Accepted Manuscript

Detection in UV-visible spectrophotometry: Detectors, detection systems, and

detection strategies

Marieta L.C. Passos, M. Lúcia M.F.S. Saraiva

PII: S0263-2241(18)31190-4
DOI: https://doi.org/10.1016/j.measurement.2018.12.045
Reference: MEASUR 6179

To appear in: Measurement

Received Date: 8 October 2018

Revised Date: 8 December 2018
Accepted Date: 11 December 2018

Please cite this article as: M.L.C. Passos, M.L.M.F. Saraiva, Detection in UV-visible spectrophotometry: Detectors,
detection systems, and detection strategies, Measurement (2018), doi: https://doi.org/10.1016/j.measurement.
2018.12.045

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Detection in UV-visible spectrophotometry: Detectors, detection systems,

and detection strategies

Marieta L. C. Passos*1, M. Lúcia M. F. S. Saraiva*2

LAQV, REQUIMTE, Department of Chemical Sciences, Laboratory of Applied Chemistry,

Faculty of Pharmacy, Porto University, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto,

Portugal

*Corresponding authors:
1
Name: Marieta L. C. Passos

E-mail address: [email protected]

Tel.: +351 220428643; Fax: +351 226093483.

2
Name: M. Lúcia M. F. S. Saraiva

E-mail address: [email protected]

Tel.: +351 220428674; Fax: +351 226093483.

Abstract

A compilation of the new developments in terms of detection, detections systems and

detection strategies in Ultraviolet-Visible (UV-Vis) spectrophotometry is presented and

discussed. It is shown that this evolution has promoted the use of UV-Vis spectrophotometry

as a simple, sensitive, reliable, and low cost technique, that allows the determination of very

low concentrations of compounds, and the use of very small amounts of samples. The

versatility, and the portability of the developed equipment and strategies, have also

contributed for the continuous use of UV-Vis spectrophotometry, over the years.

By the examples of its use in diverse fields such as agriculture, food, clinical, pharmaceutical,

and environmental is highly expectable that detection in UV-Vis spectrophotometry,

continues to evolve and to promote this kind of applications.

Keywords: UV-vis spectrophotometry; Organic compounds; Quantitative analysis;

Qualitative analysis; Detection; Detectors

 1. Introduction

The electromagnetic spectrum includes a wide range of energy frequencies, but the most

useful is in the range between >1019 (γ-rays) and 103 Hz (radiowaves). The visible region is a

small region of the spectrum and the visible light differ from the other type of radiation in the

frequency of energy of its photons. The absorption of light in the UV-Vis wavelength

(between ~180-800 nm) occurs frequently from many molecules such as a wide number of

organic molecules. The main energy changes happen at electronic level but can also happen at

vibrational quantum levels 1. n→σ* and σ → σ* transitions need more energy and are related

with absorption in the UV region and π →π* transition with absorption in the UV–Vis region.

The area of the molecule where the electronic transitions happen is called chromophore 2.

When the molecule is transparent in this region, derivatization can be used (Schiff base

formation, complexation reactions, enzymatic and other catalytic reactions, azodyes

derivatization, charge transfer complexation, ion-pair analysis, solid phase derivatization)3-6.

Using the derivatization, the sensitivity and sometimes the selectivity of the UV-Vis

methodology increase. The same happens when sample preconcentration/enrichment is used

(based on solvent or sorbent extraction or membrane separation) 7, 8. However, these are all

strategies to increase the possibility of an improved detection. During the recent past, a rapid

progress in the detection processes in UV-Vis spectrophotometry has been reported, in order

to improve the sensitivity, precision, and accuracy and at the same time to minimize the cost

and the time of analysis. However, most part of the published papers or book chapters are

focused on the basic principles of UV-Vis spectrophotometry 1, 2, 9, 10, on its applications in

specific fields such as the environment 11 or the pharmaceutical 3 or specific analytes such as

organic compounds or individual elements 12 and on the treatment of spectrophotometric

data13, 14. When the instrumentation is focused, the attention is divided between light sources,

monochromators, sample cells and detectors1. Even when the different detectors are compared
15
, it has been made in an abbreviated way. In the best of our knowledge there are not any

reference that compiled all the information about different kinds of detectors and, at the same

time, the detection system that have been developed using the referred detector associated

with other instrumentation, and the detection strategies that have been used in UV-Vis

spectrophotometry. Thus, in this paper, it is intended to highlight the different and more

actualized detectors, detection systems resulted from their development, and different

detection strategies used in UV-Vis spectrophotometry, showing a title of example some

applications in the determination of organic compounds. A flow diagram about the article

structure is presented in figure 1 in order to guide the readers throughout the text.

Please, insert here figure 1.

2. Principles of UV-Visible (UV-Vis) Spectrophotometry

Ultraviolet and Visible absorption spectrophotometry is the technique based on attenuation of

electromagnetic radiation measurement by an absorbing substance 9. This radiation, has a

spectral range approximately around 190-800 nm, which also differ in terms of energy ranges,

and type of excitation from other related regions (Table 1). This attenuation results from the

reflection, scattering, absorption or interferences. However, accurate measurements of the

attenuation can be done recording only the absorbance. Within some limits, the absorbance is

proportional to the concentration of the analyte to determine and to the distance of the light

when it passes through the sample during the irradiation. This relationship is called Beer’s

law and it is commonly written as A= ε x b x c, where A means absorbance, ε is the molar

absorbance coefficient (wavelength-dependent) in mol-1 L cm-1, b is the path length in cm and

c is the absorber concentration in mol L-1 9. This linear relationship can be influenced by

different factors such as the characteristics of the spectrophotometer, photodegradation of the

molecules, presence of scattering or absorbing interferences in the sample, fluorescent

compounds in the sample, interactions between the analyte and the solvent, and the pH 1, 9.

Please insert here Table 1.

3. Detectors

The function of a UV-Vis detector is to convert a light signal into an electric signal. Ideally, it

must respond in a wide wavelength range, respond with high sensitivity and with low noise,

have a linear response range, have a fast response, allow the miniaturization and a low

consumption of sample. In order to achieve an ideal detector, many detectors, that are

followed described, have been developed in the last few years.

3.1 Photomultipliers

Photomultiplier tubes (Figure 2) are composed by a photoemissive cathode, a series of

electrodes (dynodes) and an anode. The cathode when stricken by a photon, emits several

electrons that are accelerated towards the dynodes, since they are a release of many secondary

electrons. Those ones are accelerated to the next electrode where each one releases more

electrons. In the end, they are collected on the anode and consequently the final output is

further electronically amplified 16.

Please, insert here figure 2

The use of photomultipliers that are very sensitive, allow the detection of very low levels of

light, and high wavelength resolution since they permit the use of narrow slit widths. They

have been widely used for detections in UV and Visible regions.

3.2 Photodiode Detector Array (PDA)

The operation of a photodiode detector array is based on the fact that each diode in the array

is reverse-biased. Before the exposition of the detector to the light, the diodes are completed

charged through a transistor switch. When the light is reaching the PDA charge carriers will

be generated in the silicon. These carriers will neutralize the stored charges that have the

opposite polarity. Thus, this lost amount of charge is directly proportional to the light received

by the detector and it is measured during the recharge process of each diode, measuring the

amount of current need for the recharging 17.

PDA have the ability to monitor at all wavelengths. Thus, they have been widely used

as multichannel detectors. In addition, they can record a complete spectrum in a very short

period of time, since the movement of the diffractor does not affect the scan time.

Consequently, they are able to detect transient signals, such it happens in HPLC and flow

analysis 12. Additionally, PDA present some advantages such as the ability to quantify large

photon fluxes, high quantum efficiency, and the high height-to-width aspect ratio. Despite

these advantages, PDA present some drawbacks, such as a relevant dark current and a high

read noise 18. The development of charged-coupled devices was performed in order to resolve

these disadvantages.

3.3 Charged-Coupled Device (CCD)

The definition proposed by the Royal Society of Chemistry for the charged-coupled device is

a detector that uses a silicon chip to convert light into an electric signal. The Si chip absorbs a

photon and releases a single electron. The chip surface is covered by electrodes that hold the

electrons in an array of wells, or pixels. The charge built up corresponds to a pattern of

incident light 19. Thus, CCD present some advantages such as a low dark count rate, a low
read noise, and a high UV-Vis quantum efficiency 18, that make CCD excellent spectroscopic

detectors.

3.4 Paired Emitter Detector Diode (PEDD)

The Light-Emitting Diode (LED) consists of a p-n junction that emits a narrow band of light

frequencies when forward biased 20, 21. Since their development 22, LED have been used as an

interesting component in optical sensors 23-25, since they are inexpensive, allow the

miniaturization, the robustness and, a high efficiency. Despite their use started to be as light

sources, nowadays they have been applied in a reverse mode, as detectors 26, 27. When the

light is incident on the p-n junction, a current is generated. This current is proportional to the

light intensity 20, 21. Thus, since they can be used as a light source and detector, several

devices have been described based on Paired Emitter-Detector Diodes (PEDD) (Figure 3).

Please, insert here figure 3.

PEDD optical sensors have shown several advantages such as the low power consumption,

the reduced size, the sensitivity, the response in a large wavelength range with a high signal-

noise ratio. In addition, their output is a direct pulse-duration-modulated signal, that allows

the use of an expensive A/D converter unnecessary 28. Their advantages allow their use in

flow-through optical sensor used in chromatography 29, 30 and in flow analysis 31, 32, beyond

their use in different devices for simple absorbance measurements 33, 34.

4. Detection systems

Different detection systems resulted from the implementation of the different detectors

described above, have been developed to be used in UV-Vis spectrophotometry, in order to

increase the number of analyzed samples, the sensitivity, and selectivity, to decrease of the
reagents consumption, and to allow the automation when compared with the conventional

spectrophotometers.

4.1 Microplate reader

The use of a microplate optical reader results from the necessity to minimize the amount of

sample and to analyze a large number of samples in a short period of time. In this kind of

devices, the samples are placed in microplates pits instead of cells. The microplates usually

have between 6 and 1536 pits, but the most frequently used are the microplates with 96 pits.

These readers allow a thermostatic control and an orbital or a linear samples shaking before

the analysis. The measurements are made moving the plate relative to the fixed probe beam,

moving the latter relative to the plate or combining both strategies. More recently, it was

possible to measure simultaneously an entire strip using a PDA or a CCD line 35 (Figure 4).

Please, insert here figure 4.

Due to the capacity to analyse a large number of samples using small volumes of these

samples, the microplate reader have been used in different fields such as in environmental

analysis, for example in the determination of biochemical oxygen demand using 2,6-

dichlorophenolindophenol (DCIP) as a redox colour indicator 36. It has also been used in

clinical analysis, for example in the quantification of bacterial pathogens' virulence on the

insect model Galleria mellonella 37, in pharmaceutical field such as in the determination of

olmesartan medoxomil in tablets 38 or in food analysis, for example in the evaluation of

antioxidant compounds in fruits 39.

4.2 Charged-coupled device with optical fibers

Charged-coupled devices have been used in association with two optical fibers in order to

miniaturize the detection system. One of these fibers connect the light source to the cell

holder (where the sample is placed) and the other one connects this cell to the CCD detector.

This detection system is frequently used in association with flow systems for the

spectrophotometric quantification of organic compounds such as glucose 40 or for the

evaluation of enzymes activity such as the evaluation of peroxidase activity in fruits 41 or the

evaluation of glutathione reductase activity in order to predict the toxic effect of ionic liquids
42
.

4.3 Charged-coupled device with long path length cell

In order to improve the sensitivity of the method, a liquid waveguide capillary cell with a long

path length, have been coupled with CCD detectors, in flow analysis manifolds. This cell is

made of a fused silica tubing coating outside with a low refractive index polymer. The liquid

sample is guided through the hole of the capillary and consists in the core of the waveguide.

This kind of cell combines the use of small sample volumes (between 2.4 μL and 3 mL) with

an increase in the optical path length (between 10 and 500 cm) 43. This kind of strategy has

been frequently used for the evaluation of some elements such as bismuth in well waters 44,

cadmium in waters 45, and manganese in seawaters 46, but it has also been used to evaluate

organic compounds such as phenols in drinking waters 47, or Tc-99, an artificial compound

widely used in nuclear medicine for diagnostic tests48.

4.4 Devices for Point-of-Care Testing (PoCT)

The development of point-of-care testing has increased in the last decades. They intend to be

easier-to-use devices and allowing the performance of low-cost analysis in a non-laboratory

environment. There are two major categories of PoCT. The first one is composed of small

handheld devices, used either to qualitative or to quantitative analysis. Glucose biosensor

strips and lateral flow strips for the determination of cardiac markers and infectious pathogens

using immobilized antibodies are the most used technologies in this group. The second

category is composed of larger devices that result from the reduction in size and complexity

of laboratory instruments. In this category are included critical care analyzers, immunology

and small hematology analyzers 49. In order to promote, even more, the development of

patient-centered devices, the miniaturization of electronics and increasing computing power

have been taking place. Smart holograms 50, for example, has already been presented as a

future device.

4.5 Nanodrop

The NanoDrop technology is based on the development of a patented sample- retention

system that allows the use of very small volumes of sample (around 1μL). In these

microvolume spectrophotometers, the sample is placed between two optical surfaces, and the

surface tension of the sample allows the formation of a liquid column with a path length,

mechanically controlled (Figure 5). This control allows the use of a wide range of

concentrations, eliminating the need for dilutions. The surface tension of the sample promotes

the creation of its own container, dispensing the use of cuvettes, microcells or capillaries. This

kind of devices has been used for absorbance measurements in nucleic acids and proteins

analysis 51. It has been used, for example, to evaluate the integrity of genomic DNA isolated

from whole blood 52, for the quantification of cell-free DNA(cfDNA) in malignant melanoma

and prostate cancer patients 53, or for the analysis of gold nanoparticle-oligonucleotide

conjugate to detect the sequence of a lung cancer biomarker 54.

Please, insert here figure 5.

4.6 Information technology equipment (Digital cameras, video cameras and webcams, mobile

phones and scanners)

The use of information technology equipment (mobile phones, digital and video cameras,

scanners and, webcams), have been recently used as detection devices for colorimetric

chemistry. They have been applied using Microfluidic Papers based Analytical Devices

(μPADs), well plate reaction vessels and indicator papers. The use of these devices is simple,

the cost per analysis is low, and the portability allows their implementation in point of care

diagnosis and in-situ determinations in different fields 55. For example, a digital camera has

been used for the evaluation of 2,4,6-trinitrotoluene in soil extracts 56 or brilliant green in

wastewaters 57, a mobile phone for the determination of sulfadiazine and sulfasalazine in

pharmaceutical and veterinary formulations 58 and a webcam for the monitoring of

hydrochloric and phosphoric acids in aqueous solutions 59.

4.7 Integrated photodetectors

The integration of photodetectors in lab-on-chip has been reported for the development of

photonic biosensors. This integration allows an increment of the sensitivity, label-free and

real-time detection, promotes the automation and minimize the operator intervention. To

develop a photonic lab-on-chip device in the same platform it should be included the photonic

sensor, the valves, the pumps, the flow cell, the light source and the detector (a miniaturized

CCD camera or a PDA), the processing electronics and the firmware and software. This

integration could be considered monolithic if all the components are in the same chip or

hybrid if they are divided for different chips 60. This integration has allowed the development
of portable and easy-to-use devices for example for the evaluation of phenolic compounds 61,

antimicrobial metabolites 62, peptides 63 and drugs 64.

4.8 Spectrophotometer with integrating sphere

The combination of an integrating sphere with a UV-Vis spectrophotometer is useful for high

precision reflectance and scattered transmittance measurements. This allows the

characterization of solid samples, and photometric analysis of translucent, turbid and colloidal

samples. The analysis of this kind of samples in a conventional spectrophotometer results in

significant photometric errors and high variation between samples since the light is lost before

to reach the detector. The use of an integrating sphere avoids this behavior since it collects all

the light which has passed through the sample 65, 66. Thus, it is very useful, for example to

evaluate the optical properties of biological tissues such as the mucosa and submucosa of

human colon tissue 67, to characterize powders with encapsulated active pigments, such as the

yellow miraxanthin V and the violet betanidin, used in food applications 68, to measure the

ultraviolet/visible transmission of intraocular lenses 69, or to evaluate the concentration cells

such as cyanobacterium Anabaena variabilis 70.

particularly in quantitative pigment analyses of cell suspensions of photosynthetic

microorganisms.

5. Strategies for detection

5.1 Solid phase spectroscopy

The solid phase spectroscopy (SPS), is a photometric procedure, firstly described by

Yoshimura et al. 71 that is based on the immobilization of an analyte or a reaction product on a

solid phase. As solid phase, it can be used microbeads of non-polar or polymeric materials or

ion-exchange resins. After the obtained equilibrium between the sorbent active sites and the

target species, the beads are collected and placed in a cell with a few mL of a solution to be

measured as a suspension. The optical measurement is proportional to the analyte

concentration in the sample. The analyte retention on the solid support allows the analyte

preconcentration, resulting in a high sensitivity and selectivity of this procedure. However, to

obtain this enhanced sensitivity, it is crucial to use a high volume of sample (between 100-

1500 mL). Additionally, the consumption of solid support is also high since it is discarded at

every determination. However, when the batch concept is replaced by the implementation of a

SPS in flow-analysis systems, the solid phase starts to be regenerated after each determination,

performing successive measurements. Using this approach, the sorbent is placed inside the

flow cell and the retention, the preconcentration, and the detection of the analyte are

performed in the same place (the flow cell). The first time that SPS was performed in

association with flow analysis was in 1985 by Ruzicka et. al. 72. Since then, it has been

reported the association of SPS with different kinds of flow systems (flow injection analysis,

sequential injection analysis, multi-syringe flow injection, and multicommutation) 40, 73. This

promotes the association of different features such as the miniaturization with a subsequent

reagent and sample consumption and waste generation, the automation, the re-use of the solid

phase (except in bead injection approach), the versatility, the high sensitivity and selectivity 74.
5.2 Online analysis

Nowadays, the flow analysis techniques, such as Flow Injection Analysis (FIA) 75, Sequential

Injection Analysis (SIA) 76, Multicommutation Flow Injection Analysis (MCFIA) 77,

Multisyringe Flow Injection Analysis (MSFIA) 78, and Multi-Pumping Flow System (MPFS)
79
have shown to be great tools in the laboratories. This kind of flow systems present several

advantages comparing with batch procedures, such as, the increment of sample throughput,

allow the reduction of sample and reagents consumption and consequently the reduction in

the cost per analysis and the waste generation, allow the automation, reducing the operator

intervention, the errors and the time spent per analysis, allow online sample pretreatment

(including dialysis, gas diffusion, digestion, separation, dilution or preconcentration). The

miniaturization of these flow systems resulted in Lab-On-Valve (LOV) 80 systems as well as

microfluidic devices that allow the development of new microflow analytical systems. In

addition, these systems can integrate different kinds of detectors, improving the selectivity

and the sensitivity 81, 82. However, UV-Vis spectrophotometry is the most common choice,

frequently associated with chemical derivatization, due to the simplicity and adequate

selectivity 82. In some cases, some strategies can be used to decrease the detection limit such

as using long pathlength cells, as it happens in the determination of pesticides in milk 83 or

antibiotics in honey 84.

5.3 Hyphenated techniques

Hyphenation consists in the combination between a spectroscopic detection with a separation

technique with the objective to obtain structural information about the analytes 85, 86. Liquid

Chromatography-Mass Spectrometry (LC-MS), and Gas Chromatography-Mass Spectrometry

(GC-MS) are hyphenated techniques frequently used 86. However, sometimes, complementary

information is required. Thus, LC and GC are hyphenated with other kinds of detection,
including the use of a PDA. The LC-UV hyphenation has been used for example, in the

pharmaceutical, biomedical, environmental and food analysis 86. The UV spectrophotometry

is rarely used as CG detection since there are a reduced number of GC amenable compound

that absorb in the UV range, however, this number is enhanced in the Vacuum Ultraviolet

(VUV) region (80-190 nm) 87 (Figure 6).

Please, insert here figure 6.

The VUV spectrophotometer was developed in order to overcome some limitations presented

by other detectors and it performs the deconvolution of co-eluting analytes, allowing

qualitative and quantitative analysis 87. The CG-VUV hyphenation has been used in different

applications, such as, in the multiclass pesticide determination 88, in natural gas

characterization 89, and in fatty acid methyl esters (including saturated, monounsaturated and

polyunsaturated fatty acids) analysis 90.

5.4 Hypernation and Extended-Hypernation Techniques

A combination of different hyphenated techniques in the same setup is called hypernation and

sometimes is an option for solving some analytical problems 86. The hypernation use is

advantageous in terms of runtime, data correlation, and sample size but it presents the

disadvantage of the use of diverse and expensive spectrophotometers 86. Recently, it has been

developed hypernation techniques that combine UV, Inductively Coupled Plasma-MS (ICP-

MS), MS, Nuclear Magnetic Resonance (NMR), and Fourier Transform Infrared (FTIR),

detectors with LC-based operations. The hypernation LC/UV/MS has been referred as useful

to distinguish several sub-classes of flavonoids 91 and in the investigation of phytochemical

products, when additional information is crucial 92, 93. The combination of VUV detection
with a GC x GC has been reported for the measurements of volatile organic compounds in

bread gas 94 and for hydrocarbon fuels analysis 95, in order to complement the selectivity of a

mass spectrometer.

6. Conclusions

A high evolution in the UV-Vis detection has happened in the last years. The development of

different kinds of detectors have been occurring in order to increase the detection capacity and

to minimize some particular drawbacks. Coupling these detectors with other instrumentation,

such as optical fibers or long path length cells, some new detection systems have appeared.

New strategies of detection, such as the solid phase spectroscopy or the hyphenation with

some other techniques were also described in the last years.

All the detectors, detection systems or detection strategies were developed in order to promote

simple and low cost analysis, the decrease in the detection limits, and the increase in the

sensitivity of different methodologies of analysis. Thus, it is not possible to elect the best

detector, detection system or detection strategy, since each one has particular characteristics

that can be useful in different situations according for example with the analyte, with analyte

concentration in the sample, the reaction involved in the determination, the available sample

volume, the sample matrix, or the necessity of portability.

In the future, it is highly expectable that detection in UV-Vis spectrophotometry continues to

evolve in order to keep up with the necessity to determine very low concentrations of

compounds and to promote the quantification in samples available in very small quantities.

The portability, the versatility, and the simplicity of the equipment, and its mode of use will

always be present during the development of new detectors and new strategies of detection.

Conflicts of interest
There are no conflicts of interest to declare.

Acknowledgements

This work received financial support from the European Union (FEDER funds

POCI/01/0145/FEDER/007265) and National Funds (FCT/MEC, Fundação para a Ciência e

Tecnologia and Ministério da Educação e Ciência) under the Partnership Agreement PT2020

UID/QUI/50006/2013. Also by FEDER Funds through the Operational Competitiveness

Factors Program - COMPETE and by National Funds through FCT - within the scope of the

project POCI-01-0145-FEDER-030163. Marieta Passos thanks FCT for the financial support.
References

1. L. Sommer, in STUDIES IN ANALYTICAL CHEMISTRY, eds. E. Pungor, W.

Simon, H. Malissa and J. Inczedy, Elsevier, Amsterdam, 1989.

2. R. J. Anderson, D. J. Bendell and P. W. Groundwater, in Organic Spectroscopic

Analysis, eds. R. J. Anderson, D. J. Bendell and P. W. Groundwater, The Royal

Society of Chemistry, 2004, vol. 22, pp. 7-23.

3. O. Adegoke, Chemical derivatization methodologies for UV-visible

spectrophotometric determination of pharmaceuticals, International Journal of

Pharmaceutical Sciences Review and Research, 2012, 14, 6-24.

4. G. S. Deokar, I. S. Pagare, A. M. Naseem, S. Kshirsagar and L. Shindhe,

Chemical Derivatization UV Spectrophotometric Method for Detection of P-

Aminophenol and Energy of Activation Approach to Set Degradation Protocol

for Forced Degradation Studies, International Journal of Pharma Research

review, 2016, 5, 1-12.

5. C. P. Babalola, I. Oluwalana, O. A. Kotila, O. A. Adegoke, Y. T. Kolade and S.

J. Ameh, A Novel Derivatization Ultraviolet Spectrophotometric Method for the

Determination of Dihydroartemisinin using p-Nitroaniline, Tropical Journal of

Pharmaceutical Research, 2014, 13, 127-133.

6. T. Catrinck, A. Dias, M. C. S. Aguiar, F. O. Silverio, P. H. Fidencio and G. P.

Pinho, A Simple and Efficient Method for Derivatization of Glyphosate and

AMPA Using 9-Fluorenylmethyl Chloroformate and Spectrophotometric

Analysis, Journal of the Brazilian Chemical Society, 2014, 25, 1194-1199.

7. P. L. Buldini, L. Ricci and J. L. Sharma, Recent applications of sample

preparation techniques in food analysis, Journal of Chromatography A, 2002,

975, 47-70.
8. M. D. M. Abadi, N. Ashraf, M. Chamsaz and F. Shemirani, An overview of

liquid phase microextraction approaches combined with UV-Vis

spectrophotometry, Talanta, 2012, 99, 1-12.

9. B. M. Tissue, in Characterization of Materials, ed. E. N. Kaufmann, John Wiley

& Sons, Inc., 2012, DOI: doi:10.1002/0471266965.com059.pub2.

10. S. Kumar, in Organic chemistry, 2008.

11. C. B. Ojeda and F. S. Rojas, Process Analytical Chemistry: Applications of

Ultraviolet/Visible Spectrometry in Environmental Analysis: An Overview,

Applied Spectroscopy Reviews, 2009, 44, 245-265.

12. R. Lobinski and Z. Marczenko, Recent advances in ultraviolet-visible

spectrophotometry, Critical Reviews in Analytical Chemistry, 1992, 23, 55-111.

13. F. S. Rojas and C. B. Ojeda, Recent development in derivative ultraviolet/visible

absorption spectrophotometry: 2004-2008, Analytica Chimica Acta, 2009, 635,

22-44.

14. C. B. Ojeda and F. S. Rojas, Recent applications in derivative ultraviolet/visible

absorption spectrophotometry: 2009-2011 A review, Microchemical Journal,

2013, 106, 1-16.

15. W. E. L. Grossman, A comparison of optical-detectors for the visible and

unltraviolet, Journal of Chemical Education, 1989, 66, 697-700.

16. G. D. Christian, P. K. Dasgupta and S. K. A., Analytical Chemistry, Wiley,

seventh edition edn., 2014.

17. W. E. L. Grossman, A comparison of optical detectors for the visible and

ultraviolet, Journal of Chemical Education, 1989, 66, 697.

18. J. V. Sweedler, R. D. Jalkian and M. B. Denton, A Linear Charge-Coupled

Device Detector System for Spectroscopy, Applied Spectroscopy, 1989, 43, 953-

962.

19. R. S. Chemistry, Charge-coupled-device detector,

http://www.rsc.org/publishing/journals/prospect/ontology.asp?id=CMO:000224

5&MSID=B819455F, (accessed 09/02/2018, 2018).

20. L. Tymecki and R. Koncki, Simplified paired-emitter-detector-diodes-based

photometry with improved sensitivity, Analytica Chimica Acta, 2009, 639, 73-

77.

21. L. Tumecki, L. Brodacka, B. Rozum and R. Koncki, UV-PEDD photometry

dedicated for bioanalytical uses, Analyst, 2009, 134, 1333-1337.

22. N. Holonyak and S. F. Bevacqua, Coherent (visible) light emission from

GA(AS1-XPX) junctions, Applied Physics Letters, 1962, 1, 82-83.

23. P. K. Dasgupta, I. Y. Eom, K. J. Morris and J. Z. Li, Light emitting diode-based

detectors absorbance, fluorescence and spectroelectrochemical measurements in

a planar flow-through cell, Analytica Chimica Acta, 2003, 500, 337-364.

24. J. Kovac, L. Peternai and O. Lengyel, Advanced light emitting diodes structures

for optoelectronic applications, Thin Solid Films, 2003, 433, 22-26.

25. P. K. Dasgupta, H. S. Bellamy, H. H. Liu, J. L. Lopez, E. L. Loree, K. Morris, K.

Petersen and K. A. Mir, Light-emitting diode based flow-through optical-

absortion detectors, Talanta, 1993, 40, 53-74.

26. F. M. Mims, Sun photometer with light-emitting-diodes as spectrally selective

detectors, Applied Optics, 1992, 31, 6965-6967.

27. E. Miyazaki, S. Itami and T. Araki, Using a light-emitting diode as a high-speed,

wavelength selective photodetector, Review of Scientific Instruments, 1998, 69,

3751-3754.

28. M. O'Toole and D. Diamond, Absorbance based light emitting diode optical

sensors and sensing devices, Sensors, 2008, 8, 2453-2479.

29. L. Barron, M. O'Toole, D. Diamond, P. N. Nesterenko and B. Paull, Separation

of transition metals on a poly-iminodiacetic acid grafted polymeric resin column

with post-column reaction detection utilising a paired emitter-detector diode

system, Journal of Chromatography A, 2008, 1213, 31-36.

30. M. O. Toole, L. Barron, R. Shepherd, B. Paull, P. Nesterenko and D. Diamond,

Paired emitter-detector diode detection with dual wavelength monitoring for

enhanced sensitivity to transition metals in ion chromatography with post-

column reaction, Analyst, 2009, 134, 124-130.

31. M. O'Toole, K. T. Lau and D. Diamond, Photometric detection in flow analysis

systems using integrated PEDDs, Talanta, 2005, 66, 1340-1344.

32. M. O'Toole, K. T. Lau, R. Shepherd, C. Slater and D. Diamond, Determination

of phosphate using a highly sensitive paired emitter-detector diode photometric

flow detector, Analytica Chimica Acta, 2007, 597, 290-294.

33. K. T. Lau, E. McHugh, S. Baldwin and D. Diamond, Paired emitter-detector

light emitting diodes for the measurement of lead(II) and cadmium(II), Analytica

Chimica Acta, 2006, 569, 221-226.

34. K. T. Lau, W. S. Yerazunis, R. L. Shepherd and D. Diamond, Quantitative

colorimetric analysis of dye mixtures using an optical photometer based on LED

array, Sensors and Actuators B-Chemical, 2006, 114, 819-825.

35. A. D. Levin, Use of Microplate Optical Readers in Biomedical Measurements,

Measurement Techniques, 2005, 48, 293-298.

36. N. Yoshida, S. J. McNiven, T. Morita, H. Nakamura and I. Karube, A simple,

multiple simultaneous spectrophotometric method for bod determination using

DCIP as the redox color indicator, Analytical Letters, 2002, 35, 1541-1549.

37. N. Parthuisot, J. Rouquette and J. B. Ferdy, A high-throughput technique to

quantify bacterial pathogens' virulence on the insect model Galleria mellonella,

Journal of Microbiological Methods, 2018, 152, 69-72.

38. I. A. Darwish, T. A. Wani, N. Y. Khalil and H. M. Abdel-Rahman, High

throughput microwell spectrophotometric assay for olmesartan medoxomil in

tablets based on its charge-transfer reaction with DDQ, Acta Pharmaceutica,

2014, 64, 63-75.

39. F. Saidani, R. Gimeneze, C. Aubert, G. Chalot, J. A. Betran and Y. Gogorcena,

Phenolic, sugar and acid profiles and the antioxidant composition in the peel and

pulp of peach fruits, Journal of Food Composition and Analysis, 2017, 62, 126-

133.

40. M. L. Passos, M. L. Saraiva and J. L. Lima, A thionine-based reversible redox

sensor in a sequential injection system, Analytica Chimica Acta, 2010, 668, 41-

46.

41. D. Costa, M. L. C. Passos, A. M. O. Azevedo and M. Saraiva, Automatic

evaluation of peroxidase activity using different substrates under a micro

sequential injection analysis/lab-on-valve (mu SIA-LOV) format,

Microchemical Journal, 2017, 134, 98-103.

42. E. Cunha, M. L. C. Passos, P. Pinto and M. Saraiva, Automated evaluation of

the inhibition of glutathione reductase activity: application to the prediction of

ionic liquids' toxicity, Rsc Advances, 2015, 5, 78971-78978.

43. W. P. Instruments, Liquid Waveguide Capillary Cell, 100 cm pathlength,

https://www.wpiinc.com/products/spectroscopy-and-optics/lwcc-3100-liquid-

waveguide-capillary-cell-100-cm-pathlength/, (accessed 30/01/2018, 2018).

44. C. Calderilla, J. Avivar, L. O. Leal and V. Cerda, Multivariate optimisation of a

rapid and simple automated method for bismuth determination in well water

samples exploiting long path length spectrophotometry, International Journal of

Environmental Analytical Chemistry, 2016, 96, 653-666.

45. T. D. Magalhaes and B. F. Reis, A novel multicommuted flow analysis strategy

for the spectrophotometric determination of cadmium in water at mu g L-1

levels without using a preconcentration step, Analytical Methods, 2018, 10, 900-

909.

46. S. C. Feng, D. X. Yuan, Y. M. Huang, K. N. Lin, Y. Zhu and J. Ma, A catalytic

spectrophotometric method for determination of nanomolar manganese in

seawater using reverse flow injection analysis and a long path length liquid

waveguide capillary cell, Talanta, 2018, 178, 577-582.

47. B. Horstkotte, F. Maya, C. M. Duarte and V. Cerdà, Determination of ppb-level

phenol index using in-syringe dispersive liquid-liquid microextraction and liquid

waveguide capillary cell spectrophotometry, Microchimica Acta, 2012, 179, 91-

98.

48. M. Villar, A. Borras, J. Avivar, F. Vega, V. Cerda and L. Ferrer, Fully

Automated System for Tc-99 Monitoring in Hospital and Urban Residues: A

 Simple Approach to Waste Management, Analytical Chemistry, 2017, 89, 5858-

5864.

49. A. St John and C. P. Price, Existing and Emerging Technologies for Point-of-

Care Testing, Clinical Biochemical Reviews, 2014, 35, 155-167.

50. S. Kabilan, A. J Marshall, A. Horgan, C. D Creasey, S. Kew, K. E S Dean, S. F

Terrell and L. J Affleck, "Smart" Holograms – A Novel Diagnostics Platform,

2006.

51. P. Desjardins, Micro-volume spectroscopy: thinking outside the cuvette, Nature

Methods, 2007, AN26-AN27.

52. W. C. Chen, R. Kerr, A. May, B. Ndlovu, A. Sobalisa, S. T. Duze, L. Joseph, C.

G. Mathew and C. B. de Villiers, The Integrity and Yield of Genomic DNA

Isolated from Whole Blood Following Long-Term Storage at-30 degrees C,

Biopreservation and Biobanking, 2018, 16, 106-113.

53. G. Ponti, M. Maccaferri, M. Manfredini, S. Kaleci, M. Mandrioli, G. Pellacani,

T. Ozben, R. Depenni, G. Bianchi, G. M. Pirola and A. Tomasi, The value of

fluorimetry (Qubit) and spectrophotometry (NanoDrop) in the quantification of

cell-free DNA (cfDNA) in malignant melanoma and prostate cancer patients,

Clinica Chimica Acta, 2018, 479, 14-19.

54. H. Daraee, M. Pourhassanmoghadam, A. Akbarzadeh, N. Zarghami and M.

Rahmati-Yamchi, Gold nanoparticle-oligonucleotide conjugate to detect the

sequence of lung cancer biomarker, Artificial Cells Nanomedicine and

Biotechnology, 2016, 44, 1417-1423.

55. K. Grudpan, S. D. Kolev, S. Lapanantnopakhun, I. D. McKelvie and W.

Wongwilai, Applications of everyday IT and communications devices in modern

analytical chemistry: A review, Talanta, 2015, 136, 84-94.

56. A. Choodum, P. Kanatharana, W. Wongniramaikul and N. NicDaeid, Rapid

quantitative colourimetric tests for trinitrotoluene (TNT) in soil, Forensic

Science International, 2012, 222, 340-345.

57. S. Damirchi, M. A. K. Khooni, T. Heidari, Z. Es'haghi and M. Chamsaz, A

comparison between digital camera and spectrophotometer for sensitive and

selective kinetic determination of brilliant green in wastewaters, Spectrochimica

Acta Part a-Molecular and Biomolecular Spectroscopy, 2019, 206, 232-239.

58. S. A. Errayess, L. Idrissi and A. Amine, Smartphone-based colorimetric

determination of sulfadiazine and sulfasalazine in pharmaceutical and veterinary

formulations, Instrumentation Science & Technology, 2018, 46, 656-675.

59. E. D. Gaiao, V. L. Martins, W. D. Lyra, L. F. de Almeida, E. C. da Silva and M.

C. U. Araujo, Digital image-based titrations, Analytica Chimica Acta, 2006, 570,

283-290.

60. M. C. Estevez, M. Alvarez and L. M. Lechuga, Integrated optical devices for

lab-on-a-chip biosensing applications, Laser & Photonics Reviews, 2012, 6, 463-

487.

61. S. Wakida, K. Fujimoto, H. Nagai, T. Miyado, Y. Shibutani and S. Takeda, On-

chip micellar electrokinetic chromatographic separation of phenolic chemicals in

waters, Journal of Chromatography A, 2006, 1109, 179-182.

62. E. Guihen and J. D. Glennon, Rapid separation of antimicrobial metabolites by

microchip electrophoresis with UV linear imaging detection, Journal of

Chromatography A, 2005, 1071, 223-228.

63. R. Jindal and S. M. Cramer, On-chip electrochromatography using sol-gel

immobilized stationary phase with UV absorbance detection, Journal of

Chromatography A, 2004, 1044, 277-285.

64. M. Ludwig, F. Kohler and D. Belder, High-speed chiral separations on a

microchip with UV detection, Electrophoresis, 2003, 24, 3233-3238.

65. G. Bain,Integrating Sphere Diffuse Reflectance Technology for use with UV-

Visible Spectrophotometry - Technical note 51450, Journal, 2007.

66. PerkinElmer,Applications and Use of Integrating Spheres With the LAMBDA

650 and 850 UV/Vis and LAMBDA 950 UV/Vis/NIR Spectrophotometers -

Application note, Journal.

67. A. N. Bashkatov, E. A. Genina, V. I. Kochubey, V. S. Rubtsov, E. A.

Kolesnikova and V. V. Tuchin, Optical properties of human colon tissues in the

350-2500 nm spectral range, Quantum Electronics, 2014, 44, 779-784.

68. F. Gandia-Herrero, J. Cabanes, J. Escribano, F. Garcia-Carmona and M.

Jimenez-Atienzar, Encapsulation of the Most Potent Antioxidant Betalains in

Edible Matrixes as Powders of Different Colors, Journal of Agricultural and

Food Chemistry, 2013, 61, 4294-4302.

69. A. Akinay, M. D. Ong, M. Choi and M. Karakelle, Measuring ultraviolet-visible

light transmission of intraocular lenses: double-beam mode versus integrating-

sphere mode, Journal of Biomedical Optics, 2012, 17, 7.

70. M. N. Merzlyak and K. R. Naqvi, On recording the true absorption spectrum and

the scattering spectrum of a turbid sample: application to cell suspensions of the

cyanobacterium Anabaena variabilis, Journal of Photochemistry and

Photobiology B-Biology, 2000, 58, 123-129.

71. K. Yoshimura, H. Waki and S. Ohashi, Ion-exchanger colorimetry.1. Micro-

determination of chromium, iron, copper and cobalt in water, Talanta, 1976, 23,

449-454.
72. J. Ruzicka and E. H. Hansen, Optosensing at active surfaces - A new detection

principle in flow-injection analysis, Analytica Chimica Acta, 1985, 173, 3-21.

73. E. J. Llorent-Martinez, A. Dominguez-Vidal, P. Ortega-Barrales, M. de la

Guardia and A. Molina-Diaz, Implementation of multicommutation principle

with flow-through multioptosensors, Analytica Chimica Acta, 2005, 545, 113-

118.

74. A. Molina-Díaz, J. F. García-Reyes and B. Gilbert-López, Solid-phase

spectroscopy from the point of view of green analytical chemistry, TrAC Trends

in Analytical Chemistry, 2010, 29, 654-666.

75. J. Ruzicka and E. H. Hansen, Flow injection analysis.1. New concept of fast

continuous-flow analysis, Analytica Chimica Acta, 1975, 78, 145-157.

76. J. Ruzicka and G. D. Marshall, Sequential injection - A new concept for

chemical sensors, process analysis and laboratory assays, Analytica Chimica

Acta, 1990, 237, 329-343.

77. B. F. Reis, M. F. Gine, E. A. G. Zagatto, J. Lima and R. A. Lapa,

Multicommutation in flow-analysis .1. Binary sampling - Concepts,

instrumentation and spectrophotometric determination of iron in plant digests,

Analytica Chimica Acta, 1994, 293, 129-138.

78. F. Albertus, B. Horstkotte, A. Cladera and V. Cerda, A robust multisyringe

system for process flow analysis - Part I. On-line dilution and single point

titration of protolytes, Analyst, 1999, 124, 1373-1381.

79. R. A. S. Lapa, J. Lima, B. F. Reis, J. L. M. Santos and E. A. G. Zagatto, Multi-

pumping in flow analysis: concepts, instrumentation, potentialities, Analytica

Chimica Acta, 2002, 466, 125-132.

80. J. Ruzicka, Lab-on-valve: universal microflow analyzer based on sequential and

bead injection, Analyst, 2000, 125, 1053-1060.

81. V. Cerda, L. Ferrer, L. A. Portugal, C. T. de Souza and S. L. C. Ferreira,

Multisyringe flow injection analysis in spectroanalytical techniques - A review,

Trac-Trends in Analytical Chemistry, 2018, 98, 1-18.

82. M. K. Sasaki, F. R. P. Rocha, A. D. Batista and D. L. Rocha, Flow-based food

analysis: an overview of recent contributions, Analytical Methods, 2017, 9,

6313-6334.

83. M. P. Rodriguez, B. F. da Silva, H. R. Pezza and L. Pezza, A greener flow

injection method based on a LWCC for the screening of tetracycline antibiotics

in bovine milk samples, Analytical Methods, 2016, 8, 5262-5271.

84. T. A. Catelani, I. V. Toth, J. Lima, L. Pezza and H. R. Pezza, A simple and rapid

screening method for sulfonamides in honey using a flow injection system

coupled to a liquid waveguide capillary cell, Talanta, 2014, 121, 281-287.

85. U. A. T. Brinkman, Hyphenation:Hype and fascination, Elsevier Science,

Amsterdam, 1999.

86. I. D. Wilson and U. A. T. Brinkman, Hyphenation and hypernation - The

practice and prospects of multiple hyphenation, Journal of Chromatography A,

2003, 1000, 325-356.

87. I. C. Santos and K. A. Schug, Recent advances and applications of gas

chromatography vacuum ultraviolet spectroscopy, Journal of Separation Science,

2017, 40, 138-151.

88. H. Fan, J. Smuts, P. Walsh, D. Harrison and K. A. Schug, Gas chromatography–

vacuum ultraviolet spectroscopy for multiclass pesticide identification, Journal

of Chromatography A, 2015, 1389, 120-127.

89. L. Bai, J. Smuts, P. Walsh, H. Fan, Z. Hildenbrand, D. Wong, D. Wetz and K. A.

Schug, Permanent gas analysis using gas chromatography with vacuum

ultraviolet detection, Journal of Chromatography A, 2015, 1388, 244-250.

90. H. Fan, J. Smuts, L. Bai, P. Walsh, D. W. Armstrong and K. A. Schug, Gas

chromatography-vacuum ultraviolet spectroscopy for analysis of fatty acid

methyl esters, Food Chem, 2016, 194, 265-271.

91. H. M. Merken and G. R. Beecher, Measurement of Food Flavonoids by High-

Performance Liquid Chromatography: A Review, Journal of Agricultural and

Food Chemistry, 2000, 48, 577-599.

92. J.-L. Wolfender, K. Ndjoko and K. Hostettmann, Liquid chromatography with

ultraviolet absorbance–mass spectrometric detection and with nuclear magnetic

resonance spectrometry: a powerful combination for the on-line structural

investigation of plant metabolites, Journal of Chromatography A, 2003, 1000,

437-455.

93. J. L. Wolfender, K. Ndjoko and K. Hostettmann, The potential of LC-NMR in

phytochemical analysis, Phytochemical Analysis, 2001, 12, 2-22.

94. B. Gruber, T. Groeger, D. Harrison and R. Zimmermann, Vacuum ultraviolet

absorption spectroscopy in combination with comprehensive two-dimensional

gas chromatography for the monitoring of volatile organic compounds in breath

gas: A feasibility study, Journal of Chromatography A, 2016, 1464, 141-146.

95. T. Groger, B. Gruber, D. Harrison, M. Saraji-Bozorgzad, M. Mthembu, A.

Sutherland and R. Zimmermann, A Vacuum Ultraviolet Absorption Array

Spectrometer as a Selective Detector for Comprehensive Two-Dimensional Gas

Chromatography: Concept and First Results, Analytical Chemistry, 2016, 88,

3031-3039.
96. N. Jespersen, in Comprehensive Analytical Chemistry, eds. S. Ahuja and N.

Jespersen, Elsevier, 2006, vol. 47, pp. 111-155.

97. T. Scientific, https://www.thermofisher.com/pt/en/home/industrial/spectroscopy-

elemental-isotope-analysis/molecular-spectroscopy/ultraviolet-visible-visible-

spectrophotometry-uv-vis-vis/uv-vis-vis-instruments/nanodrop-microvolume-

spectrophotometers/nanodrop-products-guide/nanodrop-how-it-works.html).

98. K. A. Schug, I. Sawicki, D. D. Carlton, H. Fan, H. M. McNair, J. P. Nimmo, P.

Kroll, J. Smuts, P. Walsh and D. Harrison, Vacuum Ultraviolet Detector for Gas

Chromatography, Analytical Chemistry, 2014, 86, 8329-8335.

Figure Captions

Figure 1: Flow diagram representing the structure of the paper.

Figure 2: A schematic diagram of a photomultiplier tube. (Reprint from Jespersen 96

with permission from the Elsevier. Copyright 2006.)

Figure 3: A schematic of the integrated PEDD flow analysis device used for

colorimetric detection. (Reprint from O’Toole et. al. 28 under the Creative Commons

Attribution Licence (CC BY 3.0))

Figure 4: Simultaneous data readout from a microplate strip: 1) fiber light guide; 2, 4)

microlenses; 3) plate strip; 5) measurement channel photodiodes; 6) reference channel

photodiode. (Reprint from Levin 35 with permission from the Springer Nature.

Copyright 2005.)

Figure 5: The Nanodrop sample retention system. (a) one microliter of sample is loaded

directly onto the lower measurement surface. (b) The sample is engaged an upper

optical surface and retained by surface tension during the measurement. (Reprint from

ThermoFisher Scientific web page 97 )

Figure 6: Schematic (not to scale) of the GC-VUV instrument. (Reprint from Schug et

al. 98 with permission from the American Chemical Society. Copyright 2014.)
Table 1 Approximate wavelengths, energies and type of excitation for different spectral

regions. (Reprint from Tissue 9 with permission from the John Wiley and Sons,

Copyright 2012)

Spectral Wavelength Energy range Energy range Types of

region range (nm) (cm-1) (eV) excitation

Vacuum-UV 10-180 1x106-56,000 120-6.9 Electronic

UV 180-400 56,000-25,000 6.9-3.1 Electronic

Visible 400-750 25,000-13,300 3.1-1.6 Electronic

Electronic,
NIR 750-2,500 13,300-4,000 1.6-0.50 vibrational
overtones

Vibrations,
IR 2,500-25,000 4,000-400 0.50-0.050
phonons