STAKEHOLDER PANEL ON DIETARY SUPPLEMENTSDIETARY SUPPLEMENTS

Background & Fitness for Purpose

TURMERIC

Chair: AnikoM Solyom GAAS AnalyticalChair: Aniko M. Solyom, GAAS Analytical

Westin Bonaventure Hotel, Los Angeles, California

September 25, 2015

SPDS Advisory Panel Priorities (Set 4)

Factors:Factors: • Research interests • economic importance• availability of methods & standards 

Scopes: h d f dPrimary: Quantitative methods for curcuminoids

Secondary: Reduced forms 

Turmeric (Curcuma longa L.)Common names: turmeric,  turmeric root, Indian saffronMember of the ginger family, Zingiberaceae

Turmeric rhizoma

UsesCulinary:

fl i d l i t• flavoring and coloring agent• main spice in curry

Traditional Chinese and Ayurvedic medicine:• topical application for eczema and wound healing• aid digestion and liver function• relieve arthritis pain• regulate menstruation

Source: NCCIH Dietary Supplement Database (https://nccih.nih.gov/health/turmeric/ataglance.html

• regulate menstruation

Current research:• osteoarthritis, Alzheimer disease, eye inflammation, • colorectal cancer, Crohn’s disease, diabetes, stomach upset• gingivitis, stomach ulcer, irritable bowel syndrome, RA and more… 

Turmeric (Curcuma longa L.)

Spectra of turmeric extract

Approx. 5% of the plant is curcumin

CurcuminoidsO OO O

CH3O

OH

CH3O

OH

O O

OH

CH3O

OH

Curcumin

MW:368

Demethoxycurcumin

MW:338OH OH

O O

OH OH

MW:338

Bisdemethoxycurcumin

MW:308

Significance 

• In the Dietary Supplement Database (DSLD) 1,044 products contained turmeric and/or curcumin(oids) and/or extracts                     (out of total 42,000 DS)

f h d /• 47% of these products turmeric/curcumin as a component of a blend

Source: Leila G. Saldanha, PhD, RD, Office of Dietary Supplement, NIH. Personal communication

Significance190 clinical trials190 clinical trials 

between 

1996 and 2015

(http://clinicaltrials.gov)

Challenges• Nomenclature:• Nomenclature:

– Turmeric, turmeric oil

– Curcumin, curcuminoids

– Standardized to x% curcumin

AdulterationI di i d i i f• Indian turmeric trade types curcumin contents ranging from 2.1% to 8.6%, with an average of 4.8%. 

• Curcuma longa L.  adulterated with wild species: Curcuma zeodaria, Curcuma malabarica – toxicity and poor quality

• Adulterated with artificial colors – metanyl yellow 

• Saffron is adulterated with turmeric

Challenges

• Clinical Phase I studies have shown that the blood serum levels of curcumin are in the ng/mL range after oral doses of up to 8 g of curcumin, suggesting very low gastro‐intestinal bioavailability

• The reasons for the low oral bioavailability of curcumin are not yet knowny– chemical instability (degradation products are vanilin, ferulic acid, 

feruroyl methane) 

– rapid metabolism

– poor absorption

– accumulation in cells of the gastro‐intestinal tract

Analytical NeedsQ i i h d f i id i• Quantitative method for curcuminoids in– Raw material (plant material without authentication)

– Extracts

– Finished products containing only turmeric and/or curcuminoids

– Finished products containing other ingredients (vitamins, other DS, herbs, metanyl yellow)

– Biological fluids (?)Biological fluids (?)

• Quantitative method for curcuminoids in– Capsules 

– Tablets

– Tinctures

– Softgel capsules

Existing Methods

S iFi d h “ i d lid i ” d “2014 2015”• SciFinder search: “turmeric and validation” and “2014‐2015” yielded 97 references

• Spectrophotometric method for  the estimation of curcumin in bulk and pharmaceutical formulation

• 1H‐NMR and PCR for detecting Curcuma longa wild species adulterantsadulterants

• HPLC and LC/MS are widely used analytical techniques

History of turmeric method validation

• February 16, 2010: AOAC Expert Review Panel 

• 2009: “Call for Methods” to quantify curcuminoidcontent in dietary supplements and raw materials.

• 38 methods was submitted

evaluates the submitted methods and selects TUR‐36, developed by GAAS Analytical  as the method recommended for further consideration.

History of turmeric method validation

• March 22 2012: NIH publishes a Request for Quotes

• October 29, 2012: ChromaDex announces the award f t th d d l t lid ti t t

• March 22, 2012: NIH publishes a Request for Quotes (RFQ) for the “Method Optimization and Single‐Laboratory Validation of an Analytical Method: Total Curcuminoid Content of Turmeric” 

of two method development validation contracts. “These contracts will create single laboratory validated methods for the quantitative determination ….. of total curcuminoid content in turmeric.”

Fitness for Purpose (2010)

Turmeric Method Fitness For Purpose Statement (2010)“The method will be able to quantify total curcuminoid content, calculated as the sum of curcumin, demethoxycurcumin, and bis-demethyoxycurcumin, in turmeric [Curcuma longa Linn.] powdered botanical raw materials, extracts, and dietary supplement finished products containing turmeric extract, alone or in combination with other herbs such as Piper nigrum and Boswellia serrata. The method must be able to quantify curcuminoid content at levels up to 95% w/w in test materials. The lower limit of quantitation will be determined by the Expert Review Panel.”

Fitness for Purpose (proposal)

Questions/notes to the stakeholders and member of the working group:

• What happened between 2009 and 2015?

– More complex finished products on the market, l l h b dcontaining multiple herbs and vitamins

– Number of clinical trials increased – need of analytical support

SPDS Advisory Panel Priorities (Set 4)

Factors:Factors: • Research interests • economic importance

• availability of methods & standards 

Scopes: Primary: Quantitative methods for curcuminoids

Secondary: Reduced forms 

Challenges

• Clinical Phase I studies have shown that the blood serum levels of curcumin are in the ng/mL range after oral doses of up to 8 g of curcumin, suggesting very low gastro‐intestinal bioavailability

• The reasons for the low oral bioavailability of curcumin are not yet knowny– chemical instability (degradation products are vanilin, ferulic acid, 

feruroyl methane) 

– rapid metabolism

– poor absorption

– accumulation in cells of the gastro‐intestinal tract

Metabolism of curcumin in Caco‐2 cellsOH OOH O

H3CO

HO

OCH3

OHCUR

OH O

H3CO

HO

OCH3

OHhexahydro-CUR

CUR glucuronide

CUR sulfate

hexahydro-CUR glucuronide

hexahydro-CUR sulfatehexahydro CUR

OH OH

H3CO

HO

OCH3

OHoctahydro-CUR

octahydro-CUR glucuronide

octahydro-CUR sulfate

Analytical Needs• Quantitation of parent compound(s) and metabolites inQuantitation of parent compound(s) and metabolites in 

biological fluids to support clinical trials

blood

urine

• Reduced forms in dietary supplement finished products? Probably too expensive.

feces

Challenges

b l b l ll f• Low bioavailability ‐ very small amount of analyte

• Expensive instrumentation

• Radioactive tracing (?)

A il bilit f t d d• Availability of standards 

• Price of the standards

Available Metabolite Standards

• Dihydrocurcumin:– Used to be available from Indofine. Excerpt from a recent e‐mail: “Unfortunately I 

have bad news. We are unable to isolate dihydrocurcumin at this time. We made several attempts and were unsuccessful. For that reason we have to cancel this order. We are continuing to try, so I will keep you informed of our progress”

• Tetrahydrocurcumin (CAS 36062‐04‐1):– Santa Cruz Biotechnology 1 g $240

C t Ch i l C 1 $221 50– Crescent Chemical Company 1 g $221.50

– TLC PharmaChem, Cayman Chemical 10 mg $200

– Toronto Research Chemicals 100 mg $100, 1 g $120

• Hexahydrocurcumin (CAS 36062‐05‐2)– Crescent Chemical Company 10 mg $526

Fitness for Purpose (proposal)

Need clarification from the stakeholders and advisory board members

QUESTIONS??