Advancements and Challenges of Cigar Science, Testing and Regulation: A Review

Contributions to Tobacco & Nicotine Research
formerly: Beiträge zur Tabakforschung International
Volume 31 @ No. 2 @ July 2022
DOI: 10.2478/cttr-2022-0008
Advancements and Challenges of Cigar Science, Testing
and Regulation: A Review *
by
Richard Y. Abrokwah and Rana Tayyarah
ITG Brands LLC, A.W. Spears Research and Development Center, Greensboro, NC, USA
SUMMARY
On May 10, 2016, the U.S. Food and Drug Administration
(FDA) published a Final Rule that extended its regulatory
authority to all tobacco products, including e-cigarettes,
cigars, hookah and pipe tobacco (Deemed Products). Effec-
tive August 8, 2016, this decision greatly expanded the scope
of tobacco products being regulated by FDA and introduced
significant testing challenges that need to be addressed. The
major challenge for cigars in particular is testing as well as
generation of accurate and reliable data, in the absence of
certifiedreferenceproductsandstandardizedmethodologyfor
a product category with significant complexity and high
inherent variability. In this article, we provide an overview of
recent studies as well as active opportunities and on-going
challenges associated with regulating and testing cigars. To
the best of our knowledge, this is the first comprehensive
review of non-clinical research for this product category
(cigars). We are therefore convinced that, tobacco scientists
and farmers, analytical chemists, cigar consumers, tobacco
legal counsels, state and federal regulatory authorities will
find this review beneficial and insightful. [Contrib. Tob.
Nicotine Res. 31 (2022) 73–89]
KEYWORDS
Cigar tobacco, cigar science, cigar regulation, cigarettes,
HPHC, variability, machine-made cigar (MMC), premium
cigar
ZUSAMMENFASSUNG
Am 10. Mai 2016 veröffentlichte die US-Arzneimittel-
behörde FDA eine endgültige Regelung, mit der sie ihre
Regulierungsbefugnisse auf sämtliche Tabakprodukte
einschließlichE-Zigaretten, Zigarren sowie Wasserpfeifen-
und Pfeifentabak (einschlägige Produkte) ausweitete. Seit
ihrem Inkrafttreten am 8. August 2016 hat diese Ent-
scheidung den Umfang der Tabakerzeugnisse, die unter die
Regulierung der FDA fallen, stark erweitert und zur
Einführung beträchtlicher Prüfanforderungen geführt, die
es zu bewältigen gilt. Insbesondere bei Zigarren liegt die
größte Herausforderung in der Durchführung von Prüfun-
gen sowie der Ermittlung genauer und zuverlässiger Daten,
da es für diese Produktkategorie mit ihrer erheblichen
Komplexität und hohen inhärenten Variabilität an einer
standardisierten Methodologie sowie zertifizierten Refe-
renzprodukten fehlt.
In diesem Artikel geben wir einen Überblick über aktuelle
Studien sowie bestehende Handlungsmöglichkeiten und
Herausforderungen bei der Regulierung und Prüfung von
Zigarren. Nach unserer Kenntnis ist dies die erste umfas-
sende nicht-klinische Übersicht für diese Produktkategorie
(Zigarren).
Wir sind daher überzeugt, dass Tabakforscher und -erzeu-
ger, Analytiker, Zigarrenraucher, spezialisierte Rechts-
berater sowie regionale und nationale Regulierungs-
behörden diese Übersicht nützlich und aufschlussreich
finden werden. [Contrib. Tob. Nicotine Res. 31 (2022)
73–89]
*Received: 5th
November 2021 – accepted: 30th
March 2022
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License (CC BY-NC-SA 4.0).
© 2022 Authors who publish with this journal retain all copyrights and agree to the terms of the above-mentioned CC BY-NC-SA 4.0 license.

RESUME
Le 10 mai 2016, la Food and Drugs Administration publia
une règle finale élargissant son autorité de régulateur à tous
les produits du tabac, y compris les cigarettes électroni-
ques, les cigares, les tabacs pour pipe et pipe à eau (pro-
duits présumés apparentés). Prenant effet en date du 8 août
2016, cette décision se traduisit par un vaste élargissement
du champ des produits du tabac placé sous la régulation de
la FDA et souleva des défis manifestes en matière de test.
Le principal défi, dans le cas des cigares notamment, est la
conduite de tests ainsi que la production de données fiables
et précises, en l’absence de produits de référence certifiés
et d’une méthodologie normalisée pour une catégorie de
produits présentant une complexité significative et une
forte variabilité intrinsèque.
Le présent article livre un aperçu des études récentes ainsi
que des opportunités actives et des défis actuels associés à
la réglementation et aux épreuves applicables aux cigares.
A notre connaissance, il s’agit du premier tour d’horizon
complet et non clinique pour cette catégorie de produits
(cigares).
Nous avons, dès lors, la conviction, que les scientifiques,
les agriculteurs cultivant le tabac, les chimistes analystes,
les consommateurs de cigares, les conseillers juridiques de
la filière du tabac, les autorités fédérales et fédérées de
régulation jugeront cette publication utile et révélatrice.
[Contrib. Tob. Nicotine Res. 31 (2022) 73–89]
ABBREVIATIONS
ALCS Altria Client Services Center
CORESTA Cooperation Centre for Scientific Research
Relative to Tobacco
CPA Crop Protection Agents
CRM CORESTA Recommended Method
FDA Food and Drug Administration
GAP Good Agricultural Practices
GRAS Generally Recognized as Safe
GRLs Guidance Residue Levels
HCI Health Canada Intense
HPHCs Harmful and Potentially Harmful Constituents
ISO International Organization for Standardization
MLR Multiple-Linear-Regression
MMCs Machine made cigars
NNK 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone
NNN N-Nitrosonornicotine
PCR Principal-Components-Regression
PLS Partial-Least-Squares
RSD Relative Standard Deviation
SA Smoke Analysis
TTPA Tobacco and Tobacco Products Analysis
1. INTRODUCTION
A cigar is defined as a roll of tobacco wrapped in leaf
tobacco or in a substance that contains tobacco (other than
any roll of tobacco which meets the definition of ciga-
rette) (1). Despite the abundance of literature on the
composition of traditional conventional cigarettes, pub-
lished research is limited on the physical and chemical
properties of cigars. Interest in expanding fundamental
knowledge and standardization has increased in the last
few years. Hence, there has been a marked uptick in
activity from industry, academic and private laboratories
with regard to research and testing designed to better
understand cigar properties. For example, there were 27
presentations or publications tracked by CORESTA on the
topic in 2017, approximately the same number as the
previous ten years combined (2).
The overarching aim of this work has been to advance the
scientific knowledge of cigar tobacco content and the
resulting deliveries of select smoke constituents while
developing a foundational understanding of the inherent
variability of the product.
Cigars are combustible tobacco products consisting of
filler, binder, and wrapper derived from tobacco. Gener-
ally, cigars of all designs fall into two main categories:
Handmade/Premium cigars and machine-made cigars
(MMCs). Premium cigars are typically made from whole
tobacco leaves of a single tobacco type (dark air cured); are
hand rolled; are usually large, with burn times of up to
several hours; and are relatively expensive compared with
other tobacco products. For most premium cigars, unblem-
ished leaves are required for the wrapper. The binder is
also natural leaf and the filler is hand-rolled (i.e., not cut)
(3). Alternatively, MMCs are typically made using homog-
enized natural leaf wrapper, with or without binder, and
with cut tobacco for filler. MMCs are mass produced by
machines and may contain Generally Recognized as Safe
(GRAS) additives and/or non-tobacco components such as
a mouthpiece. In this paper we attempt to summarize and
comment on recent scientific efforts and analytical testing
standardization efforts by the industry, and to discuss
challenges and opportunities with regard to analytical
efforts for the product category.
2. METHODS
Approximately 1100 peer reviewed publications including
extant monographs were systematically compiled fromthis
subject-specific research. Digital data bases used to
identify and screen the articles were the CORESTA
website, University of California San Francisco library of
tobacco industry bibliographies, FDA website and google
scholar.On-lineresourceslikeComsol(www.comsol.com)
and Cigar Aficionado (www.cigaraficionado.com) were
also utilized. Public health related studies, cigar consump-
tion studies, unpublished manuscripts, thesis, dissertations
and newspaper articles were considered irrelevant for
meta-analysis and therefore excluded.
In situations where different articles from the same authors
were cited, the articles were scrutinized to ensure that the
data from each study was independent of each other and
without conflicts of interest.
Figure 1 shows a pictorial illustration of our method;
depicting the huge statistical differences that exist between
availability of published cigar literature, cigarette literature
and that of e-cigarettes. Our observations in Figure 1
highlight an extremely limited availability of cigar science
research publications, therefore literature as far back a
74 CTNR @ 31 (2) @ 2022

Figure 1. Statistical hierarchy of review method findings based on (a) general tobacco literature search and (b) cigar science
literature search for about 1100 peer reviewed publications.
1950 up to 2021 was utilized to capture inter-generational
scientific developments and milestones in the tobacco
industry. Key words/phrases searched were cigar science,
cigar tobacco, cigar regulation, cigar method development,
cigar chemical analysis, cigar tobacco variability, cigarette
tobacco, machine-made cigars, premium cigars, handmade
cigars, cigar smoke constituents and cigar tobacco farming.
All the articles were collated using the EndNote referenc-
ing tool (https://endnote.com/).
3. SOURCES AND ATTRIBUTES OF CIGAR
VARIABILITY
3.1 Demographic variability of cigar tobaccos
All testing, whether content or yield related, are impacted by
the tobacco and ultimately the growing conditions of that
tobacco. There have been recent efforts to increase under-
standing in this area for cigar tobaccos. Some have argued
that testing and reporting multiple constituents in cigar leaf
andsmokewithouthavingin-depthknowledgeofwhatdrives
the variability/variations will engender the submission of
somewhat valueless and inconsequential data to the regula-
tory institutions (4). Generally, controls to minimize year-to-
year variability from seed planting and harvesting to the
finished tobacco leaf remain a challenge. Variability in cigar
tobacco is a well-known issue and FDA has acknowledged
that blend changes due to “natural variability” do not require
a product to undergo premarket review 1
.
Cigar tobaccos, like cigarette tobaccos, have defined catego-
ries (such as dark air-cured and sun-cured tobaccos). Within
each of the categories are numerous sub-types such as
Sumatra, and Jatim), and varieties (such as Vuelta Abajo).
Unlike cigarette tobaccos, cigar tobaccos have little to no
standardization and are typically local varieties produced
from suppliers and even farm-based selections. Unlike
cigarette tobaccos, there are limited varieties produced
through anytypeofseedcertification process. So, though the
total number of cigar tobacco varieties is much lower than
that of cigarette tobaccos, standardization is significantly
lower for cigar seeds and the range in seed sub-types and
varieties is much greater (3, 4). In addition, the soil and
climate conditions of the growing area are significant
factors impacting variability of cigar tobacco physical and
chemical properties (5). Knowledge of the relationship
between the different types of soil, climate and the varieties
of crops allows tobacco breeders to produce and distribute
seeds specifically adapted to specific growing locations.
For example, LUNDH affirmed that the strength, elasticity,
thickness and shining quality of cigar wrappers strongly
depends on the type of soil and climate in which the
tobacco seed is planted. He claimed that even when
Nicaraguan seed is planted in Ecuador, the tobacco wrap-
per produced is very different from a native Nicaraguan
wrapper. He explained that the humidity from the constant
cloud cover in Ecuador yields firm and elastic wrappers
while the volcanic soil type in Nicaragua yields wrappers
that are less elastic (6).
LINDEGAARD reported results for a controlled study whereby
the same dark air-cured tobacco seed was planted in the same
crop year by two different farmers in the same country and
local area. A very significant natural variability was quanti-
fiedincludinga379%differencebetweenthearseniccontent,
115% difference in N-nitrosonornicotine (NNN) levels, 53%
difference in ammonia content, 31% difference in
4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)
levels,12%differenceincadmiumcontentand11%variation
in the nicotine content (4).
LINDEGAARD’s findings accentuated the existence of an
innate variability within the cigar product due to variations in
the tobacco itself. Additionally, MUKOYI et al. advised
researchers, investors, leaf merchants, policy makers, and
other stakeholders to trade and invest in the viable high-
quality cigar wrappers, binders, and filler grown in Zimba-
bwe. They reported that conditions in the Burma valley
area in Zimbabwe, such as ideal loamy to loam-clay well
drained soils, high geographical altitude, high ambient
relative humidity > 65%, high temperatures (28–35 °C),
and high rainfall > 1000 mm/week, promote the growth of
pliable fine textured, unique wrapper leaf, which favors the
design of premium cigars. They demonstrated that the soil
and environment in that precinct are also conducive to
performing agronomic germ plasm holistic research to
produce a various preeminent shade cloth-cured tobacco
1
Deeming Tobacco Products to be Subject to the Federal Food, Drug,
and Cosmetic Act, as Amended by the Family Smoking Prevention and
Tobacco Control Act; Restrictions on the Sale and Distribution of
Tobacco Products and Required Warning Statements for Tobacco
Products, 81 Fed. Reg. 28,974, 28,996 (published May 10, 2016) (the
“Final Deeming Rule”).
75
CTNR @ 31 (2) @ 2022

leaves capable of resisting foliar diseases like frog eyes,
deformations from the scorching sun, droughts and hail-
storms (7).
The effect of soil nitrogen content has been shown to
impact certain quality attributes of Kentucky dark fire-
cured tobacco. SIFOLA et al. showed that increasing the soil
nitrogen impacted the brightness and increased “body”
(thickness, density, and weight) of the leaf (8).
Another research group led by BORGES performed
experiments to establish the optimum nitrogen
concentration for percent yield and quality of cigar tobacco
based upon leaf chlorophyll content (9). Other researchers
have found that nitrogen fertilizer impacted alkali metals
like potassium but not alkali earth metals like calcium and
magnesium (10). Over a threshold level of nitrogen,
however, leaf quality declined rapidly (11) and potassium
fertilizers may positively impact yield (12).
MONZÓN HERRERA in collaboration with the University of
Hohenheim, Germany, utilized greenhouse (hydroponic
and pot farming) techniques to examine the effects of some
micronutrients and macronutrients on the development and
growth on four Cuban dark tobacco varieties used for
“Habanos” cigars. Their extensive study showed that soil
types deficient in micronutrients like boron, phosphorus,
zinc, magnesium, potassium nitrogen and manganese
resulted in various detrimental foliar diseases and defective
stemgrowth patterns in the tobacco plant. They ascertained
that the soil nitrogen content strongly impacted the
composition of metallic cations in the green tobacco leaf.
In conclusion the group advised that (i) early application of
nitrogen is needed for intensive vegetative growth of the
tobacco plant on landscapes that have compact soils which
rarely undergo lixiviation of nitrates, and (ii) for farmers to
obtain the best yield and quality harvest the nitrogen
content must be increased to about twice the current
recommended amount required for soil fertilization (13).
3.2 Agricultural practices on cigar tobacco variability
There has been significant research conducted in the area
of Crop Protection Agents (CPAs) specifications and
application in tobacco farms as well as Good Agricultural
Practices (GAP). VANN and FISHER conducted a study on
the effects of three CPAs (azoxystrobin, butralin, and
flumetralin) on flue-cured tobacco grown in six different
areas in North Carolina, USA. The goal of the researchers
was to evaluate residue levels of the CPAs in order to
administer their proper application rates as well as to
determine the minimum preharvest interval application of
specific CPAs. They gathered their data based on
individual year, individual location, individual CPA, and
individual stalk position (lower, middle, and upper) of the
tobacco plant. They concluded that although residue
concentration of azoxystrobin was quite high compared to
butralin and flumetralin, azoxystrobin played a critical role
in the control of target spot (Rhizoctonia solani), a foliar
disease which was responsible for a 7% yield loss in North
Carolina in the 2013 growing season (14).
In 2017 the CORESTA Agro-Chemical Advisory
Committee (ACAC) developed and documented various
trials performed by different companies across the globe to
standardize and mandate specific Cigar Guidance Residue
Levels (C-GRLs) for dark air-cured tobacco. Some of the
mandates for farmers included strictly controlling
fertilization of the soil, proper leaf variety selections,
systematic curing strategies, proper topping and suckering
as well as optimizing the fermentation process of the cigar
leaves. To further bolster the standardization initiative,
CORESTA also launched the Agrochemical Residue Field
Trials Task Force (RFT-TF) which focused on the
development of new agrochemical candidates for setting
GRLs in terms of leaf quality and integrity to draw clear
distinction between cigar leaf and cigarette tobacco leaves.
The task force compared the yields and CPA residues data
at different stalk positions between two crop protection
programs (the local one, normally based on CPA
application relative to when the pathogen is present and the
worst-case scenario based on weekly CPA applications), to
confirm which was the most effective for eradication of the
three main fungal and insect-related tobacco diseases (15).
3.3 Design characteristics
Differences in conventional cigarettes typically result from
variations in tobacco blends and relatively small variations
in cigarette construction and physical dimensions such as
length, diameter and pressure drop (16). In the case of
cigars, the physical parameters vary greatly within and
across product categories. In fact, there are several
categories of cigars; each consisting of tobaccos that are
unique and different from each other. The two major
categories which are premium and machine-made cigars
(MMCs) are discussed below.
3.3.1 Premium cigar design characteristics
“Premium”, handmade, hand-rolled, (or long-filler) cigars
consist of whole tobacco leaves that, when rolled, run the
length of the cigar. Long-filler cigars are of a higher quality
than short-filler or medium-filler cigars and tend to burn
for a longer time. Most “premium” cigars are made entirely
of long-filler tobacco, wrapped in a quality natural tobacco
binder and wrapper. Tobacco is sorted, bunched, rolled,
molded, and pressed by hand. Finally, the outer wrappers
are added. During quality control evaluations, cigars are
color matched for packaging (3).
Figure 2 shows a premium cigar design with sections
labelled using typical vocabulary while Figure 3 shows the
layers/parts of tobacco leaf used for making cigars (17, 18).
Typically, the premium cigar body is composed of the
wrapper (outer tobacco), binder, and the filler (inner
tobacco). Generally, the wrappers are harvested from
plants cultivated under shade (shade grown tobacco)
whereas the fillers and binders are cultivated under full
sunshine (sun grown tobacco). The binders are leaves
selected from the lower part of the tobacco stemand should
be wide, large, and undamaged as possible. During manu-
facturing, the binder is rolled around the filler leaves and
are together referred to as the bunch. The filler leaves are
composed of three proportions or varieties (from bottom to
top positions of the stem, respectively) namely, volado
(filler itself, which mainly contributes to combustibility),
seco (dry, mainly contributes to aroma), and ligero (light,
mainly contributes to strength). During the cigar hand
76 CTNR @ 31 (2) @ 2022

Figure 2. Construction of a typical premium cigar product (17).
Figure 3. Strata, morphology and parts of tobacco leaves used for cigar construction. Colored “stripes” in the diagram represent
concentric/rolled layers of tobacco (18).
rolling formulation process, the ligero leaves are sand-
wiched between the volado and seco leaves (7, 19). The
wrapper leaf which is finally wrapped around the bunch
must have excellent pliability and elasticity.
Figure 4 (a) shows the art of hand-made premium cigars
and (b) an ideal cigar wrapper (15). It is reported and
putative that the filler contributes about 85% of the total
cigar weight, the binder 10%, and the wrapper the remain-
ing 5% (20).
3.3.2 MMC design characteristics
MMCs cover much more of a range in design complexity
and variables compared to premium cigars. The two broad
sub-categories are large filter (short-filler) cigars and
Medium-Filler cigars. Large filter (or short-filler) cigars
are MMCs that consist of chopped up tobacco leaves,
which are then rolled into cigars and have a conventional
acetate filter applied.
The tobacco in this category of cigars often comes from
pieces of the leaf that have been discarded during the
process of rolling “premium” or long-filler cigars. Large
filter cigars tend to burn hotter and quicker than their long-
filler counterparts. By using short-filler tobacco and
machines to aid in the cigar rolling process, manufacturers
can substantiallyincrease the volume of production relative
to a hand-made long-filler cigar.
Medium-filler cigars are MMCs that consist of tobacco
leaves, which are chopped into pieces larger than short-
filler. Typically, tobacco used in the head and body may
differ for an MMC product. These cigars may differ greatly
in parameters such as diameter, length, and shape. These
variables impact the combustion products, generation of
water, variation within cigars, and air flow within the
different cigar products.
Due tothesebroaddesignvariablesforbothMMCs and hand-
made cigars, the standardized smoking regime was developed
to maintain a constant airflow through the cigar during
machine smoking rather than a constant puff volume as has
been specified with standardized cigarette smoking (21).
77
CTNR @ 31 (2) @ 2022

Figure 4. The art of hand-making premium cigars (a). The ideal cigar wrapper after curing in a barn (b) (15).
3.4 Cigars and cigarettes comparisons
Much of the recent analytical research for cigars has been
designed as a comparison to conventional cigarette results.
Someresearch has been focused on smoking perception but
much of the work has been in the area of comparing
analytical content and yield or product variability. Two
notable differences with regard to cigar smoke are (i) cigar
smoke tends to be more alkaline than cigarette smoke and
(ii) tobacco commonly used for cigars contains lower
levels of reducing sugars than the rapidly dried varieties of
tobaccos commonly used in cigarettes. Normal mouth
(buccal) saliva is known to be neutral or slightly basic. The
impact of the higher alkalinity of cigar smoke (pH 8.5) on
nicotine absorption has been studied by ARMITAGE and
TURNER. The group asserted that because of the higher
concentration of unprotonatednicotine in alkaline medium,
nicotine in cigar smoke is much more readily absorbed
through the buccal mucous membranes than the protonated
nicotine in acidic cigarette smoke (pH 5.3) (22). Extensive
research by LEFFINGWELL corroborated the complexityand
variability in the chemical composition of tobacco leaf
types (yellowed, flue-cured, Burley, Oriental, enzyme
treated, Virginia, etc.) grown in different parts of the globe
(23). They reported that Oriental leafexclusively contained
significant amounts of labdanoid (Z-abienol) and sugar
tetraesters, which impact sensory attributes of the Oriental
leaf. Quantitative examination of the cuticular tobacco
components of cigar leaf and other leaf by SEVERSON and
associates outlined striking differences in the levels of
analytes such as hydrocarbons, sucrose esters, docosanol
and diterpenes (24). Table 1 (adopted from the Tobacco
Monograph by HOFFMANN and HOFFMANN) points out
unique chemical composition differences between cigar
tobacco and four selected cigarette tobaccos. For example,
the table depicts that cigar tobacco contains less than 0.1%
polyphenols relative to tobacco blends used for conven-
tional cigarettes (1.6–5.1%) (25). Conversely the cigar
nitrate content appears higher than for cigarette tobaccos.
Additionally, the burn characteristics of the cigar products
differ fromconventional cigarettesbecausecigarstypically
burn “inside – out” (tunneling) as opposed to the “outside
– in” burn characteristics of conventional cigarettes (26).
Clearly one would expect that the pyrolysis product profile
for cigars will thus differ significantly from that of ciga-
rettes.
Table 2 identifies differences between selected volatile
components in the smoke of cigars, little cigars, and
cigarettes. The concentrations of nitrogen oxides (NOx) are
significantly higher in cigar smoke compared to cigarette.
This is attributed to the elevated nitrate content of the cigar
tobacco, the incomplete combustion, and the naturally low
porosity of cigar binders and wrappers (25). In contrast, the
ammonia content of cigar smoke is more than three times
less than the amount in cigarette smoke. The sources and
physical attributes (e.g., full length, filler length, weight,
etc.) of the cigars and cigarettes used in the above study are
defined in a previous investigation by HOFFMANN and
WYNDER (27).
3.5 Cigar tobacco chemical composition and variability
While testing of cigar smoke is more akin to in-use testing,
understanding of tobacco content and variability is an
important area of research with significant recent focus.
Several studies have focused on fundamental understand-
ing of the leaf. For example, LAUTERBACH and GRIMM
extensively investigated the chemistry of cigar wrappers
used on MMCs from various brands (28). They identified
biomolecules like deoxyfructosazines and fructosazines
(sugar-ammonia biomarkers) in the wrappers. They
proposed to further study the interactions of the biomarkers
and tobacco fillers. In a separate study, LIN et al. observed
that the chemical composition of cigar wrappers varied
significantly fromtopping to maturation of the tobacco leaf
wrappers. For example, during the first 21 days (from
topping to maturation) the ratio of total nitrogen to nicotine
decreased constantly whereas the sugar, calcium and
magnesium contents increased (29). These results are
consistent with composition from topping to maturation of
tobaccos used for cigarettes. The effect of curing tempera-
ture on the fatty acids profile and ability of amylase and
78 CTNR @ 31 (2) @ 2022

Table 1. Comparison of some selected components in the tobacco of cigars and four cigarette tobacco types (% of dry weight of
tobacco) adapted from HOFFMANN and HOFFMAN (25).
Component Cigar
Tobacco type used for cigarette
Burley Maryland Bright Oriental
Nitrate 1.4–2.1 1.4–1.7 0.9 <0.15 < 0.1
pH 6.9–7.8 5.2–7.5 5.3–7.0 4.4–5.7 4.9–5.3
Reducing sugars 0.9–2.7 1.5–3.0 1.2 7.0–25.0 5.5
Total polyphenols < 0.1 2.0 1.6 5.1 4.5
Nicotine 0.6–1.7 2.0–2.9 1.1–1.4 1.2–1.9 1.1
Paraffins 0.3–0.32 0.34–0.39 0.34–0.41 0.24-0.28 0.37
Neophytadiene 0.4–0.8 0.4 0.4 0.3 0.2
Phytosterols 0.14–0.16 0.3–0.39 0.38 0.3–0.45 0.26
Citric acid 5.5–6.0 8.22 2.98 0.78 1.03
Oxalic acid 3.3–3.6 3.04 2.79 0.81 3.16
Maleic acid 1.5–1.8 6.75 2.43 2.83 3.87
Table 2. Components in the gas phase of mainstream smoke of cigars and cigarettes, values are given for 1.0 g tobacco smoked
adapted from HOFFMANN and HOFFMANN (25).
Component Cigars Non-filter cigarettes Little cigars Filter cigarettes
Carbon monoxide (mg) 39.1–64.5 16.3 22.5–44.9 19.1
Carbon dioxide (mg) 121–144 61.9 47.9–97.9 67.8
Nitrogen oxides (NOx) (µg) 159, 300 160 45, 150 90–145
Ammonia (µg) 30.5 95.3 200, 322 98
Hydrogen cyanide (µg) 1,035 595 510, 780 448
Vinyl chloride (ng) n.a. 17.3, 23.5 19.7, 37.4 7.7–19.3
Isoprene (ng) 2750–3950 420, 460 210, 510 132–990
Benzene (µg) 92–246 45, 60 n.a. 8.4–97
Toluene (µg) n.a. 56, 73 n.a. 7.5–112
Pyridine (µg) 49–153 40.5 61.3 27.6, 37.0
2-Picoline, µg 7.9–44.6 15.4 17 14.8, 15.6
3- + 4-Picoline (µg) 17.9–100 36.1 32.9 12.6, 20.2
3-Vinylpyridine (µg) 7.0–42.5 29.1 21.2 102, 192
Acetaldehyde (µg) 1020 960 850, 1390 94.6
Acrolein (µg) 57 130 55, 60 87.6
N ’-Nitrosodimethylamine (ng) n.a. 16.3–96.1 555 7.4
N ’-Nitrosopyrrolidine (µg) n.a. 13.8–50.7 24.5 6.6
n.a.: data not available
invertase to regulate the carbohydrate content of cigar
wrapper leaf has also been reported (30, 31).
On the other hand, many of the recent tobacco studies have
focused on understanding content and variability of
analytes of regulatory concern. Typically, this is with an
underlying objective of determiningrelevance and feasibil-
ity of routine Harmful and Potentially Harmful Constituent
(HPHC) testing for this product category.
For example, LINDEGAARD studied dark air-cured cigar
tobacco of the same leaf grade (i.e., same seed, country,
local area, texture, color) grown by the same farmer from
2013 to 2015. He noticed significant variability in the
composition of ammonia, nicotine, NNN, NNK, arsenic,
and cadmium.
It is interesting to note that the difference in nicotine
concentration between 2013 and 2014 was 89% whereas
the difference in the same analyte between 2014 and 2015
was a nominal 2%. As discussed earlier, this researcher
found marked differences in tobacco analyte content in a
study wherein the same seed was planted in the same crop
year by different near-by farms (4). WAGNER et al. also
carried out a point-in-time variability study on the smoke
and tobacco. With regard to tobacco analysis, the focus of
the study was cigarette and smokeless tobacco HPHCs:
ammonia, arsenic, cadmium, NNK, NNN, and nicotine.
They inferred that for ammonia and nicotine, the %RSD
was the same for cigarettes and cigars at approximately
2%. However, for the other analytes, the %RSD of the
MMC fillers was twice that of the cigarettes (32).
TAYYARAH et al. tested different cigar products at multiple
laboratories for tobacco HPHCs. The choice of analytes
was based on FDA requirements for cigarettes and smoke-
less tobacco products since there were no specified HPHCs
for cigars at the time of the study (33). The design of that
study included evaluation of the results to compare range
of content and variability between cigars of different
79
CTNR @ 31 (2) @ 2022

design, by laboratories testing cigars fromthe same lots but
using their own methods. The group ensured that partici-
pating laboratories were ISO 17025 accredited and used
validated methods. With regard to tobacco analytes, there
was a clear difference in content for different cigars.
For example, nicotine ranged fromapproximately 8.3 mg/g
to approximately 30 mg/g. However, more interesting
findings from the study were that the reported values from
the different laboratories for the same samples were in
some cases different enough that, in a blind study, one may
conclude the results were from different samples. For
example, for Sample F, the tobacco NNN values reported
by Laboratory 1 and Laboratory 3 were in a similar range
at 1748 ng/g and 2050 ng/g, respectively. Laboratory 2
reported a value of 4497 ng/g for the same sample batch,
which was more than twice that of the other labs. The
differences in standard deviation of the nicotine values
between the laboratories were particularly conspicuous
with %RSD values of 0.8%, 14%, and 4% for Laboratory
1, 2, and 3, respectively. This supports the essence of
current initiatives to increase standardization of testing,
includingavalidated,internationally recognizedmethodol-
ogy. Other reported studies on tobacco constituents are
consistent with these findings. They include investigations
by KOSZOWSKI et al. wherein they described extensive
variability of the nicotine content and physical dimensions
of cigars and cigarillos in the cigar market (34). This
substantive difference in cigars has been studied and
confirmed by other research groups (35–38).
3.6 Physical parameters as a measure of inherent
product variability
Some researchers have focused attention on physical
parameters as a direct and practical measure of product
variability. Testing for weight and length is relatively
inexpensive with high throughput and low measurement
variability.
Testing for diameter and pressure drop may be less
reliable given the range of product designs and lack of
standardization for measurement technology; it is
advisable to limit comparisons of results for these
measures between products of different design and/or
between laboratories using different analytical methodol-
ogy.
WAGNER et al. found striking differences in magnitude
of weight and pressure drop (referred to as resistance to
draw in their work) when comparing a set of 10 hand-
made cigars, 77 machine-made cigars, and 10 typical
cigarettes. The researchers found that the relative weight
of 100 replicates varied as much as 48% for hand-made
products and 70% for machine-made products in the
study but was typically less than a 14% spread for
cigarettes (39). TEILLET,VERNON and COLARDpresented
findings from a study of diameter, length, weight, and
pressure drop for hand-made cigars. TEILLET reported
low variability for direct control measures (length and
diameter). However, measures not directly controlled
during the hand-making process had point-in-time
%RSDs of 40% (weight) and 120% (pressure drop) (40).
The findings highlighted in this review are supported by
additional studies conducted by other researchers (41–43).
3.7 Cigar smoke chemical composition and variability
There is a rich body of literature, inter-laboratory studies,
and significanthands-onexperience for testing constituents
of conventional cigarettes. For example, standard validated
ISO methods for analyses of polyaromatic hydrocarbons
(PAHs) tobacco-specific nitrosamines (TSNAs),
polyaromatic amines (PAAs), ammonia, chlorides, volatile
organic compounds (VOCs), “tar”, nicotine, and carbon
monoxide (TNCO) and metals in conventional cigarette
smokearewell-documented, establishedacrossthetobacco
industry, and in use in ISO-accredited third party labs. In
addition, several cigarette smoking regimes (ISO, HCI,
Massachusetts, CORESTA) and cigarette references have
been established, beginning as early as the 1960s (44). In
contrast, expertise and standardization with cigar HPHCs
testing is substantially limited. For instance, there is a
standardized puffing regime and handling requirements
(described in CORESTA Recommended Methods (CRM)
64 and 65) (21, 45), but application of that regime to cigars
for constituents beyond “tar”, nicotine, and carbon monox-
ide (TNCO) methods needs optimization for both method
development and testing consistency across labs. Within
the past decade, study designs, presentations, and publica-
tions have revealed the challenges encountered and strides
achieved in cigar testing method development. The chal-
lenges include optimization of smoke holder accessories
needed to accommodate different cigar sizes, lack of in-
house method development for cigar analysis and inter-lab
proficiency studies for both MMC and premium cigar
products.
Recent reports related to analytical testing of cigar
smoke have focused on understanding yield differences
across the product category, often in comparison to
conventional cigarettes, inherent variability of smoke
analytes, and challenges with regard to smoking parame-
ters and technology.
WAGNER et al. presented results for a TPM (total particu-
late matter) comparison between cigarettes and machine-
madecigars smoked under standard regimes. The cigarettes
were smoked using the standard regimes (ISO, HC (In-
tense)) and the cigars were smoked using the cigar smok-
ing CRMs referenced herein. The holder used for cigar
smoking, an ALCS smoke trap, was a custom design (39).
Figure 5 presents a striking example with the simplest
smoking measurement (the weight of trapped particulates),
to support observations of inherent product variability and
the challenges of machine-smoking cigar products (39). It
can be inferred from Figure 5 that the variability in TPM of
the cigars was substantially higher than the two cigarette
regimes. While the cigarette ISO and HCI smoking re-
gimes yielded approximately 5–25% and 30–70% TPM
variability respectively, the cigar variability was 40–120%
which was over 70% higher than that of the cigarettes. In
another study, TAYYARAH et al. compared mainstream
cigarette smoke analytes tested in different laboratories
using their own in-house methods. The analytes under
study were HPHCs typically performed on cigarettes such
as carbon monoxide, smoke nicotine, selected carbonyls,
VOCs,tobacconicotine,tobaccoammonia, TSNAs, PAAs,
and PAHs (33). Despite the fact that all the labs were ISO
17025 accredited and used properly validated methods,
80 CTNR @ 31 (2) @ 2022

Figure 5. TPM variability comparison for 146 commercial cigarette products and 86 commercial cigar products under different
smoking regimes , n = 55 (39).
their conclusions were similar to most findings for
tobacco analytes i.e., the analyte levels varied greatly
between samples and between reported results from
different laboratories testing the same product lots.
WAGNER et al. (32) also carried out a short-term vari-
ability study on the smoke and filler of 24 MMCs and
146 cigarettes products. They compared the variability
between MMCs and cigarettes of 19 selected HPHCs
under ISO, Intense and CORESTA smoking regimes.
Their results affirmed that, under a specific smoke
regime, variability between each quantified analyte was
about 5–20% more pronounced in cigars than in ciga-
rettes. For example, under ISO, the average %RSD of
NNK for cigars was approximately 22% relative to about
5% for cigarettes. Similarly, under ISO Intense, the
average %RSD of formaldehyde was 20% for cigars and
about 8% for cigarettes.
For cigars, the CORESTA regime produced the highest
HPHC variability while the ISO regime recorded the
highest HPHCs variability amongst cigarettes.
YOUNG et al. also investigated the extent of chemical
composition and weight variations within some selected
small (SM), large (LG), and sheet-wrapped (SW) cigar
products under the CRM 64 guidelines. Their results in
Table 3 summarize the changes in the mean %RSD that
occurred in the carbonyl yields from 2016–2017. The
values indicated represent the analytes that showed
statistically significant differences between 2016 and
2017. For example, in 2016 the mean %RSD of formalde-
hyde for Phillies Blunt (LG) was 9.6 and increased to
19.8 in 2017. This computes to a notable mean formalde-
hyde yield %RSD difference of about 106% within a
year. Even products of similar size that were tested in the
same year showed a wide range of %RSD values in
parenthesis. They inferred that since no certified/qualified
cigar reference was available to be used as a control, they
could not confidently attribute the relative %RSD values
to either the inter-sample or method variability (46).
3.8 Curing, pre-processing, aging and sampling on cigar
tobacco variability
The variability of tobacco-specific nitrosamines (TSNAs)
which occurs during flue-curing and air-curing of cigar
dark tobacco has been a contentious public health debate
and well-studied. Cultivation of dark air-cured requires
high quantity of fertilizers in nitrate NO3
!
form, which
produces high concentration of this polyatomic anion in
cured leaves. According to BUSH and coworkers, during
curing the aerobic conditions cause the reduction of the
nitrates to nitrites (NO2
!
) which then react with the
secondary alkaloids within the tobacco leaf to form the
TSNAs (47). RICHMOND et al. studied the correlation
between curing environment and TSNA accumulation in
two barns about 200 miles apart. They found that although
barn curing conditions like temperature and relative
humidity impacted concentration of TSNAs, other crucial
factors like the barn construction, inconsistency in
microenvironments within the same barn and improper
positioning of data loggers (which records/monitors the
temperature and humidity) could also introduce more
TSNA variability (48). Thus the impact of curing on
analyte variability is an important consideration.
Another source of variability within this product category
is sampling. The significance of monitoring analytes in
cigar tobacco via product sampling and sample size
considerations cannot be overemphasized. BORGES
MIRANDA et al. performed extensive characterizations of
both raw materials and cigar products to underscore the
inconsistencies of tobacco blends used for premium cigars
as well as the variability that arises from different testing
methodologies and analytes. In 2019, the group utilized the
near-infrared-reflectance-spectroscopic technique to
characterize 322 powdered samples (raw materials and
products) of dark air-cured Cuban cigar tobacco that were
processed the same way (19). Their study revealed signifi-
cant statistical differences between the total alkaloids-
81
CTNR @ 31 (2) @ 2022

Table 3. Carbonyl yields in cigarillo and leaf-wrapped cigar products tested in 2016 and 2017 under CRM 64 smoking regimen (n = 7)
adapted from YOUNG et al. (46).
Tobacco Product Brand Name
2016-Carbonyl yields, mean (RSD) 2017-Carbonyl yields, mean (RSD)
Tobacco
product
weight
(mg/unit)
Form-
aldehyde
(µg/unit)
Acet-
aldehyde
(µg/unit)
Acrolein
(µg/unit)
Tobacco
product
weight
(mg/unit)
Form-
aldehyde
(µg/unit)
Acet-
aldehyde
(µg/unit)
Acrolein
(µg/unit)
Cheyenne Cigarillo Dark
& Mellow (SM)
2462 (6) 11.6 (16) 1015 (8) 20.2 (22) 2688 (4) 8.9 (12)
a
1246 (16)
a
14.2 (54)
Cheyenne Cigarillo Dark
& Sweet (SM)
2354 (8) 10.2 (14) 1258 (12) 21.8 (21) 2806 (3) 9.8 (20) 1333 (13) 16.2 (25)
a
Dutch Masters Cigarillo (SM) 2484 (9) 16.7 (34) 2232 (9) 46.2 (30) 2879 (9) 9.8 (16)
a
2259 (23) 23.1 (32)
a
Game - Black (SM) 2161 (8) 16.3 (25) 1681 (10) 33 (22) 2363 (6) 12.1 (22)
a
1817 (14) 30.8 (30)
Swisher Sweet Cigarillos -
Sticky Sweet (SM)
2277 (5) 13.1 (11) 1551 (11) 33.6 (18) 2794 (2) 10.7 (17)
a
1571 (19) 22.5 (41)
a
Swisher Sweet Cigarillos (SM) 3048 (14) 16.1 (19) 1926 (10) 15 (43) 2682 (3) 12.9 (22) 1889 (15) 36.2 (31)
a
Swisher Sweet Cigarillos -
Black (SW)
2457 (3) 9.8 (24) 1548 (9) 25.7 (36) 2676 (3) 9.3 (18) 1799 (31) 20.7 (55)
Dutch Masters President (LG) 7538 (3) 11.8 (12) 4855 (7) 49 (16) 7603 (5) 16.3 (9)
a
3913 (17)
a
34.5 (22)
a
Phillies Blunt (LG) 6611 (6) 9.6 (15) 3152 (4) 35.8 (25) 6931 (4) 19.8 (18)
a
4145 (20)
a
64.6 (33)
a
Diameter at 15 mm: SM = 9–10.5 mm, SW # 8 mm, LG = 15–16.5 mm
a
indicates statistically different constituent yield for the tobacco product analyzed in 2016 and 2017 (p < 0.05)
nicotine, total nitrogen, and total ash concentration in the
cigar tobacco. They processed the spectra and evaluated
the variability of these analytes with several statistical
regression models such as the PLS, PCR, and MLR models
and ascertained that the PLS model exhibited better
reproducibility, precision, and prediction statistics. To
enhance standardization and mitigate some of the variabil-
ity in premium cigar tobacco, the group recently analyzed
about 3780 different cigars and proposed a chemosensory
technique and methodology for selecting raw materials
from specific lots and optimizing the aging time required
for processed tobacco leaves. Their study also identified
specific chemical constituents and independentvariables in
the raw materials that could be analyzed and used as
indexes of the cigar strength (49). BORGES MIRANDA et al.
have once again reported that it is consequential to sample
raw materials at the end of the stripping workshop, which is
the phase where the low-quality leaves are separated fromthe
production line for premium cigars. The group utilized a
randomized sampling design and three estimation errors
(difference between sample mean and actual population
mean) to measure the nicotine content of different batches of
tobacco produced in different geographical precincts. They
concluded that although about 2016 samples a year could be
analyzed (with an estimation error of 0.2 % w/w), the sample
count should be increased to include and account for leaves
pre-processed daily during the pre-processing season (50).
ODELIN and BORGES MIRANDA also inferred that the weight
of cigars had a substantial influence on the concentration of
the smoke analytes; and determined the minimal sample size
required to estimate the weight of a single premium cigar
(51). It is worth noting that CORESTA has embarked on
crucial studies as part of international standardization efforts
to address the TSNA variability and sampling predicament
(52, 53). Other researchers have focused on agronomic and
germplasm studies as means to standardize, optimize and
homogenize the cigar leaf composition at the end of pre-
processing to help minimize variability. In this regard,
MORÁN GÓMEZ et al. investigated the correlation between
bacteria genera population density, the pH and nicotine
concentration in cured tobacco leaves harvested from differ-
ent locations of the stalk/stem. They identified and isolated
bacterial microbiota such as Staphylococcus, Arthrobacter
genus and the Bacillus genus. They found that although the
Staphylococcus and Arthrobacter species are important
indicators, the Bacillus genera were the most predominant in
the leaves processed from all stalk positions. They also
inferred that the bacteria population density was more
dependent on the leaf nicotine levels than the changes in pH
values. The group further emphasized that genomic technol-
ogy could reduce the processing time of tobacco leaves,
improve the quality of lower grade leaves, and ultimately
promote a more homogeneous composition of the leaves at
the end of pre-processing (54). YE et al. conducted a similar
study using genetic sequencing to identify beneficial micro-
bial strains which could improve the quality of the cigar
products (specifically the aging process of the cigar product
itself) and hence reduce the end-product variability. Their
study revealed quite a significant diversity of fungi and
bacteria strains in ten different cigar products. The predomi-
nant bacterial genera were Staphylococcus, Acinetobacter,
and Pseudomonas while that of the fungal genera was
Aspergillus (55).
4. ANALYTICAL METHODS DEVELOPMENT AND
STANDARDIZATION EFFORTS
Several researchers have reported findings that shed light
on the challenges of analytical cigar smoking. These
include conditioning protocols for cigar products, the
effects of lighting technique on smoke constituents,
number of relights, effects of ash removal, and the
complexity of choosing a proper cigar holder (56–58).
82 CTNR @ 31 (2) @ 2022

Table 4. Selected analytical methods previously applied to testing of cigar leaf and cigar smoke constituents.
Sample analyzed
Constituent and method of
determination
Method feasibility with existing
equipment
Detection limit
Tobacco (1.0 g) from cigarettes
was placed into a 20-mL head-
space vial. Internal standard
solution (2 µL of 1 µg/µL 2,6-
dichlorotoluene) and flavor spike
mixture (1 µL of 1 µg/µL each
benzaldehyde, tetra-methyl-
pyrazine, methanol, and anethole
in ethanol) were added. The
samples were sealed and
allowed to equilibrate for 2 h at
room temperature before
analysis (59)
Flavor additives to tobacco (e.g.,
menthol, anethole, benzal-
dehyde, and tetramethylpyrazine)
Headspace solid-phase micro-
extraction-gas chromatography-
mass spectroscopy (HS-SPME-
GCMS) for both qualitative and
quantitative analysis)
Feasible but could be very
tedious, time consuming &
unproductive
Benzaldehyde = 66 ng/g
methanol = 120 ng/g
anethole = 16 ng/g
tetramethylpyrazine = 163 ng/g
acetophenone = 41 ng/g
10.0 g tobacco sample was
added to 40 ml dichloromethane.
Then the mixture was shaken
overnight and steam distillated
for 3 h to obtain 800 mL aqueous
solution of volatile components
using a simple apparatus (60)
Lactones, benzaldehyde, 6-
methyl-2-heptanone, 2,4-
dimethyl-1-penten-3-one, etc.
Steam distillation (SD),
simultaneous distillation and
extraction (SDE) and headspace
co-distillation (HCD)-GC-MS
utilized for all volatiles
Feasible but could be very
tedious, time consuming &
unproductive
Total detected
315.72–445.48 µg/g
Evaluation of volatiles from flue-
cured tobacco varieties, smoke
organoleptic (61)
Lactones, benzaldehyde,6-
methyl-5-hepten-2-one, etc.
Steam distillation of 10 g
tobacco, capillary GC/GC-MS
Distillation system must be
available
200–600 µg/g
Smokeless tobacco products
including snuff, plug tobacco,
chewing tobacco, pellets, and
snus (62)
α- and β-angelica lactones
Headspace gas chromatography
mass spectrometry (HS-GC-MS)
Feasible. However, reference
standards for β-angelica lactone
unavailable or difficult to obtain
The limit of detection was 30
ng/g and limit of quantitation
65 ng/g with a variability of
9–44% (RSD)
Tobacco samples used for
analysis were Brazilian flue-
cured,Kentucky Burley, N.
rustica, and Greek and a sample
of commercially available roasted
peanuts (63)
Benzaldehyde, 6-methyl-5-
hepten-2-one, acetone, hexenal
Chromatography-mass selective
detection-flame ionization
detection (PT-GC-MSD-FID)
hyphenated technique with
purge-and-trap-gas
Feasible with little modification Semiquantitative and qualitative
analysis
Qualitative and quantitative
analysis was developed and
validated for volatile flavour
components in flue-cured
tobacco (64)
Flavour components in flue-cured
tobacco (e.g., pyridine, 6-methyl-
5-hepten-2-one, benzene
acetaldehyde, benzaldehyde,
furfural)
HS-SPME followed by GC × GC-
TOF-MS
Feasible but must have TOF-MS
on scope
5.7–147.6 ng/g
Determination of selective
phenolic compounds in cigarette
and MMC cigar smoke (65)
Phenolics (e.g. hydroquinone,
resorcinol, phenol, catechol, and
o-, m-, and p-cresol).
Ultra-high pressure liquid
chromatography (UHPLC) and
fluorescence detector (FLD) with
a sub-2 µm pentafluoro-
phenylpropyl phase analytical
column
Feasible high throughput method
that is based on CRM 78, which
has a run time of 10 minutes
Quantitative and qualitative
analysis
Continued refinement and extension of standard analytical
methods and techniques along with establishment of
reference products is the primary response to these chal-
lenges (21, 45). Specific analytical methods and validation
protocols for cigars need to be developed. Listed in Table 4
are a summary of results for several studies which focused
on testing of constituents in cigar tobacco leaf and smoke
which could be adopted or further developed (59–65).
Several researchers have investigated the feasibility of
extension of cigarette smoking methods for use with cigars,
83
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Table 5. Cigar reference products available through the University of Kentucky (72).
Reference cigar Product type Cigar diameter (mm) Cigar length (mm)
1C1 Large machine-made cigar 15.9 136.5
1C2 Machine-made filtered cigar 7.8 99.0
1C3 Small machine-made cigarillo 11.0 109.5
1C4 Large machine-made natural wrapper 12.8 103.0
but this work has only confirmed the need for cigar-
specific smoking methods. For example, the CORESTA
Tobacco and Tobacco Products Analysis (TTPA) and
Smoke Analysis (SA) Sub-groups have formally taken this
as a primary approach to CRM development (66).
PREPELITSKAYA et al. investigated the feasibility of
analyzing the ammonia content of cigars using the already
standardized CRM 83 “Determination of Ammonia in
Mainstream Cigarette Smoke by Ion Chromatography” for
testing cigarettes. Their findings indicated that specialized
methods needed to be developed for the analysis of ammo-
nia in cigar smoke as the CRM 83 method had shortcom-
ings when applied to cigars (67). BROOKS presented a
method for volatile organics in cigar smoke using a
modification of an existing in-house method for cigarettes
(68). Separately, JABLONSKI et al. and BALLENTINE et al.
took a similar approach to developing a smoke carbonyls
analytical method for cigars (36, 69).
Studies are currently underway to evaluate cigar tobacco
leaves and smoke tested for HPHCs typically applied to
cigarette and/or smokeless tobacco testing like carbon
monoxide analysis, smoke nicotine, selected carbonyls,
VOCs,tobacconicotine,tobaccoammonia, TSNAs, PAAs,
and polyaromatic hydrocarbons. A typical example is
CORESTA Project 198 which is a collaborative study to
analyze BaPs and TSNAs in cigar smoke (70). For smoke
measurements,CORESTArecommendedandISOmethods
for conditioning, smoke collection, and TNCO analysis of
cigar tobacco products as described in CRM 64 and
CRM 65 have been employed. Details from these analyses,
along with information regarding challenges associated
with testing across a range of cigars have fairly been
investigated.
4.1 Cigar reference products
Another concern is that although there is availability of
multiple reference cigarettes, internationally approved
cigar testing/smoking references or monitors have not been
established.
Fortunately, a project led by an industry team and the
University of Kentucky in collaboration with CORESTA
and accredited tobacco testing facilities to develop cigar
monitors/references was completed in 2019 to fill this gap.
A set of reference products, described in Table 5, were
formulated with different tobacco composite blends and
with varying design features that can represent most of the
cigar shapes and sizes (71). The university has developed
and marketed several tobacco references, including RT6,
a flavored cigar ground filler, and RT8, an unflavored cigar
ground filler (72). The University of Kentucky was
recently awarded a U.S. federal grant to develop a set of
certified reference products (73). Nonetheless, until this
project is completed, gaps will exist in the literature for the
definitive comparison of physiochemical composition of
cigar tobacco leaf and smoke constituents. Additional
studies related to analysis of cigar smoke in the recent past
include work by DETHLOFF et al. and MUELLER and
COLARD, among others (74–78).
5. REGULATION
World-wide tobacco regulation is in various stages of
implementation along different strategic pathways.
Typically, cigars represent a small fraction of a country’s
tobacco market and have been a much lower priority for
regulatory actions than cigarettes. In most countries that
have implemented regulations, the focus has been on
physical measurements, ingredient and marketing reports.
In the USA, the FDA has taken an approach similar,
though delayed, to the approach taken for cigarettes. FDA
regulation of cigarettes and smokeless tobacco products
began in 2009 (79). Over time, an expanded list of
recommended HPHCs for those products has been
established. As previously noted, in 2016, the FDA
published a Final Deeming Rule extending its regulatory
scope under the Tobacco Control Act to all other tobacco
products, including cigars (80).
Once FDA publishes final guidance relating to HPHC
testing, the Final Deeming Rule as written will require
stand-alone HPHC testing data for cigars. While stand-
alone testing may be required for these products under the
Tobacco Control Act, the challenges discussed herein
related to the variability inherent to cigars make testing for
comparativepurposesunreliable. Accordingly, researchers
consistently urge caution against use of any such analytical
testing data as metrics for product comparisons in the
context of substantial equivalence review.
For instance, with regard to HPHC testing, LONG recently
enumeratedthechallengesassociatedwiththeproposedFDA
objective to use HPHC data as an analytical rubric to
determine the substantial equivalence (SE) for cigars. He
elaborated on an extensive study carried out by Tabacalera
USA (TUSA) using 91 premium cigars of 43 different sizes
and 18 different blends of dark air-cured tobacco, wherein
they inferred that almost all the 36,000 data points generated
were statistically misleading, inconclusive and disclosed the
immeasurable variability that existed even between cigars of
the same size as well as cigars made from the same tobacco
composite blends (81). In general, researchers emphasized
that, based on the relatively high inherent variability of
many analytes with unknown factors, it is advisable to
avoid cigar comparisons using HPHC testing (4, 81).
84 CTNR @ 31 (2) @ 2022

6. ON-GOING CHALLENGES AND ACTIVE
OPPORTUNITIES
First and foremost, researchers and regulators must under-
stand that there are certain challenges with this product
category that will always be a consideration for study
design, data analysis, and evaluation of data across the
product category. The inherent variability of cigar tobacco
due to uncontrollable agricultural considerations, along
with variability of the seed genome, and product construc-
tion cannot be mitigated with analytical controls or method
standardization. That said, there are many active and
potential opportunities in this area of testing.
For example:
• EstablishmentofISO standardizedanalyticalmethodolo-
gies for appropriate measures and analyses to properly
characterize cigars and cigar smoke across the spectra of
designs,
• Full characterization and consistent use of recent and
pending referencecigars and cigar tobaccos for surrogate
characterization studies, aging studies, and method or
laboratory comparisons,
• Increasing standardization with regard to smoking
equipment, physical parameter measurement require-
ments, cutting and measurement standards, lighting and
relighting techniques,
• Continued evaluation of the approach for collecting
mainstream smoke as applied to all cigar categories to
account for the significant differences in design parame-
ters. For example, design parameters like circumference,
length, mouthpiece-type, diameter determination of
cylindrical vs. non-cylindrical products, ventilation, and
raw components vary significantly across the portfolio of
cigar products,
• Improvements to conditioning and storage requirements
to allow greater consistency between laboratories, and
• Establishment of data reporting norms that allow for
consistent data analysis across the product category.
To address the challenges above, several approaches have
been undertaken, or are currently underway. In the absence
of standardized testing specific for cigars, several contract
testing laboratories have chosen to incorporate in-house
developed cigar methods into the scope of their ISO 17025
accredited methodologies for tobacco product testing. The
salient risk in these scenarios would be how to track cigar
testing as well as how to account for inter- and intra-
laboratory data/report reproducibility or uncertainties over
time. Within CORESTA, severalactiveworkinggroupsare
addressing these challenges for all cigar products. The
CORESTA active working groups acknowledge that there
are many different types of cigars and that one testing
methodology will not be appropriate for all cigar products.
For example, the CORESTA Cigar Smoking Methods Sub-
group is currently documenting and publishing the techni-
cal reports and technical guidelines associated with the
TNCO testing of a variety of cigar products (82). In
addition,threeCORESTARecommendedMethods(CRMs
46, 64, and 65) for conditioning and collection of smoke
from cigars have been revised to more accurately reflect
technology capabilities and applicability to a wider range
of cigar products (21, 45, 83). Further, a CORESTA
project to specifically address challenges for testing hand-
made long-filler cigars has recentlybeenconcluded. Lastly,
the CORESTA SA Sub-group and the TTPA Sub-group are
both actively seeking opportunities to include cigars in
inter-laboratory proficiencystudiesascapabilities to enable
standardized and uniform testing across all laboratories.
The TTPA Sub-group has brought cigars into scope for
nine tobacco methods with additional methods expansions
in progress (84). The SA Sub-group has recently completed
its first joint experiment for cigar smoke constituents and
has established a long-term plan for cigar CRM develop-
ment (85). The University of Kentucky has established
plans to expand the scope of their proficiency testing
program to include cigar testing (71, 73)
7. CONCLUSIONS
This review provides a summary of recent analytical
research in the area of cigar testing. Undeniably, relative to
cigarettes, there is much less research on cigars, hence
challenges envisaged with analytical testing of cigars
remain to be addressed thoroughly. Especially with regard
to the substantial variety between same cigars and cigars of
different brands as well as tobacco leaves of different
origin, year of harvest and/or method of cultivation.
However, there is consistency in the findings reported
herein, which underscores the fact that cigars have a very
high inherent variability which leads to a very wide range
of agricultural yields.
There has been significant on-going activity with regard to
cooperative methods of development and standardization.
Recent successes in this area have included establishment
of a set of reference cigars, establishment of guidance for
hand-made cigar testing, and strategies for expansion of
scope for standard or accepted methodology specific to
cigars. With regard to regulatory oversight, researchers
recommend against using HPHC testing for product
regulation,comparison,andcharacterizationduetothehigh
inter-andintra-productvariability.Physicalparametersand
ingredient reports seem most practical metrics for product
comparison given the high complexity and inherent vari-
ability of the product category along with the relatively
immature foundation of analytical standardization.
While the studies reviewed in this manuscript highlight an
increase in the volume of research associated with cigar
testing, additional standardization and cooperative testing
is needed to establish a true foundation of analytical
understanding of this product category.
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Corresponding author:
Richard Abrokwah
ITG Brands LLC
Greensboro
North Carolina, USA
E-mail: Richard.abrokwah@itgbrands.com
89
CTNR @ 31 (2) @ 2022

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