Standardization and Quality Control Standardization and Quality Control in the in the

Mushroom Nutriceutical IndustryMushroom Nutriceutical Industry

Shu-ting ChangShu-ting Chang11 and and John A. BuswellJohn A. Buswell22

11Department of Biology, The Chinese University of Hong Kong; Department of Biology, The Chinese University of Hong Kong; 22Edible Fungi Institute, Shanghai Academy of Agricultural SciencesEdible Fungi Institute, Shanghai Academy of Agricultural Sciences

What are Mushroom Nutriceuticals?What are Mushroom Nutriceuticals?

‘Refined/partially refined/unrefined mushroom preparations derived from fruit

bodies, fungal mycelium or the spent culture fluid following mycelium growth in

submerged culture that possess nutritional and/or health-promoting properties and

which are consumed in the form of capsules or tablets as a dietary supplement’

(Chang & Buswell, World J. Microbiol. Biotechnol., 12, 473-476, 1996)

Market value of medicinal Market value of medicinal mushroomsmushrooms

1991 - US$1.2 billion (Chang, 1993)

1994 - US$3.6 billion (Chang, 1996)

1999 - US$6.0 billion (Wasser et al. 2000)

Current estimate – US$14 billion

Sales patternsSales patternsPre-1995: 99% of all sales of medicinal mushrooms and

their derivatives were concluded in Asia and Europe

<0.1% in North America

Recent years: both North and South American demands have

increased 20-40% annually depending upon the species

China is the major producer and China is the major producer and consumer of medicinal mushroomsconsumer of medicinal mushrooms

In 2000, >100 research units/institutes were engaged on R & D of medicinal mushrooms

Some 30-40 varieties of mushroom products for use as nutriceuticals/herbal medicines were manufactured in more than 200 factories

About 700 mushroom-based health food products, including >90 brands of G. lucidum products, were registered and marketed

(Lin, 2000)

Feedstocks for mushroom productsFeedstocks for mushroom products

Chang & Miles (2004) reported that:~77% of all medicinal mushroom products were

derived from fruiting bodies, which have either been cultivated commercially or collected from the wild

Only ~21% of all products were derived from fungal mycelia

Approximately 2% were derived from culture filtrates

Mycelial- and culture broth-based Mycelial- and culture broth-based products increasing in importanceproducts increasing in importance

Demands for increased safety and quality control, and for year-round production

Submerged culture processes can readily be standardized under controlled conditions

Downstream processing of active components released into the culture medium usually involve relatively simple procedures

Production methodsProduction methods

Powdered whole wild/cultivated fruit bodies, primordia or mycelium

Hot water/EtOH/MeOH of extracts of wild/cultivated fruit bodies or mycelium

Extracts of powdered fruit bodies obtained using supercritical fluid CO2

Various combinations of the above

LABELLING

Of 40 mushroom products examined in shops/companies, only two (both Ganoderma products) had labelling that revealed the product was standardized to contain polysaccharides (12.5%) and triterpenes (4.5%)(S.T. Chang, unpublished)

More commonly, labels claimed only that each capsule

contained, for example, only pure natural lingzhi (Ganoderma lucidum) or a mixture of lingzhi and maitake (Grifola frondosa). The labelling occasionally claimed that the product contained no preservatives, artificial colouring or flavouring, and that it was manufactured on ‘licensed premises’. No nutritional information was provided and no other ingredients were listed. Although some companies provided validation reports relating to the polysaccharide content, as well as HPLC and HPTLC validation profiles, of powdered Ganoderma fruiting bodies and mycelium, these companies do not disclose the test data on the bottle/packaging containing their products

Comparison of triterpene and polysaccharide Comparison of triterpene and polysaccharide

contents of 11 commercial lingzhi productscontents of 11 commercial lingzhi products Nature of product Triterpenes (%) Polysaccharide (%)

A - Fruit body extract 1.36 4.48

B - Fruit body extract 2.36 5.32

C - Fruit body extract 1.88 15.70

D - Fruit body extract 1.06 10.97

E - Fruit body extract 0.44 7.51

F - Fruit body extract 1.78 6.18

G - Fruit body extract 1.44 13.30

H - Fruit body extract 0.50 15.80

I - Fruit body extract 7.82 7.66

J - Fruit body powder 0.46 1.10

K - Mycelium powder Undetectable 12.78

Nature of MushroomNature of Mushroom Nutriceuticals Nutriceuticals

Some, e.g. lentinan, is sold in refined or purified form

Overwhelming majority of mushroom-based nutriceutical products currently available are not single compounds but combinations of several individual components that together contribute to the overall bioactivity of the product

Therefore, it is important that the future development of mushroom nutriceuticals should not be focused solely on the isolation and bioactivity of individual mushroom components, otherwise synergistic effects will be overlooked

Current regulations relating to Current regulations relating to Mushroom NutriceuticalsMushroom Nutriceuticals

Position is problematic due to:

1. the considerable variation in the regulations applicable to the different countries active in the marketplace

2. the borderline position occupied by mushroom nutriceuticals between food and medicine

Bagchi D. 2008. Nutraceutical and functional food regulations in the United States and around the world. Academic Press, New York, 462pp.

Proposed Protocol for Obtaining Proposed Protocol for Obtaining Quality Mushroom ProductsQuality Mushroom Products

FIVE “G” guidelines

- originally proposed for adoption as a basis for the manufacture of quality mushroom products from mushroom fruit bodies

(Miles and Chang 1997, Chang 2006)

GLP (Good Laboratory Practice)

GAP (Good Agricultural Practice)

GMP (Good Manufacturing Practice)

GPP (Good Post-formulation Practice)

GCP (Good Clinical Practice)

GLP (Good Laboratory Practice)GLP (Good Laboratory Practice)

Source and nature of the mushroom strain must be clearly documented

Strains should be properly maintained and preserved without contamination or degeneration

GAP (Good Agricultural Practice)GAP (Good Agricultural Practice) High priority given the propensity of mushrooms to

accumulate potentially harmful contaminants from the growth environment

Strictly defined and maintained growth and harvesting conditions for commercially cultivated fruit bodies

Growth substrate and ancillary material (e.g. casing) should be free of harmful contaminants – e.g. heavy metals

Stipulation of the physical growth parameters (e.g.

temperature, relative humidity, illumination regimes)

Good sanitary growth conditions should prevail (e.g. free from contaminated water and polluted air, microbial contamination, insect infestation)

GAP (Good Agricultural Practice) - IIGAP (Good Agricultural Practice) - II

GAP is important not only for the quality and safety of the product - it can also affect the yield of the desired bioactive component

Log-grown xianggu (L. edodes) contained more high-molecular weight polysaccharides (HMWP) than sawdust-grown mushrooms and, among the log-grown shiitake, both mushroom strain and tree species influenced HMWP content

GMP (Good Manufacturing Practice) GMP (Good Manufacturing Practice) Development, standardization and constant monitoring

of contamination-free downstream processing protocols and parameters (e.g. pre-treatments prior to comminution, comminution methods, extraction temperatures, extraction times, solvents)

Although most mushroom nutriceuticals are heterogeneous in nature, levels of the main active constituents of a particular product should, as far as the nature of that product allows, be determined and disclosed in order to guarantee quality, authenticity and dosage formulation - e.g. content of the major triterpenoid and/or polysaccharide components for Ganoderma products

GMP (Good Manufacturing GMP (Good Manufacturing Practice) - IIPractice) - II

Establishment of certified testing centres in order to provide product validation/information to manufacturers, retailers and consumers

GPP (Good Post-formulation Practice)GPP (Good Post-formulation Practice)Chemical and microbiological analyses to ensure

that all types and levels of chemical (e.g. heavy metals) and microbiological contamination fall within safe limits

Determination of optimum storage conditions

and rates of inactivation/deterioration over time relating to the major active ingredients of marketed products in order to ascertain shelf life and to establish appropriate ‘sell-by dates’ nb: exoglucanase-mediated degradation of lentinan during storage of L. edodes fruit bodies

GCP (Good Clinical GCP (Good Clinical Practice)Practice)

High quality clinical trials, including double-blind

studies, should be conducted over the longer term:

1. to confirm claims of product bioactivity

2. to facilitate product formulation

3. to determine an appropriate dosage level for

an effective health-promoting

outcome

Concluding RemarksConcluding Remarks

1. There is growing experimentally-based evidence to suggest that dietary supplements based on bioactive compounds extracted from mushrooms (mushroom nutriceuticals) increase resistance to disease and, in some cases, causes regression of a diseased state

2. In many cases, these products appear

to enhance the host immune response

Concluding Remarks - IIConcluding Remarks - II

3.3. They have extraordinary low toxicity, even They have extraordinary low toxicity, even at high doses, and are apparently lacking in at high doses, and are apparently lacking in various side effects that frequently accompany various side effects that frequently accompany the use of synthetic drugs the use of synthetic drugs

4.4. Since mushrooms have a long tradition as a Since mushrooms have a long tradition as a food source, many mushroom feedstocks are food source, many mushroom feedstocks are categorised as ‘generally considered safe’categorised as ‘generally considered safe’

Concluding Remarks - IIIConcluding Remarks - III

5. Increasing interest in mushroom nutriceuticals is likely to continue worldwide in view of the challenges and opportunities they represent, and their economic value may ultimately surpass that of mushrooms currently produced for food

6. Consumers nowadays are more demanding and better informed. Therefore, it is crucial that mushroom products be of high, reproducible quality and free from potentially harmful substances in order to earn the enduring public credibility essential for future market expansion

Concluding Remarks - IVConcluding Remarks - IV7. Widely accepted procedures relating to

feedstock production, downstream processing, product safety and stability, and product efficacy need to be developed and continually improved

8. Introduction of a registration system for mushroom nutriceuticals based on information obtained using these procedures would achieve the highest level of quality assurance, and provide reputable manufacturers with an effective marketing strategy while helping to eliminate less reliable manufacturers/traders

Concluding Remarks - VConcluding Remarks - V

9. Accurate disclosure of this information to the consumer will create an automatic monitoring system, and contribute enormously to the overall integrity of the mushroom nutriceutical industry

Thank youThank you