Stefan Gafner, PhD

CSO, American Botanical Council

November 18, 2015

Oryx Hotel, Aqaba

BRAMA

Botanical Risk Assessment Training

Session VI

Introduction into adulteration

The American Botanical Council

•Non-profit educational organization

•Provides information on responsible and safe use of

medicinal herbs

•Members include consumers, healthcare

professionals, researchers, educators, industry

•Founded in 1988 with James A. Duke, Norman R.

Farnsworth and Mark Blumenthal as first Trustees of

ABC board

The ABC-AHP-NCNPR Botanical Adulterants

Program

•Program to educate herbal and dietary supplement industry

members about ingredient and product adulteration

• Initiated by three non-profit organizations:

• American Botanical Council (ABC)

• American Herbal Pharmacopoeia (AHP)

• National Center for Natural Products Research (NCNPR) at the

University of Mississippi

Adulteration

Adulteration is the action that results in accidental,

negligent, or intentional variations in identity, strength,

purity, and expected outcomes from a named or at least

implied identity of a herbal medicine, even if the standard

of identity was merely an organoleptic (sensory

observation) expectation

Adulteration of Botanical Ingredients

• Accidental: Substitution with/admixture of other

botanicals due to human error

• Economically-motivated: Substitution with/addition of

materials of substandard quality for financial gain

• Consequences: Adverse side effects, lack of efficacy, loss

of consumer confidence

Herbal DS Sales – Mainstream Multi-Outlet

Channel -- 2014

1. Horehound

2. Cranberry*

3. Echinacea

4. Black Cohosh*

5. Flaxseed and/or Oil

6. Valerian

7. Yohimbe**

8. Bioflavonoid Complex

9. Saw Palmetto*

10. Ginger***Subject to known spiking w/ Rx drugs

11. Aloe*

12. Milk Thistle*

13. Garlic

14. Cinnamon

15. Rhodiola*

16. Horny Goat Weed

17. Ginkgo*

18. Plant Sterols

19. Red Yeast Rice**

20. Elderberry*

Source: Symphony IRI & HerbalGram #107

*Subject to known or alleged/suspected adulteration

**Subject to known spiking w/ Rx drugs

Herbal DS Sales – Natural Channel -- 2014

1. Turmeric*2. Grass (wheat or barley)3. Flax seed and/or oil 4. Aloe*5. Spirulina6. Milk thistle*7. Elderberry*8. Maca*/**9. Echinacea*10. Oregano (oil & tincture)

Source: SPINS & HerbalGram #107

11. Saw Palmetto*12. Chia Seed and/or Oil13. Valerian 14. Garlic15. Echinacea-Goldenseal*16. Chlorophyll/Chlorella17. Cranberry*18. Ginkgo*19. Stevia20. Red Yeast Rice**

*Subject to known or suspected/alleged adulteration

**Subject to known spiking w/ Rx drugs

“Since the beginnings of civilization, once commerce develops, adulteration follows”

HerbalGram

Publications(As of September 2015)

Botanical Adulterant

Bulletins

• Arnica Flower

• Ashwagandha Root & Extract

• Bilberry Fruit Extract

• Black Cohosh Root & Rhizome

• Cranberry Fruit Extract

• Ginkgo Leaf Extract

• Ginseng, American & Asian Root & Extract

• Goldenseal Root & Rhizome

• Grapefruit Seed Extract

• Grape Seed Extract

• Maca Root

• Pomegranate Fruit & Extract

• Saw Palmetto

• Skullcap Herb

• Tea Tree Oil

• Vitex (Chaste Tree Berry)

Ginkgo biloba

Adulteration of Ginkgo biloba extracts

2003: Canada - 9 samples, 1 adulterated (11%)

2005: Canada - 14 samples, 4 likely adulterated (29%)

2006: China - 19 samples, 3 adulterated (16%)

2006: USA - 21 samples, 10 adulterated (48%)

2010: Germany - 10 samples, 7 likely adulterated (70%)

2011: USA - 8 samples, 3 adulterated (38%)

2012: Japan - 22 samples, 3 likely adulterated (14%)

2012: USA - 18 samples, 7 adulterated (39%)

2014: USA - 37 samples, no evidence of adulteration

2014: Australia - 8 samples, 3 adulterated (38%)

2015: UK – 30 samples, 12 adulterated (40%)

2015: USA - 25 samples, 11-19 adulterated (44-76%)

2015: Canada, in press - 14 samples, 11 adulterated (79%)

Ginkgo biloba Leaf Extract

Common Ginkgo Quality Concerns

• Commonly used adulterants of Ginkgo biloba:– Quercitin or kaempferol

– Rutin

– Flavone glycoside-rich extracts (e.g., Fagopyrum esculentum, Styphnolobiumjaponicum)

• These materials are relatively inexpensive & widely available.

• The addition of flavonoids enhance total flavonoid content of the extract.

• Adulterated products may go undetected, if

total flavone glycosides content is used as the

only QC criteria.

Asian Ginseng (Panax ginseng) Root Extract

Adulterated with Leaf Extract

• Companies are marketing Asian ginseng root extract

standardized to relatively high levels of ginsenosides at what

appear to be relatively low prices:

– 10-24%

• Analysis reveals:

– presence of chlorophyll in some samples

– uncharacteristic profiles of ginsenosides,

indicating presence of ginseng leaf extract

PomegranatePunica granatum, Punicaceae

http://blogs.knoxnews.com/knx/constantine/pomegranate.jpg

Pomegranate & Ellagic Acid

• Ellagic acid (EA) is an antioxidant compound

found in many fruits.

• Pomegranate extracts claim 40-70% EA.

• EA found in pomegranate: ca. 3%

– Up to 5% EA upon hydrolysis during extraction

• EA is inexpensive; made chemically and/or via

extraction from wood pulp.OH O O

O O

OH

OH

OH

Ellagic acid

Adulteration of Cranberry (Vaccinium macrocarpon)

Extract

• In the United States, cranberry is the third best-selling herb in the mass market and # 13 in natural channel with sales over $ 50 Mio*

• Beneficial effects for preventing urinary tract infection reportedly due to proanthocyanidins (PACs), in particular A-type

• Cranberries at 100 g fresh weight provide ca. 420 mg total flavan-3-ols, of which 56% (235 mg) are polymers**

• Other plants contain higher amounts of PACs and present cheaper sources

*Lindstrom A, Ooyen C, Lynch ME, Blumenthal M, Kawa K. HerbalGram. 2014;103:52-56.

** USDA database for proanthocyanidin content of selected foods. Beltsville (MD): Nutrient Data Laboratory, Agricultural Research Service, USDA 2004.

Adulteration of Cranberry (Vaccinium

macrocarpon) Extract

Mainly A-type PACs B-type PACs A-type and B-type PACs

Vaccinium macrocarpon Vitis vinifera Arachis hypogaea

Analysis of Commercial Cranberry Samples

• Fresh fruits (cranberries, blueberries, grapes, and raisins) and juices (cranberry, grape) purchased from markets1

• 10 commercial cranberry products obtained from manufacturer

• Samples analyzed by capillary zone electrophoresis and HPLC and subsequently clustered by statistical means (PCA)

• One cranberry extract aligned within grape cluster: absence of A-type PACs confirmed by high-resolution mass spectrometry

1Navarro M, Nunez O, Saurina J, Hernandez-Cassou S, Puignou L. J Agric Food Chem. 2014;62(5):1038-1046.

Procyanidin A2 Procyanidin B1

• Canadian cranberry extract manufacturer documented cranberry extract adulteration (likely with peanut skin extract) by HPLC-MS

Saw Palmetto Fruit / Berry ExtractSerenoa repens

Adulteration of Saw Palmetto

1Lindstrom A, Ooyen C, Lynch ME, Blumenthal M, Kawa K. HerbalGram. 2014;103:52-56.2Myers S. Nat Prod Ins. September 11, 20093Little DP, Jeanson ML. Scientific Reports 2013:3; DOI:doi:10.1038/srep03518

• Among 10 best-selling herb in mass market and natural channels

in the United States1

• Saw palmetto extracts often standardized to fatty acid content

• Palm, canola, olive, and coconut oils have been found as

adulterants in report from 20092

http://www.sfrc.ufl.edu/4h/Saw_palmetto/serrepws.jpg

• Occurrence tends to escalate

after a supply shortage

• Berries of close relative,

Acoelorrhaphe wrightii

reportedly sold as saw palmetto

based on DNA analysis3

Adulteration of

Arnica (Arnica montana) [a]

with

“Mexican Arnica” (Heterotheca inuloides) [b]

[both Asteraceae]

Walker KM, Applequist WL.

Adulteration of Selected Unprocessed

Botanicals in the U.S. Retail Herbal

Trade. Econ Bot, 2012.

Adulteration of

Thyme in Egyptian markets

• European Pharmacopoeia lists thyme (Thymus vulgaris) and

Spanish thyme (T. zygis) as acceptable species

• Most thyme traded as spice is a mixture of T. capitatus, T.

serpyllum, and T. vulgaris

• Statistical model with authentic samples was constructed using

UV spectrophotometry

Gad HA, El-Ahmady SH, Abou-Shoerb MI, Al-Azizia MM. A modern approach to the

authentication and quality assessment of thyme using UV spectroscopy and

chemometric analysis. Phytochem Anal. 2013;24(6):520-526.

• Commercial samples (n=12) collected

in Cairo and Aswan clustered around

T. vulgaris (9), Satureja montana (1)

or were of unknown identity (2)

Thymus vulgaris (Lamiaceae)

Adulteration of African mango seed Irvingia gabonensis (Irvingiaceae)

• Popularity in weight-loss

products.

• Currently no quality standards

• Methyl gallate, ellagic acid, &

derivatives as main

components.

• UHPLC/HRMS screening of 7

commercial materials indicated

adulteration, including regular

mango (Mangifera indica).

Sun J, Chen P. Ultra high-performance liquid chromatography with high-resolution mass

spectrometry analysis of African mango (Irvingia gabonensis) seeds, extract, and related dietary

supplements. J Agric Food Chem. 2012;60:8703−8709.

http://www.africanmangoguide.org/

#4

top-selling

herbal DS

in

mainstream

retail stores

in 2014

Black Cohosh Adulterants

Chinese cimicifuga • Sheng ma

Actaea dahurica

Xing an sheng ma

Actaea cimicifuga

Sheng ma

Actaea heracleifolia

Da san ye sheng ma

Black Cohosh AdulterationRecent Reports

2006: HPLC-ELSD (n=4): 25% adulterated1

2006: HPLC-MS (n=11): 36% adulterated2

2012: DNA barcoding (n=36): 25% adulterated3

2014: HPLC-MS/MS & DNA sequencing (n=25): 28% adulterated4

2015: FIMS, NMR & DNA (Sanger) sequencing5

Chinese Actaea raw materials (n=11): 85% adulterated

Finished products (n=14): authentication not possible by NMR, FIMS; DNA found in 5 samples: 20% adulterated

1He K, Pauli GF, Zheng B, et al., J Chromatogr A 2006;1112(1-2):241-2542Jiang B, Kronenberg F, Nuntanakorn P. et al., J Agric Food Chem 2006;54(9):3242-32533Baker DA, Stevenson DW, Little DP. J AOAC Int. 2012;95(4):1023-10344Masada-Atsumi S, Kumeta Y, Takahashi Y, et al., Biol. Pharm. Bull. 2014;37(3):454–460 5Harnly JM, Chen P, Sun J et al., Planta Med. 2015; in press

1He K, Pauli GF, Zheng B, et al., J Chromatogr A 2006;1112(1-2):241-2542Jiang B, Kronenberg F, Nuntanakorn P. et al., J Agric Food Chem 2006;54(9):3242-32533Baker DA, Stevenson DW, Little DP. J AOAC Int. 2012;95(4):1023-10344Masada-Atsumi S, Kumeta Y, Takahashi Y, et al., Biol. Pharm. Bull. 2014;37(3):454–460 5Harnly JM, Chen P, Sun J et al., Planta Med. 2015; in press

The botanical supply chain is global;

adulteration is a global challenge.

http://http://www.erau.edu