The Value Of Metals Testing For Risk . - Emerald Scientific

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The value of metals testing for riskmanagement and product safetyAnthony Smith, Ph.D, Chief Science Officer,OTCQB: EVIO#emeraldconference 20189 Emerald Conference

ABOUT EVIO LABSProvider of accredited analytical testing and industry research in7 different North American states & provincesSERVE CONSUMER DEMANDSAFETYHEALTHMAINTAIN HIGH STANDARDSCOMPLIANCEAND R&DTESTINGADVISORY CHEDUCATIONFORMULATIONSupport Health,Safety, EducationORELAP AccreditedCannabis Testing Labsin OR; ISO 17025 inCO, FL, CA, MA &Alberta, CAExpertise to SupportR&D, ProductFormulation, GrowOptimizationOTCQB: EVIO#emeraldconference 20189 Emerald Conference

Heavy Metal Contamination Standards & Limits:EU: EC Regulation 1881/2006 Maximum Levels for Contaminants in FoodstuffsAs 0.1-0.3 ppm, Cd 0.1-0.1 ppm, Pb 0.01-0.5 ppm, Hg 0.1-1.0 ppmUN: Codex Standard 193-1995 General Standard for Contaminants and Toxins inFood and FeedAs 0.1-0.5 ppm, Cd 0.2-0.9 ppm, Pb 0.1-0.3 ppm, Hg 0.1-1.6 ppmFDA: Has Set Standards for Some Foodstuffs, e.g. Shellfish, Children's Foods, FruitJuices and Bottled Water.#emeraldconference 20189 Emerald Conference

Heavy Metal Contamination:“Although the FDA has set specific levels for arsenic, lead, cadmium, and mercury in a variety of foods, more generally, the FDAhas—and uses—the authority to take action on a case-by-case basis where a particular food is found through routine or targetedtesting to be adulterated” --FDA Statement on Metals in FoodsAverage Inorganic Arsenic, Cadmium and Lead in CarrageenanProductAverage ArsenicAverage CadmiumAverage LeadNumber of SamplesCarrageenan181 ppb449 ppb251 ppb10Cadmium and Lead in Infant/Toddler FoodsProduct CategoryCereal - Infant/Toddler (rice)Cereal - Infant/Toddler (multigrain)Cereal - Infant/Toddler (non-rice)ApplesCereal - Oat RingGrapesJuice - GrapeJuice Boxes and PouchesPeanut ButterQuinoaRaisinsStage 2 Toddler FoodsTeething BiscuitsToddler PuffsAverage Cadmium(ppb)Range of Cadmium(ppb)Average 79.219.38.5 - 43.043.0 - 52.04.6 - 63.31.7 - 3.56.1 - 27.41.7 - 6.40.1 - 5.10.1 - 1.723.5 - 2350.4 - 61.01.6 - 5.11.7 - 31.11.7 - 33.44.2 - 1Range of LeadNumber of Samples(ppb)5.0 - 82.06.0 - 8.00.4 - 17.03.33.3 - 16.43.3 - 7.60.3 - 41.30.3 - 17.03.3 - 45.20.4 - 98.01.8 - 1511.0 - 22.22.0 - 1313.3 - 91.0766301030103040293023352731#emeraldconference 20189 Emerald Conference

FDA Sponsored Research:Mindak WR, et al. (2008) Lead in women's and children's vitamins.J Agric Food Chem. v. 56Number of SamplesHeavy Metal Contamination:ppmLead Content in Women's and Children's SupplementsProductFormAfter Baby Boost 1NutriBiotic Ultimate MatrixApex FIT Multivitamin Women'sRite Aid Prenatal TabletsEsteem Total WomanPreferred Plus Women's DailyUno Diario MujeresVegan PrenatalMaxi Teen SupremeA to Z Naturals Wow! PreNatalWomen's One DailyWhole Source Complete formula For WomenGenSpec VF-1 for African American FemalesGNC Women's Ultra Mega ActiveVitamin Source Prenatal CompleteGNC Women's Prenatal with IronPerry PrenatalKal Enhanced Energy Teen et/CapsuleLead stimated .9425.521.211.813.41.874.731.630.6671.78Population Assessment GroupPregnant and lactating womenAdult womenAdult womenPregnant and lactating womenAdult womenAdult womenAdult womenPregnant and lactating womenOlder children (7 years)Pregnant and lactating womenAdult womenAdult womenAdult womenAdult womenPregnant and lactating womenPregnant and lactating womenPregnant and lactating womenOlder children (7 years)#emeraldconference 20189 Emerald Conference

Heavy Metal Contamination:Best resource for “typical” heavy metal content in market foods:#emeraldconference 20189 Emerald Conference

Heavy Metal Contamination:Enough about food and supplements; What about smoked or inhaled products?Are there manufacturing control limits or regular testing of tobacco and cigarettes?Reported metals in US cigarettes:ArsenicCadmiumChromiumNickelLead0 - 0.24 ppm0.46 - 6.48 ppm0.06 - 1.11 ppm0.07 - 1.19 ppm0 - 1.58 ppmMetal conc.Cigarette wt.Mean transfer rateX Cigarettes per dayEstimated Daily Exposure“Median estimated availabilities of As, Cd, and Cr exceedthe ‘no significant risk levels’ (NSRL) as defined by theState of California all participants’ estimated exposuresto Cd would exceed the NSRL, while 50% would exceedthe NSRL for As. The estimated potential exposure to Pbis approximately 1/3 of the maximum daily dose (MADL)”“The average metal concentrations observed per gram of unburned tobacco were comparable to amounts found in cigarettes on the US,Canadian, and Brazilian markets, but less than those found in cigarettes purchased in China”Caruso RV, et al. (2014) Int J Environ Res Public Health. 11(1)Ashraf MW (2012) Sci World J. ID 729430Bailey EJ, et al. (1957) Br J Cancer. 11(1)Panta YM, et al. (2008) J Analysis and Applied Pyrolysis. 83(1)Reported metal content in cigarettes and cigars in AsiaArsenic20 – 50 ppmCadmium0.9 – 2.8 ppmLead1.6 – 3.6 ppmMercury0.02 – 0.1 ppm#emeraldconference 20189 Emerald Conference

Heavy Metal Contamination:Are there manufacturing control limits or regular testing of tobacco and cigarettes?Reporting Harmful and Potentially HarmfulConstituents in Tobacco Products andTobacco Smoke Under Section 904(a)(3) ofthe Federal Food, Drug, and Cosmetic Act“FDA recognizes that industry may be unable to meet the deadlinedue to current testing limitations. In recognition of this, the draftguidance identifies 20 HPHCs from the full list of 93 HPHCs, forwhich testing methods are well established and widely available.FDA is currently exercising enforcement discretion to requirereporting about these 20 constituents.”Abbreviated List of Harmful and Potentially Harmful ConstituentsHPHCs in CigaretteSmokeHPHCs in SmokelessTobaccoHPHCs in -ButadieneCarbon NNKNNNToluene#emeraldconference 20189 Emerald Conference

M e ta lsA c tio nC A L O QL im itsF o rD a ta in T h isS tu d yF L L O QC A IN HA r s e n icC A O th e r & F LC A IN HC a d m iu mC A O th e r & F LC A IN HLeadC A O th e r & F LC A IN HM e rc u ryC A O th e r & F L0 .00 .5P P M(µ g g-123)#emeraldconference 20189 Emerald Conference

Instrument and Method Slide: Shimadzu ICPMS-2030 SystemsSample preparation– Microwave digestion in Aqua RegiaCalibration and QCs Blank – Consisting only of the reagents used for digestion and dilution. Fortified Blank – Identical to the blank sample, but spiked with 5 ppb As, Cd, Hg, and Pb. Matrix – Reagents used for digestion and dilution, along with 0.5 g cannabis flower Fortified Matrix – Identical to the matrix sample, but spiked with 5 ppb As, Cd, Hg, and Pb.Validation and method performance (FL and CA)#emeraldconference 20189 Emerald Conference

Heavy Metals in Florida Cannabis: Over 7100 unique samples tested (October 2017 – February 2019)Cannabis flower, Concentrates & Extracts and CIPsISO 17025, validated method (Shimadzu ICP-MS with microwave digestion)Study statistics:Analyte LOD (ppm) LOQ (ppm) Action Limit (ppm) # above LOD # above LOQ % above LOQ # 1592.2%69Cadmium 9%56Fail %0.0%1.0%0.5%0.8%#emeraldconference 20189 Emerald Conference

Heavy Metals in Florida Cannabis: Over 7100 unique samples tested (October 2017 – February 2019)Cannabis flower, Concentrates & Extracts and CIPsISO 17025, validated method (Shimadzu ICP-MS with microwave digestion)M e ta lsA b o ve L O Q(0 .2 P P M ) inF lo w e r a n dC o n c e n tra te&Figure 1: Heavy Metals Detected Above LOQ (0.2vs Concentrate & Extract SamplesE xppm)t r a cint CannabisS a m p l FloweresA r s e n i c ( F lo r )A r s e n ic (C & E )C a d m iu mC a d m iu m( F lo r )(C & E )L e a d ( F lo r )L e a d (C & E )M e r c u r y ( F lo r )M e r c u r y (C & E )A b o ve0123456P P M7(µ g g8-191040C a l.6080 100)#emeraldconference 20189 Emerald Conference

Heavy Metals in Florida Cannabis: Over 7100 unique samples tested (October 2017 – February 2019)Cannabis flower, Concentrates & Extracts and CIPsISO 17025, validated method (Shimadzu ICP-MS with microwave digestion)Data Trend Analysistypical Failed sample has more than one target detected above LOQValue to CustomersValidating components, ingredients, processesStory slides1. Tracking down Cadmium in two producers operations2. Lead contamination from process hardware#emeraldconference 20189 Emerald Conference

Heavy Metals in Florida Cannabis: Over 7100 unique samples tested (October 2017 – February 2019)Cannabis flower, Concentrates & Extracts and CIPsISO 17025, validated method (Shimadzu ICP-MS with microwave digestion)production methodButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal FatButter/Animal rateConcentrateConcentrateCrystalline talline rsenicResult .781.671.65#emeraldconference 20189 Emerald Conference

Isolating ‘industrial contamination’ in clients agprocesses Prelimiary routine sampling and testing of early flower development showed intermittent ‘streaks’ ofCd in their greenhousesData .4951.39ND1.07#emeraldconference 20189 Emerald Conference

Isolating ‘industrial contamination’ in clients agprocesses Prelimiary routine sampling and testing of early flower development showed intermittent ‘streaks’ ofCd in their greenhousesData .4951.39ND1.07aerial partsCadmiumND - 0.5 ppmrockwool bitsCadmium5.3 - 80.2 ppmroot tissueCadmium0.02 – 0.08 ppmsoilCadmiumNDother p.o.e, e.g. water, foliar spray,. etc. ND#emeraldconference 20189 Emerald Conference

Screening vape cartridges and filling equipment Sample prep scheme (1 cart, opened and rinsed with aqueous 20% Nitric Acid)Range of dilutions used for scaleQC’s: Blank leachate and leachate solution spiked w/ standardsPb (ng/mL)SampleBlank leachateNDLCS 6E1360.0046 253.0047 mean262.6LCS recovery 92.3%Descr.QCQCV1V1V1V1V1Z CellZ CellZ CellZ CellZ Cell#emeraldconference 20189 Emerald Conference

Scientific Conclusions:Technical The main technical barrier is risk for sample-to-sample carryover and maintaining cleanblanksIn California, acid microwave digestion of ½ gram samples may incur increased safety riskPractical The gathering of large heavy-metals data sets for a particular commercial commodity isunprecedented and has epidemiological implicationsIt is easy to contaminate a cannabis product without having heavy metals in your soilRisk management, process control and surveillance can keep heavy metals out of your product#emeraldconference 20189 Emerald Conference

#emeraldconference 20189 Emerald Conference

Rite Aid Prenatal Tablets Tablet/Capsule 1.13 1.31 Pregnant and lactating women Esteem Total Woman Tablet/Capsule 0.861 3.61 Adult women Preferred Plus Women's Daily Tablet/Capsule 0.749 1.31 Adult women Uno Diario Mujeres Tablet/Capsule 0.65 1.05 Adult women Vegan Prenatal Tablet/Capsule 0.63 1.03 Pregnant and lactating women

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