9), 1665-1673.
Example of QbD for addressing botanical integrity (from
our latest market launch of Ayurvedic products in India)
Deployment of fit for purpose tests
(in that order):
1. Confirmation of correct genus and species by DNA
fingerprinting of whole plant
2. Verification of Non GMO status by DNA – PCR
3. Botanical ID (of specific plant part) by phytochemical
fingerprinting method using HP-TLC
4. Quantification of marker in extract and tablets by
chemical methods (HPLC) for claims, QA, stability etc.
5. Establishing quality standards with supply chain for each
production lot.
QbD, Right tests at the right stage-gates with full
documentation and partnership with supply chain and
compliance with regulatory = BOTANICAL INTEGRITY.
Example of QbD for Botanical
Integrity in development process
PRODUCT
DNA Fingerprinting on botanicals:
Where are we at ?
• DNA is the holy grail to address all problems !!!!! So much
confusion on the application of this technology in conjunction to
botanical integrity
• Industry trying to work to come to a harmonized approach for
recommending appropriate use of DNA test methods. AHPA-NIST
is leading an initiative in this direction with Industry
• FDA and USP do not recommend / endorse DNA testing as the only
test for genus and species. However, its use in conjunction to
relevant phytochemical methods is OK
• DNA is being legitimized as a test for botanical drugs in Korean
Pharmacopeia. However only at whole plant / herbal stage.
STEPS TO ENSURE INTEGRITY OF BOTANICALS
IN SUPPLEMENTS AND FOODS:
1. Full traceability of botanicals from farm to product
with audits and documentation
2. Active partnership with supply chain
3. Create and follow QbD process keeping end goal in
mind
4. Mandate use of fit for purpose analytical test
methods at all stage-gates during the development
process (seed to supplement)
5. Last but not the least, share – educate – publish –
talk – promote good practices !
ACKNOWLEDGEMENTS
Amway R&D
Amway-Nutrilite Botanical Authentication Team
Ole Miss NCNPR – Awesome Hosts
Industry Partners:
AHPA, AOAC, CRN, ABC, USP
….. and you the ELITE AUDIENCE !