NOVEMBER 2021

TEST METHODOLOGY

Heavy Metals and Salmonella in Herbs and Spices
The purpose of this study was to determine the levelsof arsenic, cadmium, lead, mercury, and Salmonella
species in 15 popular and commonly consumed spices,and to assess any associated health risk(s). In addition,
we investigated possible adulteration, specifically theaddition of bulking and/or coloring agents, in four of
the 15 spices that are commonly adulterated in theseways. We tested five to 10 products each of basil, blackpepper, chili, coriander, cumin, curry powder, garlicpowder, ginger, oregano, paprika, saffron, sesame seed,thyme, turmeric, and white pepper. We tested two tothree samples of each product, for a total of about 330
samples. The products tested were selected based onsales data and data from shopper surveys of stores in
New York, New Jersey, and Connecticut. We purchased
the samples between October 2020 and January 2021
from stores and online from these three states. We
made an effort to include nationally available, private/store, and culturally relevant products, with each sample
representing a unique lot of each product.
SAMPLE PREPARATIONThe samples were either masked or transferred intobrown polyethylene jars, blind coded them to preserve
their identities, and shipped them overnight to two
independent, accredited laboratories for the analyses.
At the heavy metals lab, sample preparation or
homogenization was performed in fume hoods known
to be free from trace-metal contamination. At thesecond lab, microbiological (Salmonella) tests, botanicalidentification tests, and the test for coloring agentsor Sudan dyes were performed according to specified
methods. Water, sample containers, and other materials
used for the analyses were monitored for contamination,
where appropriate, to account for any biases in s
ample results.
TESTINGAnalysis for Arsenic, Cadmium, Lead, and Mercury
by Triple Quadrupole Inductively Coupled Plasma
Mass Spectrometry (IC-QQQ-MS), With Collision CellReaction (CRC)
The samples were prepared and analyzed in accordancewith the Association of Official Analytical Chemists(AOAC) Method 2015.01.
Arsenic Speciation Analysis by Ion ChromatographyInductively Coupled Plasma-Collision Reaction CellMass Spectrometry (IC-ICP-CRC-MS)
Select samples with high total arsenic results fromeach spice category were analyzed for total inorganic
arsenic and three organic arsenic species—monomethylarsonic acid (MMA), dimethyl arsinic acid (DMA), andtrimethylarsine oxide (TMAO).
Analysis for Salmonella
Samples were tested for the presence of Salmonella
following the Food and Drug Administration’s
Bacteriological Analytical Manual (FDA BAM) method(Chapter 5: Salmonella).
Botanical Identification Analyses
The identification of specific plant species in the tested
samples was conducted by deoxyribonucleic acid(DNA) extraction and polymerase chain reaction (PCR)
analysis, using specific primers for known bulking agentsor adulterants.
Analysis for Sudan Dyes
The samples were prepared and analyzed for seven
Sudan dyes by liquid chromatography tandem mass
spectrometry (LC-MS/MS) in accordance with theAmerican Spice Trade Association (ASTA) Method 28.
Sample analyses for chemical contaminants wereprecluded by at least a five-point calibration curve
spanning the entire concentration range of interest.
Calibration curves were performed at the beginningof each analytical day and verified during analysis.
The testing conformed to the quality control criteria
and performance requirements set in cited officialmethods, as well as to those under the lab’s
ISO 17025 accreditation.

CR.orgNOVEMBER 2021

TEST METHODOLOGYRISK ASSESSMENT
We estimated daily consumption of the spices using
the ¼ teaspoon or 0.5 grams per eating occasion FDA
Center for Food Safety and Applied Nutrition (CFSAN)
reference amount customarily consumed (RACC) ofspices and herbs, our test results, and the average bodyweights of adults and children 1 to under 6 years old.
We used the recommended body weights from theEnvironmental Protection Agency (2011 Exposure Factors
Handbook). For heavy metals test results below themethod detection limit (MDL), we used the method fromXue, et al. (2010), to estimate the average concentrationof a product. If the metal was detected in any of the twoor three samples of a product, then test results for that
product that were below the MDL were assumed to havea concentration of half the MDL. If the metal was notdetected in any of the samples tested of a product, weassumed a concentration of zero for all the samples ofthat product.

Non-Cancer Health Risks From Estimated Daily Intakes of Inorganic Arsenic, Cadmium, Lead, and Methyl Mercury
S E L EC T E D H E A LT H - B A S E D E X P O S U R E A N D OT H E R L I M I T S

Heavy Metal

SourceEPA (1991)
Inorganic Arsenic (currentlyunder review)

Endpoint and Basisfor Limit

Value (Unit)

Non-cancer oral reference dose based on
hyperpigmentation, keratosis, and possible vascularcomplications from chronic human exposures

0.3 ug/kg-day(upper limit)

Lower bound of suggested range whereby
0.8 ug/kg/day was the upper bound

0.1 ug/kg-day(lower limit)

Cadmium

EFSA (2009)

Based on a tolerable intake of 2.5 ug/kg bw/week,
to keep the population U-Cd concentration below
1 ug/g creatinine by the age of 50 years

0.36 ug/kg bw/day

Lead

OEHHA (2017)

OEHHA Proposition 65 Maximum Allowable DoseLevel (MADL) for Chemicals Causing Reproductive
Toxicity (total lead, oral exposure)

0.5 ug/day

FDA InterimReference
Levels (IRLs)

In 2018 FDA derived the IRLs from the CDC
reference value of 5 ug/dL of BLL to result in 3 ug/
day for children and 12.5 ug/day for women ofchildbearing age or WOCBA, to serve as usefulbenchmarks in evaluating the potential for adverse
effects in dietary lead.

Methyl Mercury

EPA (Updated2001, currently
under review)Non-cancer oral reference dose based on
developmental neuropsychological impairment

Salmonella

FDA

We compared our estimated daily intakes to healthbased limits in the above table using the following
equation: Hazard Quotient (HQ) = Estimated Daily Intake(or Exposure Dose)/Reference Dose.
If the HQ is greater than 1, then adverse health effects
are possible. As evidence suggests that impacts on

3 ug/day(children)
12.5 ug/day(WOCBA)
0.1 ug/kg-day
Absence in 25g

neurodevelopment are a sensitive endpoint for each of
inorganic arsenic, cadmium, and lead, we summed theHQs for the three heavy metals, to provide an estimateof the total hazard associated with a specific exposure
pathway, which is known as a hazard Index (HI). As with
HQ, if HI is greater than 1, then adverse health effects
are possible.

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