Hindawi Publishing Corporation

International Journal of Analytical Chemistry

Volume 2015, Article ID 164974, 7 pages
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Review Article
Recent Developments in Sweat Analysis and Its Applications

Saima Jadoon,1 Sabiha Karim,2 Muhammad Rouf Akram,3 Abida Kalsoom Khan,4
Muhammad Abid Zia,5 Abdul Rauf Siddiqi,6 and Ghulam Murtaza7
1
Department of Natural Resources Engineering and Management, University of Kurdistan, Hewler 44003, Iraq
2
University College of Pharmacy, University of the Punjab, Lahore 54000, Pakistan
3
Department of Pharmacy, University of Sargodha, Sargodha 40100, Pakistan
4
Department of Chemistry, COMSATS Institute of Information Technology, Abbottabad 22060, Pakistan
5
Department of Chemistry, University of Education, Attock Campus, Attock 43600, Pakistan
6
Department of Biosciences, COMSATS Institute of Information Technology, Islamabad 45320, Pakistan
7
Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad 22060, Pakistan

Correspondence should be addressed to Ghulam Murtaza; gmdogar356@[Link]

Received 13 November 2014; Revised 4 February 2015; Accepted 5 February 2015

Academic Editor: Frantisek Foret

Copyright © 2015 Saima Jadoon et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Currently, the clinical use of sweat as biofluid is limited. The collection of sweat and its analysis for determining ethanol, drugs, ions,
and metals have been encompassed in this review article to assess the merits of sweat compared to other biofluids, for example, blood
or urine. Moreover, sweat comprises various biomarkers of different diseases including cystic fibrosis and diabetes. Additionally,
the normalization of sampled volume of sweat is also necessary for getting efficient and useful results.

1. Introduction free fraction of drug from plasma to sweat through lipid

bilayer [3–5]. The main content of sweat is water (∼99%)
Similar to the sebaceous glands or hair follicles, sweat glands [2]. Besides, small amounts of the following substances are
are epidermal appendages that are normally distributed over also present: nitrogenous compounds such as amino acids
the whole body, excluding the nipples, lips, and external and urea [6]; metal and nonmetal ions such as potassium,
genital organs. Sweat glands are involved in perspiration sodium, and chloride ions [3]; metabolites including lactate
and act as excretory organ like kidney and lungs for drugs and pyruvate; and xenobiotics such as drug molecules [2].
and their metabolites, as shown in Figure 1. On the basis of In disease state, sweat may contain different ingredients as
morphology or secretory mechanism, there are two types biomarkers of the particular disease [2, 3, 6]. Blood, urine,
of sweat glands, that is, eccrine and apocrine glands. Sweat saliva, and sweat are the biofluids, which are used for clinical
glands allow the secretion of sweat through or without analyses such as pharmacokinetics study. Due to the presence
pinching off of outer cell parts [1]. Sweat is normally a of very nominal impurities, the sample preparation of sweat is
transparent biofluid with low tonicity and slightly acidic very easy as compared with other biofluids. In addition, sweat
nature with mean pH 6.3, that is, more acidic than blood [2]. samples are less prone to adulterations; thus such samples
Thus, basic drugs preferably accumulate in sweat than blood, can be stored for long periods [7]. Unlike other biofluids,
based on pH partition theory [3]. The transport of water sweat possesses excellent features including its noninvasive
insoluble drugs between blood and other biofluids depends sampling. Owing to this characteristic, sweat analysis is
on the pH of the other biofluids and the drug’s pKa which are considered as a rapid and easy process in comparison to other
helpful in theoretical computation of the biofluid-to-plasma biofluids, especially blood. Blood sampling is an invasive
concentration ratio of drug using Henderson-Hasselbalch procedure; that is, it needs surgery. Patients, who require
equation [3, 4]. The concentration gradient between plasma frequent analysis, are therefore at higher risk of infection.
and sweat provides driving force for passive diffusion of the In addition, blood samples are further processed for plasma
2 International Journal of Analytical Chemistry

Xenobiotics Together with a low intensity electrical current (∼3.0 mA)

and drugs applied for approximately five minutes, pilocarpine is applied
on small area of leg or arm for induction of sweating [37].
Phase I metabolism

Intermediate 2.2. Sampling of Sweat. An ideal sampler is the one that

metabolites is user-friendly and harmless to skin and quickly collects
Phase II metabolism
the sweat in sufficient volumes. Various versions of sweat
samplers have been designed and used for sweat sampling.
Excretory The simplest sampling technique involved the use of an
metabolites Bile Faeces occlusive patch consisting of 2-3 layers of filter paper or
gauze. However, this approach was time-consuming and
difficult to adopt owing to large size of patch. Moreover, these
patches were nonadjustable to skin [38]. The variation in
Serum/plasma/blood
sample pH and irritation to skin were other drawbacks of this
method [18]. To avoid skin irritation, nonocclusive device was
prepared using Whatman filter paper patch fixed on surgical
Urine Kidneys Saliva
dressing film lined with an adhesive layer for adjusting to arm
skin. This patch was safer to the underneath skin because
Lungs Skin of selective transfer of water, oxygen, and carbon dioxide
through this semipermeable film, which hindered pene-
tration of nonvolatile substances [38]. Since these patches
Breath Sweat
allowed evaporation of water from concentrated sweat leav-
ing sweat components such as chlorides, the entire volume of
Figure 1: Routes of excretion of various products after liver metabo- secreted sweat was not known. Resultantly, from the collected
lism.
samples, the results for cystic fibrosis (CF) test were not
authentic. Later on, the problem of water evaporation was
solved by using air- and water-tight, sweat collection bag
protein removal. In addition, the preparation of sweat sample connected with adhesive rubber. This advanced device was
is easier than that of urine. This merit of sweat as biofluid capable of retaining the whole volume of secreted sweat [39].
is particularly important in doping control where results are Moreover, the design of this device was further modified by
urgently needed [8, 9]. Beside these advantages, clinical use of connecting it with glass rollers, pipettes, and holders to make
sweat samples is presently limited due to high costs [10], time- it suitable for collection of sweat secreted in limited volumes
consuming sampling, infectivity hazards, and the need of [40].
volume normalization [11]. In addition, the rigorous attention For efficient analysis, the sweat samplers are usually asso-
should be paid to the analysis of metabolic products present ciated with the analytical instruments [41]. Macroduct and
in sweat [12]. Megaduct are the most frequently used commercial samplers,
Due to increased interest in the clinical use of sweat, available in different volumes (15 𝜇L–0.5 mL). Macroduct and
various approaches for sweat sampling and analysis have been Megaduct are used for collection of smaller (15 𝜇L) and larger
documented. Thus, the objective for preparing this review (0.5 mL) volumes of sweat, respectively. Both tools contain
article involved the summarization of various apparatus and a concave disk and a spiral plastic tube for sweat collection
techniques used for sampling of sweat and its analysis. [42]. Another modern sweat-sampling device is microstrip
Furthermore, the applications of sweat in clinical settings impregnated with a dye pH indicator that is a smart-phone
have also been discussed. application for in situ colorimetric testing [43].

2. Sweat as Biofluid 2.3. Sample Preparation for Analysis. Generally, sweat is

directly analyzed; however, sweat samples can further be
2.1. Induction of Perspiration. Apart from sampling and anal- processed if lipid or protein moieties are detected in sweat.
ysis, the induction of perspiration is a distinct phenomenon After sample collection, various procedures, including liquid-
unlike other biofluids, which are rather directly collected. The liquid extraction and solid-phase extraction, for the prepa-
physiological factors which enhance perspiration for getting ration of samples are used. Liquid-liquid extraction involves
certain volume of sweat include exercise and stress, while the use of analyte-specific solvents (e.g., aqueous phosphate
reduced perspiration is observed in cold [1]. For receiving buffer and organic solvents, e.g., methanol and acetonitrile)
sweat volume adequate for subsequent analysis, there are [37]. Solid-phase extraction involves the use of cartridges that
many factors which induce perspiration for sampling objec- internally consists of copolymers [8]. Sometimes the drugs,
tive; these factors include environmental factors (such as xenobiotics, and electrolytes are adequately discriminatory
temperature and relative humidity), body regulatory systems at the detector, and then their derivatization is also carried
(hormonal and sympathetic nervous system), diet, and cer- out during sample preparation for appropriate detection or
tain sweat inducing chemical compound such as pilocarpine. separation [44].
International Journal of Analytical Chemistry 3

2.4. Analysis of Sweat. The quality of sweat analysis depends resulting in formation of sticky mucus in various organs such
on the efficiency of sample collection and the accuracy and as lung, intestine, and other organs. This condition leads to
sensitivity of analytical methods [2–5, 9, 42]. Currently, serious repeated infections of the pancreas affected organs.
different analytical approaches have been used for the analysis Moreover, male infertility and dehydration are also observed
of unmetabolized drugs in sweat; however further focus of in CF patients. The sweat analysis for sodium/chloride ratio
researchers is needed for studying drug metabolites present (sodium and chloride contents) can, therefore, be useful in
in sweat normalized with its volume. Most of the sweat ana- diagnosing CF. In particular, chloride level is disturbed in
lyzers work on the principle of potentiometry, colorimetery, CF due to mutation in CFTR, and thus sweat chloride can
conductivity, or osmolarity [43]. be referred to as the biomarker for CF diagnosis [51, 52].
On coupling with mass spectrometers (MS), capillary On the basis of this biomarker, there are two types of CF,
electrophoresis and chromatographic approaches such as that is, typical and atypical CF. With a minimum of one
liquid-chromatography (LC) and gas-chromatography (GC) phenotype appearance, chloride contents ≥ 60 mmol per liter
are valuable for high-resolution separation of drugs or com- of sweat indicate typical (real positive) CF while atypical CF
plex metabolites in sweat. For drug analysis in sweat, the is manifested with chloride concentration in borderline range
most frequently used equipment is GC-MS coupled with of 30–60 mmol per liter of sweat. The normal (and border-
electron impact ionization [7–9, 12, 37, 44–46]. Moreover, line) concentrations of chloride in sweat are <30 mmol/L (30–
LC-MS/MS coupled with electrospray ionization has also 59 mmol/L) and <40 mmol/L (40–59 mmol/L) in infants and
been successfully used [37, 47] to determine drug concen- elders [51]. Moreover, sweat potassium as another biomarker
tration in sweat. In this context, immunoassay approaches is the focus of current research for early diagnosis of CF
including radioimmune analysis and ELISA are also being that assists in treatment efforts for the diseased person;
employed [8, 12, 37, 45]. Table 1 describes some approaches however clinical application of this new biomarker is still
for separation and detection-determination of drugs of abuse under investigation [53].
in sweat [8, 10, 13–19].
On the basis of sweat analysis, diabetic biomarkers have
To analyze sweat, the analytical instrument is selected
also been reported such as the average change in sweat
in accordance with the nature of target analyte(s) such
rates [54], composition of human sweat [40], and correlation
as sodium or chloride ions [48]. For single moiety analy-
between sweat glucose and blood glucose. The later approach
sis, some of the most frequently used dedicated analyzers
are the potentiometric Orion skin ion selective electrode produces promising results provided sweat gains no glucose
(ISE) for chloride (Orion Research, Cambridge, MA), or from environment [55]. This diagnostic technique involves
the colorimetric Scandipharm CF Indicator System chloride the analysis of foot sweat using a simple, reproducible
patch (Scandipharm, Birmingham, AL), the Wescor Sweat- indicator test [56] on the basis of color change of a patch from
Chek conductivity analyzer (Wescor, Logan, UT), and sweat blue (due to the presence of anhydrous cobalt-II-chloride) to
osmolarity analyzer (Nikon Research, Cambridge, ML) [49]. pink at 10 min on adding 6 water molecules.
Due to variable volumes of sweat samples, the normaliza- Jurado Gámez et al. have proposed sweat-based diag-
tion of sweat volume is also necessary for getting authentic nostic assay for lung cancer by discriminating between
results [6]. For normalizing the sweat volume, Appenzeller metabolomics of healthy and diseased subjects. In this
and coauthors introduced the concept of internal standard promising approach, sweat is diluted with 0.1% formic acid
by using and determining the level of sodium and potassium followed by the injection of sample into LC-TOF/MS which
necessitates only 10 𝜇L of sweat [57].
using capillary zone electrophoresis linked with diode array
Genomics and proteomics have played a key role for
detector set at 214 nm [39]. However, a study has suggested
searching sweat biomarkers such as dermcidin (DCD). Sweat
that sodium ion concentration is more suitable for using
contains DCD, a peptide containing 47-amino acids, which
in the normalization of sampled volume of sweat than
possesses antimicrobial activity against different pathogens
potassium ion concentration [50]. However, the concept of
in high salt concentrations and over an extensive pH range
normalization of sampled volume of sweat has not yet been
resembling to the human sweat. For this reason, sweat is
applied for the diagnosis of CF.
considered to be crucial for human skin microflora [58].
Moreover, DCD and the receptors for DCD are present
3. Applications of Sweat Analysis and overexpressed on the cell surface of invasive breast
carcinomas and their lymph node metastases and neurons
3.1. Diagnosis of Diseases. Since last three decades, much of the brain. These findings reveal that DCD is involved
attention has been paid towards application of sweat in in tumorigenesis by promoting cell growth and survival in
disease diagnosis. The best example of a disease diagnosed breast carcinomas [59]. Another prognostic biomarker is
through sweat analysis is cystic fibrosis (CF). This disease prolactin inducible protein (PIP) which is expressed in many
originates from genetic transformations in CFTR proteins exocrine tissues including sweat glands and is overexpressed
(cystic fibrosis transmembrane conductance regulating pro- in metastatic breast and prostate cancer [60]. In addition,
teins) in the sweat gland. The CFTR proteins are, nor- prognostic biomarkers have also been investigated in a study
mally, responsible for the transport of sodium and chlo- performed on eccrine sweat in healthy and schizophrenic
ride (transportNa-Cl ) in epithelial secreting cells. The genetic patients. The eccrine sweat contains plenty of various proteins
modification of CFTR causes the change in transportNa-Cl and peptides unlikely to that of serum showing that eccrine
4 International Journal of Analytical Chemistry

Table 1: Some approaches for separation and detection-determination of drugs of abuse in sweat.

Limit of
Number Analytical approach Examples of some analyzed drugs quantification References
(ng per patch)
Cocaine, codeine, 6-acetylcodeine, morphine,
1 GC-MS (electron ionization) 5–10 [8]
6-acetylmorphine, and heroin
Cocaine, codeine, 6-acetylcodeine, morphine,
GC-MS (electron ionization) 5 [13]
2 6-acetylmorphine, and heroin
GC-MS (electron ionization) Methadone 50 [14]
Cocaine, codeine, 6-acetylcodeine, morphine, and
3 GC-MS (electron ionization) 50 [15]
heroin
4 GC-MS (electron ionization) Codeine, morphine, and 6-acetylmorphine 2.5 [16]
5 GC-MS (electron ionization) Cocaine and heroin Not mentioned [17]
6 GC-MS (electron ionization) Cocaine, codeine, morphine, and 6-acetylmorphine 2.5 [18]
ELISA and GC-MS (electron
7 Codeine, morphine, 6-acetylmorphine, and heroin 3–5 [10]
ionization)
LC-MS-MS (electrospray
8 Fentanyl 0.09 [19]
ionization)

sweat may produce distinctive disease-linked biomolecules sweat concentrations of some metals (e.g., cadmium and
[6]. lead) or their cations, salts, or complexes are sometimes
comparable to those of urine; thus sweat can be used as a
3.2. Assessment of Drugs and Ethanol in Sweat. Currently, biofluid alternate of urine, particularly in some kidney disease
sweat analysis for drug contents is accomplished through two [29, 34]. Table 2 elaborates the studies of metal excretion in
approaches, that is, early and late testing. First methodology sweat conducted in different countries. It can therefore be
involves the immunochromatographic testing for qualitative stated that perspiration is a potential route for the excretion
detection of recently used drugs (within 24 h) involving of toxic metals from the body.
sweat sample collected at single time point for identifying
the individuals who are under the effect of drugs. Second 3.4. Assessment of Volatile Organic Compounds in Sweat.
methodology involves the patch technology for qualitative Large number of different compounds has been recognized
detection of previously used drugs (within 168 h) involving in human sweat, out of which >500 compounds are volatile
sweat sample collected at single time point for the follow- in nature [67]. Because of heterogeneous distribution of
up of drug users under treatment to substantiate abstinence various sweat glands in skin, the profiles of volatile organic
[3, 61, 62]. compounds (VOC) are different in different body regions,
Together with urine, sweat is an ideal sample for doping which also affect the odor of an individual [68, 69]. Moreover,
control. The volume of sweat perspired by the whole human VOC from personal care products and sweat may also
body in one day is 300–700 mL. This biofluid contains a interfere with each other during sweat analysis [70, 71]. In
small but quantifiable percent of a drug [63] excreted through addition, compounds which are volatile at body temperature
transcellular and paracellular pathways in skin [11, 64]. The are directly collected, while the other substances are obtained
reported drugs excreted through sweat in a quantifiable frac- through volatilization of collected sweat.
tion are the opiates, buprenorphine, amphetamines, gamma
hydroxybutyrates, cocaine, and cannabinoids [9, 65]. In
addition, ethanol contents in sweat as a function of time have 4. Conclusion
also been successfully analyzed after ingesting ethanol [51]. Based on sweat analysis, advancements in the genomics
and proteomics have enormously contributed to the field of
3.3. Assessment of Metals, Ions, and Salts in Sweat. Xen- metabolomics and the systems biology. The metabolisms of
ometabolomics is a branch of science that deals with the study the macromolecules in sweat glands produce lower molecular
of essential metals and xenometals in the organism contami- weight metabolites, such as the conversion of proteins to
nated through either ingestion of food or absorption through peptides or amino acids. Since, metabolomics deals with
skin by occupational exposures [66]. After getting into body, measurements of both precursor and metabolites, sweat can
some metals are converted to their xenometabolites (cations be used as a biofluid, in addition to blood and urine, to
or salts) followed by their solubilization in sweat. In addition explore biomarkers for various diseases. Subsequently, these
to excretion of metals as their free metals, ions, or simple discoveries help in exploring effective therapeutic moieties.
salts, excretion of some metals occurs in the form of their Since sweat consists of various biomarkers, these biomarkers
complexes. For example, lead complexed with high molecular have played an excellent role in diagnosis of cancer, diabetes,
weight compounds excretes through sweat [25]. The excreted schizophrenia, and cystic fibrosis. Conclusively, sweat can be
International Journal of Analytical Chemistry 5

Table 2: Studies of metal excretion in sweat.

Number Metals Country of study

References
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1 Canada Genuis et al., 2011 [20]
2 Mercury USA Robinson and Skelly, 1983 [21]
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20 USA Hohnadel et al., 1973 [33]
21 Tropics Omokhodion and Howard, 1991 [34]
22 Russia Parpaleı̆ et al., 1991 [35]
23 Germany Haber et al., 1985 [36]

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