Green Analytical Chemistry

Sergio Armenta, Francesc A Esteve-Turrillas, Salvador Garrigues, and Miguel de la Guardia, Department of Analytical
Chemistry, University of Valencia, Burjassot, Spain
© 2020 Elsevier Inc. All rights reserved.

Past, Present, and Future of Green Analytical Chemistry 2

The Role of Analytical Chemistry in the Food Industry 3
Direct Analysis: Dreams Come True 4
Benefits of Greening Sample Preparation/Extraction 6
Green Metrics Tools 8
Future Trends and Efforts 9
References 9

List of abbreviations.
ATR Attenuated total reflexion
DESI Desorption electrospray ionization
DI-SPME Direct immersion solid-phase microextraction
DLLME Dispersive liquid–liquid microextraction
DSPE Dispersive solid-phase extraction
DSPME Dispersive solid phase microextraction
ED-XRF Energy dispersion X-ray fluorescence
EHS Environmental, health and safety
ETAAS Electro thermal atomic absorption
FAO-UN Food and Agriculture Organizations of the United Nations
FNME Fiber-packed-needle microextraction
Foodome Food domain
GAC Green Analytical Chemistry
GXLs Gas-expanded liquids HF-LPME Hollow fiber liquid-phase microextraction
HS Head-space
HSP Hansen solubility parameters
ICP MS Inductively coupled plasma mass spectrometry
ICP OES Inductively coupled plasma optical emission spectroscopy
LAj Laser ablation
LCA Life cycle assessment
LIBs Laser induced breakdown spectroscopy
MAE Microwave assisted extraction
MEPS Microextraction by packed sorbent
MIR Middle infrared
MS Mass spectrometry
NEMI National Environmental Methods Index
NIR Near infrared
OES Optical emission spectroscopy
PHWE Pressurized hot water extraction
PLE Pressurized liquid extraction
QuEChERS Quick, Easy, Cheap, Effective, Rugged and Safe
RDSE Rotating disk sorptive extraction
REACH Registration, Evaluation, Authorization and Restriction of Chemicals
SBSE Stir bar sorptive extraction
SCSE Stir cake sorptive extraction
SFE Supercritical fluid extraction
SPE Solid-phase extraction
SPME Solid-phase microextraction

Reference Module in Food Sciences https://doi.org/10.1016/B978-0-08-100596-5.22800-X 1

2 Green Analytical Chemistry

SRSE Stir rod sorptive extraction

SVHC Substance of very high concern
TBA Thiobarbituric acid
TVB N Total volatile basic nitrogen
TVC Total viable counts
UAE Ultrasound assisted extraction

Past, Present, and Future of Green Analytical Chemistry

The idea of clean and environmentally sustainable Analytical Chemistry was born in 1995 with the Editorial of de la Guardia and
Ruzicka (1995) entitled “Toward environmentally conscientious Analytical Chemistry through miniaturization, containment and
reagent replacement” published in Analyst. However, although the analytical community has been environmentally sensitive for
a long time, the impact of this first approach remained practically unknown for the scientific community till the development of
the idea of Green Chemistry, introduced by Anastas (1999), focused on chemical synthesis and chemical processes through catal-
ysis. The term of Green Analytical Chemistry was proposed by Namiesnik (1999), replacing older denominations of the same
concept such as clean, environmentally or sustainable methods. Up to now, Green Analytical Chemistry has been deeply presented
in a series of books (Koel and Kaljurand, 2010; de la Guardia and Armenta, 2011; de la Guardia and Garrigues, 2011, 2012). Since
then, a huge amount of scientific papers have been published upgrading conventional analytical methodologies following Green
Analytical Chemistry philosophy, including more than 100 review papers published in journals of different areas like chemistry,
environmental science, agricultural and biological sciences, and biochemistry, genetics and molecular biology (Scopus database,
2019). Moreover, A. Galuzska et al. enunciate in 2013 the 12 principles of Green Analytical Chemistry, using the mnemonic
SIGNIFICANCE, as guidelines and recommendation to green analytical methodologies encouraging operational issues; such as
promote direct analytical techniques, avoid sample treatment or derivatization step, reduce the number of samples to analyze,
promote in situ measurements, integrate analytical processes, work on automation and miniaturization, downsize required energy,
reduce reagent consumption and wastes, made multianalyte determinations in each sample, avoid the use of toxic reagents, and
facilitate the use of renewable reagents (Gałuszka et al., 2013). Present efforts have been focused on the development of novel
high performance analytical instrumentation and methodologies, but the reduction of environmental impact of chemical analysis
(such as energy consumption, use of toxic reagents, or generation of wastes) needs to be also considered. Thus, a compromise
between the analytical figures of merit and the associated risk for the environment and human life must be comprehensively studied
and provide the guidelines for future research on this field.
To understand the great success of Green Analytical Chemistry, it must be considered: i) the changes on the mentality of the
advanced society in the XXI century, ii) the development of national policies and international conferences regarding the protection
of the environment and iii) economic reasons.
Nowadays, there is an increasing concern of society about the deleterious side effects of human activities and, in this framework,
there is a need for a severe control of the production, use and fate of synthetic organics; the European Norm REACH (European
Parliament, 2006) is a good example since it provides the main ideas to the surveillance and control of chemicals.
On the other hand, it is clear that the increase of the world population, the extension of life expectancy and the problems created
by climate change have moved the conscience of our societies to improve the environmental care. This is an important reason for the
general acceptance of the green alternatives. Laws follow the sensibility of societies and, other than protecting the individual citizen
rights, it is perfectly reasonable the development of control measurements to avoid the effects of pollution on the human health.
This philosophy has been evolved from soft to hard position and not only thinking on the effects on the human health but also on
the future of the planet. So, to be adapted to increasingly strict rules, it is absolutely necessary to reinforce the control of the use of
chemicals and to avoid waste generation.
However, one of the main reasons for the success of Green Analytical Chemistry between industrial and laboratory managers is
the fact that, in most cases, the green alternative methodologies offer a reduction of direct costs through the reduction on energy and
chemicals consumption. Additionally, the minimization of indirect costs derived from the treatment of wastes also reduces total
analyst cost and it could explain the increasing interest on green analytical methods.
The ideal analytical method should be that able to perform direct on-site or remote measurements to solve the analytical
problem. However, this dream cannot be accomplished for most of analytical problems, and typically, a sample pre-treatment is
required for the determination of analytes or sample properties at trace concentration and/or in complex matrices, with the corre-
sponding drawbacks due to potential analyte losses or contaminations. These steps also increased risk to both, the operator and the
environment (Armenta et al., 2017).
The future perspectives in Green Analytical Chemistry are thus, focused to search for remote and direct control methodologies
based on vanguard screening methods (Valcarcel and Cardenas, 2005) and also for greening the rearguard methodologies that are
required in many cases for both, solving the problems or to obtain reference data to build screening models.
 Green Analytical Chemistry 3

In a control philosophy of food production and market, which can be summarized in the traceability expression “from farm to
fork”, nowadays it is also important to include details concerning the evolution of food components during stock and cooking prac-
tices and also their changes during the digestion process. So, aspects like proteomics, transcriptomics or metabolomics, all of them
included in the definition of Foodomics, are important aspects to be considered and to do so analytical methods are the main
concern and thus, once again, there is a need for greening these methodologies.
In short, green analytical methods should: i) reduce or avoid the deleterious side effects of classic ones, ii) improve the main
analytical features of the method and iii) reduce the time for making decisions, as the fundamental goals.
In previous sentences the focus has been put on the environmental aspects and, based on that, it is possible to introduce the so
called green parameters, as a part of the different aspects to be considered to evaluate alternative methodologies to make a determi-
nation. However, it must be also taken into consideration that the desired simplification of methods also involves a drastic reduc-
tion of error sources. To delete sample treatment could reduce the variability of results and risks of contamination and loses, thus,
offering advantages on precision and accuracy. However, the reduction of the analysis time is the main aspect to be considered to
improve decision making, and this aspect, regarding the food chain, is a major concern to avoid health risks.
Future developments of analytical methods related to foodomics should be oriented to the use of direct methodologies, and
when it cannot be achieved, a minimal sample treatment by using automatized, miniaturized, and low energy procedures, should
be employed, using non-toxic, renewable, and environmentally friendly reagents. Most of foodomics approaches involve the use of
liquid chromatography mass spectrometry techniques, thus the organic solvents consumption is one aspect that must be properly
evaluated. In this framework, the replacement of conventional systems by ultra-high performance liquid chromatography, which
use sub 2 mm particles diameter and operates at high pressures till 6000 psi, typically reduces one order of magnitude the solvent
consumption. Other strategies to reduce environmental impact of chromatography approaches are focused on the use of automated
switching using a mobile phase recycler that automatically detects and splits-up to waste the correspondent eluting peaks and sent
back clean mobile phase to the reservoir (Giaquinto and Samide, 2013), or the replacement of mobile phases by green solvents
obtained from renewable resources (Pena-Pereira and Tobiszewski, 2017) or high alcoholic strength drinks (Welch et al., 2010,
2015). As summary, Fig. 1 shows a scheme of a typical foodomics procedure and the green possibilities that can be achieved in
each step.

The Role of Analytical Chemistry in the Food Industry

Regularly, official food control systems are implemented to ensure food safety, to decide its adequacy for human consumption, and
to control that products are honestly and adequately labeled. Currently, food control systems vary from country to country

• In-situ analysis
• Minimize number of • Chemometrics
• Direct analysis
analysis • Unsupervised analysis
• Sample number and
amount reduc on

Data treatment
Sampling Bio-chemical
Hypothesis and
transport & conserva on analysis
interpreta on

Separa on
Extrac on Detec on
/clean-up

Traceability
• Minimize sample
• Minimize sample
treatment • Mul analyte
treatment
• Automa on and • Integrated techniques
• Waste reduc on
miniaturiza on • Waste reduc on
• In-vitro diges on
• Energy, waste, and • Reduced energy
• Bioavailability
reagent reduc on

Figure 1 Scheme of a typical foodomics study and its green possibilities.

4 Green Analytical Chemistry

depending on their food legislation and regulations. Nevertheless, all national specifications are harmonized by the Codex Alimen-
tarius Commission and World Trade Organization requirements, working together with public and private stakeholders at interna-
tional, national, and local levels to ensure quality and safety of foods (Food and Agriculture Organizations of the United Nations
Website, 2019).
Food and Agriculture Organizations of the United Nations (FAO-UN) have disseminated the so-called “needs assessment” for
food safety and quality (Food and Agriculture Organizations of the United Nations Website, 2019), where it is emphasized the idea
that technical tools must be developed to study technical and managerial aspects of food control to evaluate different aspects such
as: risk analysis, sampling and inspection, traceability, food safety and quality management, and specific quality schemes, among
others. In the same way, the European Commission recommends in the “Food safety: overview document” (European Commission,
2019) an integrated Food Safety policy to ensure a high level of protection of human health by the effective control of food produc-
tion to assess the compliance with EU standards, including also aspects like animal health, welfare, nutrition, and plant health.
Thus, a strict control is currently carried out by both, governments and private manufacturers, to assess the quality and safety of
food, involving a high amount of inspections and controls that require the analysis of a huge number of samples.
Foodomics is a new discipline, firstly defined by A. Cifuentes in 2009 as “a discipline that studies the Food and Nutrition
domains through the application of omics technologies” (Cifuentes, 2009). This concept comprises the control of all technological
processes and the effects on food, including aspects like food composition characterization and food nutritional value determina-
tion. Since its definition, Foodomics has had the responsibility to impulse food science research, especially in areas such as: the
improvement of food quality and food traceability, the fast resolution of food safety issues, and the impact evaluation of food
and its ingredients in human health (Cifuentes, 2018). In the last decades it has been observed the rising of high performance
analytical instruments (e.g., chromatography-mass spectrometry, capillary electrophoresis, high-resolution mass spectrometry,
advanced spectroscopy techniques, and nuclear magnetic resonance), chemometric tools, and novel methodologies applied to
food area, including the so-called massive -omic techniques such as genomics, proteomics, lipidomics, metabolomics, and nutri-
genomics (Bevilacqua et al., 2017). It typically targets answering to complex biological questions, related directly to the food or to
the effect of its ingestion in human health. These aspects are generally evaluated by massive untargeted approaches, with the
purpose of quantifying or semiquantifying all genes, proteins, or metabolites present in a sample. In this sense, advanced analytical
techniques play a fundamental role to identify and distinguish significant trends in the sample data that, after appropriate interpre-
tation, allows to know and elucidate the complexity of the food domain (Foodome) and, as a consequence, the concept of Green
Analytical Chemistry must be also taken into account (Gilbert-López et al., 2017). Concerning the potential impact of so many
determinations and analysis to be made on foods and the urgent need of fast response methodologies with no- or reduced-
deleterious effects on the environment, the greening of -omics platforms must be carried out considering basic operational aspects
related to direct analysis, when possible, sample preparation, when required, and separation techniques.

Direct Analysis: Dreams Come True

Among the analytical parameters to be determined in food to evaluate their quality characteristics and safety, many of these corre-
spond to major and minor components able to be determined directly in liquid and solid samples without any chemical pretreat-
ment. So, direct analysis looks as the greenest alternative in all the fields. However in the case of solid samples it must be taken into
consideration that not all the available techniques can quantify the chemical components in solids, being mandatory a previous
extraction of target analytes or a sample digestion to destroy the matrix.
To evaluate the potential alternatives for direct sample analysis we are forced to known from good to the best alternatives and,
as it is shown in Fig. 2, off-line analysis in the laboratory with a laser ablation (LA), laser induced breakdown (LIBs) or desorption
electrospray ionization (DESI) permits a highly sensitive and selective determination of many components of solid samples,
particularly based on the use of mass spectrometry (MS) or inductively coupled plasma optical emission (ICP-OES) or mass anal-
ysis of the ions generated (ICP-MS). Therefore, the use of the aforementioned techniques at-line or, in some cases in the field,
based on the use of portable instruments (Aramendia et al., 2019) could be more convenient alternatives than the use of bench
instruments.
On considering other techniques, as these base on direct atomic spectroscopic analysis of solids with damaging only some
sample aliquots, it is also possible to obtain sensitive data off-line or at-line. However, other instruments as energy dispersion
X-ray fluorescence (ED-XRF), Raman, MIR or NIR spectroscopy provide portable reduced cost instrumentation able to be used
in the field and also, in some cases, to obtain non-invasive or remote data. However, in these last cases, the sensitivity of determi-
nations could be reduced as compared with off-line alternatives.
Fig. 3 summarizes the green alternatives available in the literature for the direct analysis of solids in order to determine their
mineral and organic components. As it can be seen, image treatment seems the best alternative for determination of global param-
eters of foods (Dowlati et al., 2013), particular properties (Sanaeifar et al., 2016) or mineral elements (Zhou et al., 2019). Image
acquisition can be made by using the aforementioned simple photo cameras or smartphone or using hyperspectral cameras. In this
last case several applications have been reported especially in the last years (Liu et al., 2017).
Image treatment offers the greenest way for direct analysis of solid foods without manipulating the samples. However, it must be
taken into account that a previous chemometric modeling must be done and to do so some calibration samples must be analyzed
by rearguard methods because model prediction capability of the developed models cannot be higher than the quality of reference
 Green Analytical Chemistry 5

BEST

Remote sensing without

sample damage

Non-invasive analysis

In-field sampling
with portable
instruments

At-line analysis

Off-line analysis with

sample physical damage

GOOD

Figure 2 Hierarchical organization of green direct sample analysis.

MINERAL ELEMENTS

IMAGE X-Ray
Arc/Spark OES ETAAS Glow Discharge Laser Ablation LIBS
TREATMENT FLUORESCENCE

Direct Direct Direct Direct Slurry OES MS ICP-OES ICP-MS OES

determination determination solid
Sample Determination Determination
introduction

Direct Direct analysis on Direct determination Molecular

determination vials by ATR-FTIR MS fluorescence Raman

IMAGE
NIR Raman MIR DES LIBS
TREATMENT

ORGANIC COMPOUNDS
Figure 3 Green alternatives for direct solid sample analysis of mineral elements and organic components.

data used to create these calibrations. On the other hand, the applicability of the aforementioned strategies strongly depends on the
cost of instrumentation, being nowadays expensive the use of hyperspectral cameras and easy accessible the smartphones. Addition-
ally, the general use of image treatment as analytical tool will require, in a near future, the use of free software available through the
access from the cloud. Table 1 summarizes some of recent applications of image treatment based food analysis.
Raman and NIR methodologies (Li-Chan et al., 2010) are also non-invasive techniques which permit to obtain information
directly from the surface of samples, also covered by plastics or inside of glass vials; the same could be done by using ED-XRF
(Beckhoff et al., 2006). So, these methodologies are very useful. However, the sensitivity of these methods could cover the
percentage of target analyte levels with limit of detection values of ten per thousand. So it is difficult to use these approaches
when the interest is focused on trace analysis.
MIR direct analysis of foods involves the use of attenuated total reflection (ATR) measurements, and its use is very easy in the
case of liquid samples and permits the determination of many parameters from a single measurement (Moros et al., 2010). For solid
6 Green Analytical Chemistry

Table 1 Examples of applications in the food field of image processing methodologies

Technique/Camera Analyte/Parameter Sample Reference

Machine vision TVB-N, TBA and TVC Tilapia Shi et al. (2018)
Computer vision Quality indices Banana Sanaeifar et al. (2016)
Computer vision Color citrus index Citrus Vidal et al. (2013)
Machine vision Fish freshness Fish Dowlati et al. (2013)
Machine vision Honey characterization Honey Shafiee et al. (2013)
Machine vision Aflatoxin contaminated Pistachios Pearson et al. (1998)
Smartphone NIR spectrometer Fat, moisture and protein Salted minced meat Kartakoullis et al. (2019)
Smartphone Edible oil identification Vegetable oils Hakonen and Beves (2018)
Smartphone Oxygen Packed food Araque et al. (2018)
Smartphone Fat Cured meat products Cruz-Fernandez et al. (2017)
Smartphone Calories estimation Food Okamoto and Yanai (2016)
Smartphone Food recognition Food Kong and Tan. (2012)
Hyperspectral Cadmium Brown rice Zhou et al. (2019)
Hyperspectral Raman Maleic anhydride Starch powder Qin et al. (2018)
Hyperspectral 1000–2500 nm Fermentation index, polyphenol content, antioxidant Cocoa beans Caporaso et al. (2018)
activity
Hyperspectral 400–1000 nm Intramuscular fat content Boiled pork muscles Ma et al. (2018)
Hyperspectral 980–2500 nm Sucrose, caffeine, trigonelline Green coffee beans Caporaso et al. (2018)
Hyperspectral 980–2500 nm Protein content Wheat kernels Caporaso et al. (2018)
Hyperspectral 400–1800 nm Biogenic amines Cooked beef Yang et al. (2017a)
Hyperspectral 874–1734 nm Caffeine Coffee beans Zhang et al. (2017)
Hyperspectral 400–1000 nm TVB-N Cured meat Yang et al. (2017b)
Hyperspectral 400–1000 nm Hydroxyproline Chicken meat Xiong et al. (2015)
Hyperspectral 400–1000 nm Horsemeat adulteration Minced beef Kamruzzaman et al. (2015)
Hyperspectral (NIR) Melamine Milk powders Fu et al. (2014)
Hyperspectral (NIR) TVC Chicken breast fillets Feng and Sun (2013)
Hyperspectral 1100–2400 nm Ergot bodies Cereals Vermeulen et al. (2012)
Hyperspectral 900–1700 nm Omega-3 fatty acid Eggs Abdel-Nour and Ngadi (2011)
Hyperspectral 400–1000 nm Toxigenic fungi Maize Del Fiore et al. (2010)

Note: TVB-N, Total volatile basic nitrogen; TBA, Thiobarbituric acid; TVC, Total viable counts.

samples, obtaining reproducible signals is more critical due to the need of a good contact between sample and the optical element
of the ATR instrument; for this purpose existing instruments include a pressing holder to provide a constant pressure to the solid
sample. However, reproducibility of ATR measurements of solids is poorer than that obtained for liquid samples (Lu et al., 2017).
For mineral analysis arc/spark OES and ETAAS and glow discharge permits a direct analysis of sample by destroying an aliquot of
samples but with sensitivity levels which move from ppm to ppb concentration units. However, for these techniques, it is difficult to
move the instrument to the field and thus they remain at the off-line and at-line application levels.

Benefits of Greening Sample Preparation/Extraction

The definition of “modern” extraction technique could be something related to an environmentally safe, non-toxic, fast and auto-
mated, robust, highly sensitive and cost-efficient. In this context, “modern” extraction techniques have evolved from strong destruc-
tive processes, like the classical wet and dry ashing digestions, to soft techniques suitable to quantitatively extract the target analytes
from sample matrices. Soft treatments are those that imply analyte extraction without the complete dissolution of sample matrix.
They offer many advantages from the economic, environmental and fundamental analytical point of view as compared to hard
sample dissolution steps. Hard sample treatments involve high consumptions of energy, also increasing the potential interferences
during the measurement step due to the concomitant dissolution of interferences. Analyte extraction of organic compounds from
a solid sample, traditionally done by Soxhlet extraction, maceration or hydrodistillation, have been replaced by the considered
“modern” extraction techniques which include supercritical fluid extraction (SFE), pressurized liquid extraction (PLE), microwave
assisted extraction (MAE) and ultrasound assisted extraction (UAE).
The toxicity and volume of solvents used in the extraction step are the main aspects to be considered for greening the analytical
procedures, which should take into account a reduction of the energy required for the extraction. According to Jacotet-Navarro et al.
(2016) the main aims of green extraction technologies are related to: i) reduction of the extraction time, ii) efficient use of the
energy, iii) increase of the heat and mass transfer and iv) reduction of the equipment size.
The use of SFE with carbon dioxide, provides interesting environmental advantages, based on the low toxicity of CO2 and the fact
that its critical temperature is moderate (31.2  C). CO2 has been used for the extraction of bioactive compounds, as carotenoids,
 Green Analytical Chemistry 7

chlorophylls, phenolic compounds, vitamins, etc. (Bromberger Soquetta et al., 2018). However, the non-polar nature of CO2
limited the extraction yield of polar compounds, requiring the use of small amounts of organic modifiers like alcohols as polarity
modifiers. Other compound used as supercritical fluid is n-propane which has shown significant advantages, even compared to
carbon dioxide, such as high extraction yields, high solubility of non-polar compounds, short extraction times and reduced solvent
consumption (Hamdan et al., 2008). On the other hand, the supercritical conditions can imply a high energy consumption to
afford working temperature and pressure.
PLE involves the transfer of analytes from solid samples to liquid solvents at elevated temperatures and pressures. These condi-
tions produce a reduction in the surface tension of the solvent increasing the mass transfer of the analyte from the sample. The
solvents are selected based on the solubility of the desired analyte/s. The main advantages of the technique are translated to
fast extraction processes with reduced solvent consumption. Typical PLE extraction time ranges between 10 and 30 min at temper-
atures ranging between 60 and 200  C. The most commonly used solvent is ethanol, generally recognized as safe, readily available
at high purity and low cost and produced from biomass materials. The use of low volumes of innocuous or, at least, low toxicity
solvents must be one of the main factors to be considered. Using water as extraction solvent below its critical point and above
100  C and 20 atm is commonly known as pressurized hot water extraction (PHWE). It can be considered one of the greenest
extraction techniques. Acidified water (pH 2.5) has been also commonly used as solvent in PLE processes. Capello et al.
(2007) proposed a classification of organic solvents as a function of their green nature based on the application of the environ-
mental, health and safety (EHS) properties and the life cycle assessment (LCA) method of production, together with the potential
recycling and disposal of solvents. In this classification, organic solvents obtained from renewable sources, such as bioethanol or
vegetable oils, are considered potential green solvents from a LCA point of view; while heptane, hexane and diethyl ether are the
solvents of choice from the viewpoint of EHS. In any case, solvents that must be completely avoided are the substances of very high
concern (SVHC), such as dioxane, acetonitrile, formic and acetic acids, formaldehyde, tetrahydrofuran, chlorinated solvents, N-
methylpyrrolidinone, N,N-dimethyl formamide, and certain ethers of ethylene glycol (ECHA Candidate List of Substances of
Very High Concern for Authorization (2019)). Based on Green Analytical Chemistry principles, new solvents have been recently
designed to replace conventional organic solvents. Among the main alternatives, amphiphilic solvents (including surfactants, alco-
hols, and carboxylic acids), ionic liquids and derivatives, and deep eutectic solvents are the most explored candidates (Pacheco-
Fernández and Pino, 2019).
However, it should be taken into account that high efficient extraction apparatus, involving the use of high temperature and/or
pressure, require considerable energy consumption. In this sense, in recent days it has been proposal the use of hard cup espresso
machines to obtain fast extraction (in few seconds) at a temperature around 72  C and with a pressure of 1.9 MPa which offers a low
cost and efficient pressurized extraction tool (Armenta et al., 2016).
UAE is an effective extraction technique for a wide range of analytes from different samples. The main advantages of UAE are the
improvement of heat and mass transfer, easy to use, versatile, flexible and low cost. UAE is considered as a green alternative for the
extraction of bioactive compounds (Barba et al., 2016). On the other hand, MAE can be considered a green alternative extraction
technique because of it improves the heat and mass transfer of analytes from solid samples, reduces the extraction time due to the
use of high temperatures and pressure, and increases the extraction yield.
The use of solventless extraction procedures can be considered as the greenest approaches. They involve solid-phase microextrac-
tion (SPME), stir bar sorptive extraction (SBSE) or head-space (HS) direct injection. SPME can be done by HS extraction, exposing
the fiber in the vapor phase above a liquid or solid sample, or by direct immersion (DI-SPME) in a liquid sample. Desorption is
performed by heating the fiber, or by liquid desorption using an appropriate solvent. In any case, no or little solvent is needed, being
the technique easily coupled to chromatographic techniques. On the other hand, the main drawback is related to the lack of sensi-
tivity due to the small amount of sorbent coated onto the SPME fiber (Bicchi et al., 2002). This drawback can be avoided using SBSE,
where higher amounts of coating sorbents are employed. SBSE can be also performed by direct immersion or by HS exposition.
Additionally this technique increases the energy requirements.
Each extraction technique has its advantages and drawbacks and the selection of the most appropriate in each case depends on
several factors, mainly availability, acquisition and operational costs, simplicity, nature and amount of extraction solvents, energy
consumption, and obviously the green character of this step (Tobiszewski et al., 2009).
Moreover, in some cases, the extract may require an additional clean-up and/or preconcentration step prior the analytical deter-
mination. In those cases, there are several strategies proposed in the literature involving basically liquid–liquid and solid-phase
extraction (SPE) (García-Cañas et al., 2012). Miniaturization of liquid-liquid extraction procedures evolved toward single drop
extraction, dispersive liquid–liquid microextraction (DLLME) and hollow fiber liquid-phase microextraction (HF-LPME), among
others (Spietelun et al., 2014). These methods use only microvolumes of organic solvent and can be considered green preconcen-
tration/clean-up procedures.
The most used design in SPE is the polypropylene cartridge of 1 to 6 mL volume. Recent advances in SPE extraction formats
include disks, SPE pipette tip, and 96-well SPE microtiter plates among others. The idea of scaling down SPE has led to the devel-
opment of microextraction procedures, such as SPME, SBSE, stir rod sorptive extraction (SRSE), stir cake sorptive extraction (SCSE),
rotating disk sorptive extraction (RDSE), dispersive solid phase microextraction (DSPME), microextraction by packed sorbent
(MEPS), fiber-packed-needle microextraction (FNME), among others (Spietelun et al., 2013).
An alternative to SPE is the so called Quick, Easy, Cheap, Effective, Rugged and Safe extraction method (QuEChERS) (Rejczak
and Tuzimski, 2015), which involves the extraction of analytes from a homogenized sample using acetonitrile and salt solution and
the clean-up of the supernatant using a dispersive solid-phase extraction (DSPE) technique. QuEChERS can be considered as user-
8 Green Analytical Chemistry

friendly alternative to traditional liquid-liquid and solid-phase extractions. However, the use of acetonitrile, a SVHC considered by
the European Chemicals Agency, involves environmental risks.
A more in depth information about the use of the above mentioned green sample preparation/extraction methods in the Foo-
domics field can be found in different chapters within the book, devoted to the selection of environmentally friendly solvents, mini-
aturization of SPE and combination of assisted techniques, and green processes in Foodomics, including green solvents for
sustainable processes, Hansen solubility parameters (HSP) for green solvent selection, SFE, GXLs (Gas-expanded liquids), PLE,
PHWE, process integration and intensification and, finally, biorefinery.

Green Metrics Tools

In order to evaluate the greening degree of an analytical methodology, a quantitative tool is required to measure its impact on the
environment and it should be employed as an additional criterion to be taken into account in the selection of the most adequate
methodology in terms of environmental impact. To date, there is not yet a green metric tool that prevailed over others, and several
metric approaches have been proposed to evaluate the sustainability of analytical methods (see Fig. 4) (Armenta et al., 2019).
The first green metric approach was developed by the National Environmental Methods Index (NEMI) in 2002, as a database
that evaluated analytical methods according to their environmentally friendly performance based on four key qualitative aspects
in a pictogram: i) persistence, bioaccumulation, and toxicity of the employed reagents, ii) hazardous nature of processes and prod-
ucts, iii) reagent corrosiveness, and iv) generation of wastes (National Environmental Methods Index (NEMI), 2002). However,
NEMI concept showed a lack in the quantification of green aspects and did not include aspects related to energy consumption.
Thus, Raynie and Driven (2009) improved the NEMI criterion in 2009 by the incorporation of the energy consumption as a signif-
icant factor. This criterion was called Green Assessment Profile and it also considered health, safety and environmental hazards. de la
Guardia and Garrigues (2011) proposed a second modification of NEMI pictogram, incorporating a color scale (red, yellow and
green) as semiquantifiers to evaluate the environmental risks produced by reagents, waste generation, energy consumption, and
operator risks. Penalty points and Eco-scale concept (Galuszka et al., 2012) were proposed to quantify the Green Analytical Chem-
istry character of analytical methods. The use of Eco-scale assumes a totally green analytical method as 100 points and penalty
points are subtracted depending on the nature and consumption of reagents, energy use, and generated wastes. Green Motion
tool (Tony Phan et al., 2015), is based on the Eco-scale concept of penalty points but from a production point of view rather
than analytical one. In such strategy, penalty points are estimated considering the environmental, health, and safety impacts of
raw materials and results are represented in a spider-web graph. E-factor was proposed to evaluate the environmental impact of

Figure 4 Evolution of green metric tools pictograms over time.

 Green Analytical Chemistry 9

technological or industrial processes, calculated as the ratio of waste generated per weight of product (Tobiszewski et al., 2015).
Later in 2016, the Green Certificate was proposed (Armenta et al., 2016), a modification of the Eco-scale, incorporating mathemat-
ical equations to quantify the penalty points that are weighted considering the amount of employed resources. Moreover, green
certificate incorporates a global evaluation of the Green effectiveness of an analytical methodology using as colored letters (from
A to G). Finally, the most recent developed green metric tool is the Green Analytical Procedure Index (GAPI) (Płotka-Wasylka,
2018) that evaluates the green character of the entire analytical procedure, from sampling to the final determination, using
a five pentagram symbol to evaluate separately each step of the analytical procedure.
Other approaches have been proposed to evaluate the green character of particular methodologies like liquid chromatography,
such as i) the high performance liquid chromatography-environmental assessment tool (HPLC-EAT) that evaluates hazards related
to the nature and consumption of mobile phases in liquid chromatography (Gaber et al., 2011); and ii) the analytical method
volume intensity (AMVI) that evaluates the solvent consumption weighted by the chromatographic peaks of interest (Hartman
et al., 2011).
Examples of the application of the above mentioned Green Analytical Chemistry metrics are presented in last chapter of the
present book.
Additionally, different multivariate approaches have been proposed to identify and highlight environmental and health threats,
using computation tools like cluster analysis (Tobiszewski et al., 2013), self-organizing maps (Tobiszewski et al., 2014), multicri-
teria decision analysis (Al-Hazmi et al., 2016), or preference ranking organization method for enrichment evaluation (Promethee)
(Tobiszewski and Orlowski, 2015).
In summary, the use of green metrics approaches is contributing today to improve the selection of the most adequate method-
ology to be employed, from an environmental and sustainable point of view, to solve whatever analytical question. However, the
level of knowledge of the Green Analytical Chemistry concept among the scientific community is not comparable to procedures for
the quantification of the environmental impact or “greenness” of analytical procedures. Additional efforts should be done to avoid
that Green Analytical Chemistry remains as a philosophical concept, mainly, by the development of a simple, easily applicable and
universally recognized “greenness” evaluation approach suitable for method selection.

Future Trends and Efforts

In short, it can be concluded that Green Analytical Chemistry is a serious concern in the today analytical tasks and, particularly,
green methods should be developed and applied in the food field to preserve the safety of operators, the environmental health
and also to avoid sample contaminations and to assure a fast decision making.
We are absolutely convinced that advancements in high specifications instruments must be accompanied by the development of
vanguard fast screening methodologies based on high accurate reference values for a series of samples of the same type to those to be
analyzed; to do so, new alternatives as those based on image treatment will be in the spotlight in the following years.
On the other hand, efforts for screening the available methodologies and to search for new ones cannot forget that the greenest
method is not only the one with the lowest deleterious effects on the environment but also the one able to solve the analytical
problem with the required level of sensitivity, selectivity, precision and accuracy. Thus, a correct equilibrium between main analyt-
ical features and green parameters is absolutely required to select the appropriate methodologies.
On considering all above mentioned, it is time to move from theoretical considerations to the development and use of fully
validated green methods and to do it, additionally than checking the analytical features and method robustness, a complete eval-
uation of the analytical steps, the reagents and materials employed (also including their production), must be critically evaluated.
A final consideration concerns the complementarity between high sophisticated and high performance instrumental methods
and the fast screening ones in order to have a complete picture of the quality and state of foods and the correct evaluation of prob-
lems related to Foodomics.

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