Recent developments in derivative ultraviolet/visible absorption ...

Abstract
Review
Recent developments in derivative ultraviolet/visible
absorption spectrophotometry
C. Bosch Ojeda, F. Sanchez Rojas ∗
Department of Analytical Chemistry, Faculty of Sciences, University of Málaga, 29071 Málaga, Spain
Received 19 February 2004; received in revised form 18 May 2004; accepted 18 May 2004
Derivative spectrophotometry is an analytical technique of great utility for extracting both qualitative and quantitative information from
spectra composed of unresolved bands, and for eliminating the effect of baseline shifts and baseline tilts. It consists of calculating and plotting
one of the mathematical derivatives of a spectral curve. Thus, the information content of a spectrum is presented in a potentially more useful
form, offering a convenient solution to a number of analytical problems, such as resolution of multi-component systems, removal of sample
turbidity, matrix background and enhancement of spectral details. Derivative spectrophotometry is now a reasonably priced standard feature
of modern micro-computerized UV/Vis spectrophotometry.
The instrumental development and analytical applications of derivative UV/Vis regions absorption spectrophotometry produced in the last
10 years (since 1994) are reviewed.
© 2004 Elsevier B.V. All rights reserved.
Keywords: Derivative UV/Vis; Multi-component analysis; Partial least-square regression analysis; Multiple linear regression
1. Introduction
Derivative spectrophotometry, which consists in the differentiation
of a normal spectrum, offers a useful means for
improving the resolution of mixtures, because it enhances
the detectability of minor spectral features. Derivative spectrophotometry
is an analytical technique of great utility for
extracting both qualitative and quantitative information from
spectra composed of unresolved bands by using the first or
higher derivatives of absorbance with respect to wavelength.
It tends to emphasize subtle spectral features by presenting
them in a new and visually more accessible way, allowing
the resolution of multi-component elements, and reducing
the effect of spectral background interferences.
This technique offers an alternative approach to the enhancement
of sensitivity and specificity in mixture analysis.
It consists of calculating and plotting one of the mathematical
derivatives of a spectral curve. The derivative transformation
does not increase the information content of a
spectrum, but it permits discrimination against broad band
∗ Corresponding author. Tel.: +34-952131925; fax: +34-952132000.
E-mail address: fsanchezr@uma.es (F. Sanchez Rojas).
0003-2670/$ – see front matter © 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2004.05.036
interferences, arising from turbidity or non-specific matrix
absorption.
The derivative spectra are always more complex than the
zero-order spectrum. The first derivative is the rate of change
of absorbance against wavelength. It starts and finished at
zero, passes through zero at the same wavelength as λmax of
the absorbance band with first a positive and then a negative
band, with the maximum and minimum at the same wavelengths
as the inflection points in the absorbance band. This
bipolar function is characteristic of all the odd-order derivatives.
The most characteristic feature of the second-order
derivative is a negative band with the minimum at the same
wavelength as the maximum on the zero-order band. It also
shows two additional positive satellite bands on either side of
the main band. The fourth derivative shows a positive band.
The presence of a strong negative or positive band, with
the minimum or maximum at the same wavelength as λmax
of the absorbance band, is characteristic of the even-order
derivatives. Note that the number of bands observed is equal
to the derivative order plus one.
In two reviews published some years ago [1,2], weexposed
the different aspects of derivative ultraviolet/visible
spectrophotometry (DS): theoretical, instrumental devices

2 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
and analytical applications, the first until 1986 and the second
until 1993.
The purpose of this paper is to review the articles on
the anterior cited aspects published since 1994, in order to
complete the revision since the publication of our last review.
2. Theoretical and instrumental aspects
Derivative spectra can be obtained by optical, electronic
or mathematical methods. Optical and electronic techniques
were used on early UV/Vis spectrophotometers but have
largely superseded by mathematical techniques. The advantages
of the mathematical techniques are that derivative spectra
may easily be calculated and recalculated with different
parameters, and smoothing techniques may be used to improve
signal-to-noise ratio.
The main optical technique is wavelength modulation,
where the wavelength to the incident light is rapidly modulated
over a narrow wavelength range by an electrochemical
device, the first and second derivatives may be generated using
this technique. First derivative spectra may also be generated
by dual-wavelength spectrophotometer, the derivative
spectrum is generated by scanning with each monochromator
separated by a small constant wavelength difference.
First- and higher-order derivatives can be generated using
analogue resistance–capacitance devices. These generate
the derivative as a function of time while the spectrum
is scanned at constant speed. The electronic method suffers
from the disadvantage that the amplitude and wavelength
shifts of the derivatives vary with scan speed, slit width and
resistance–capacitance gain factor.
To use mathematical techniques, the spectrum is first digitalized
with a sampling interval of λ, the size of λ will
be dependent upon the natural bandwidth of the bands being
processed and of the instrumental bandwidth of the instrument
used to generate the data [3].
Instrumental requirements for derivative spectroscopy
are, in general, similar to those for conventional absorbance
spectroscopy, but wavelength reproducibility and
signal-to-noise ratio is of increased importance. The increased
resolution of derivative spectra puts greater demands
on the wavelength reproducibility of the spectrophotometer.
Small wavelength errors can result in much larger signal
errors in the derivative mode than in the absorbance mode.
The negative effect of derivatization on signal-to-noise also
places increased demands on low-noise characteristics of
the spectrophotometer. It is advantageous in this case if the
spectrophotometer can scan and average multiple spectra
to further improve the signal-to-noise ratio before derivatization.
For the derivatization process, it is important to be
able to control the degree of smoothing that is applied in
order to adapt to differing analytical problems. In the case
of the Savitzky-Golay method this means being able to vary
the order of the polynomial and the number of data points
used.
One kind of mathematical method, fractional calculus,
is useful for analyzing optical spectra. A new fractional
derivative spectroscopy is proposed [4]. In other paper, titled
“UV-visible absorption spectroscopy. Dual and second
derivative. Comparative study”, the techniques were compared
on the example of Almidon and Rhodamine B [5].
The possibility of reducing the noise in second derivative
without loss of spectral information is investigated. The classical
procedures for noise elimination curve smoothing or
increasing the differentiation step lead to partial loss of spectral
information. A simple method for noise reduction called
“step by step” filter (SBSF) is proposed [6,7]. A microcomputer
program, based on SBSF, was developed for calculation
of derivative curves directly from spectra recorded
as a function of wavelength. This program avoids the long
wavelength attenuation featured at conventional method for
derivative curves calculation, and in this extent could be
helpful for daily spectroscopy practice. The features of the
SBSF program include: easy treatment of data through a
windowed environment, calculating both conventional and
step by step filter derivatives, possibilities for selection of the
mathematical conditions for smoothing and differentiation
simultaneous plotting of the original curve and its derivative,
and a mouse pointer. Several examples from different
branches of the molecular spectroscopy are provided and
discussed in terms of advantages of SBSF [8].
A mathematical program in Turbo Pascal 6.00, which
is enable to generate spectra and to obtain first and second
derivatives is proposed. The second derivative of salicylic
acid showed a trough at 292 nm and a satellite peak at
316 nm. When large amounts of aspirin coexisted, the trough
disappeared, but the height of satellite peak was not altered
even at an aspirin concentration which was 2000 times more
than that of salicylic acid (corresponding to salicylic acid
content of 0.05%). The results were compared with those
obtained by using a spectrophotometer for recording the
second-order derivative spectrum of aspirin [9].
The complicated electronic absorption spectra of several
picramide autocomplexes were interpreted by the second
derivative method. A linear correlation was established between
the values of the intramolecular charge-transfer energy
and the ionization potential of a donor fragment of
molecules. The ionization potentials of several compounds
were determined [10].
The second derivative transformation of the absorption
spectrum of trimethylamine is extremely effective in enhancing
the information about the vibrational characteristics of
the excited state of the molecule, lying obscure in the raw
absorption data [11].
A derivative method combined with Fourier least-square
fitting was designed to process noised overlapped peaks. After
being fitted with least-square method, data were treated
with fractional derivative to amplify peaks. A formula was
used to rectify the shift of peak position in derivative spectrum.
Simulated and true UV spectral signal were processed
and the results were satisfied [12].

The new method “dynamic derivative spectroscopy”
(DDS) is presented for contamination detection and monitoring
in water. As a first application, the selective measurement
of aromatic solvents is investigated. The feasibility of
the new technique to measure such substances selectively
at competitive measurement speed is demonstrated. In conventional
transmission spectroscopy, the light transmitted
through the sample is attenuated in a specific manner by
the contained absorbers. In trace measurements, this attenuation
is very weak and overlaid by huge offsets due to
the emission spectrum of that radiation source. This leads
to problems due to the limited dynamic range of the electronics.
Additionally, in the UV region many absorption
features are broad, sometimes rather similar and overlapping.
This makes discrimination of different substances
difficult. The DDS technique makes use of a wavelength
modulation that generates optically an approximation of the
first and second derivatives with respect to the wavelength
of the transmission spectra. These derivatives are used for
evaluation in addition to the conventional transmission signal
in order to enhance spectral features for discrimination
of the compounds contained in the samples [13].
Several reviews have been published, in the last years,
on the principle and application of UV-visible spectrometry
[14–17].
2.1. Multi-component analysis
Multi-component analysis has become one of the most
appealing topics for analytical chemists in the last few years.
In this context, simultaneous determinations have proved
rather useful for resolving mixtures of analytes of interest
in such diverse fields as clinical chemistry, drug analysis,
pollution control, etc. The accuracy of the results obtained
in multi-component analyses by application of multivariate
calibration to the absorbance signals depends on the particular
method and the analytical signal used.
A method is described for the determination of the protein
mixtures using fourth derivative, the methodology was
demonstrated by quantifying mixtures of the three main
bovine caseins, the 244–296 nm spectra region was used for
the development of prediction models using the multivariate
method of partial least-square (PLS) regression analysis
[18].
A multivariate calibration method, PLS (types 1 and 2),
was applied to the simultaneous determination of naptalam
[N-(1-naphthyl)phthalamic acid] and its metabolites
N-(1-naphthyl)phthalimide and 1-naphthylamine in mixtures
by UV–vis absorption spectrophotometry. The absorption
and first derivative absorption spectra of mixtures were
used to perform the optimization of the calibration matrices
by the PLS method. Two different experimental designs for
the three-component mixtures are assayed and the results
are discussed. The proposed method with the derivative
spectra was applied to the determination of these analytes
in river water at the gl −1 level [19].
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 3
The performance of two multivariate calibration methods,
multiple linear regression (MULTI3) and PLS-2 to
absorbance and derivative spectrophotometric signals, for
the resolution of the ternary mixtures of acetylsalicylic
acid–caffeine–codeine and acetaminophen–caffeine–codeine
is compared and the proposed method was successfully
demonstrated for pharmaceutical tablets [20].
Least-square methods, classical least-squares (CLS),
Kalman filtering and PLS were compared for the analysis
of normal and derivative absorption spectra of some veterinary
sulfa drugs with two different calibration designs
and in two sets of wavelength range. Significant advantages
were found by the application of the derivative technique
coupled with the PLS method [21].
Four chemometric techniques, CLS, inverse least-squares
(ILS), principal component regression (PCR) and PLS were
applied to the absorption and first derivative spectrophotometric
determinations of amiloride and hydrochlorothiazide
in pharmaceutical preparation [22].
Brown et al. [23] were investigated the derivative preprocessing
as a method of drift noise reduction in multivariate
spectral data. This examination was carried out from the
perspective that baseline drift can be characterized as correlated
measurement errors, and that derivative filtering alleviates
some drift noise by reducing the covariance terms in
the error covariance matrices. While this approach is often
successful to some degree, derivative filters cannot be considered
optimal, since the error covariance matrix can rarely
be diagonalized by their operation. In addition, the use of
derivative filters modifies the composition of the chemical
signals in a fashion that is very difficult to predict a priori,
making the effects on figures of merit in multivariate calibration
largely unpredictable.
Derivative filters operate blindly in reducing drift noise
and, therefore, must be chosen on a trial-and-error basis,
but maximum likelihood PCA uses error covariance information
obtained from replicate measurements to achieve the
simultaneous drift correction and the maximum likelihood
projection of the spectral data into a principal component
space. It was shown that MLPCA is the optimal filter from a
drift reduction perspective, since MLPCA uses error covariance
information to diagonalize the error covariance matrix
and thus eliminate drift noise. The regression counterpart to
MLPCA, MLPCR, is consequently an optimal calibration
method to use when drift noise plagues the acquired data.
Baseline drift poses a significant threat to the precision
and accuracy of many calibration methods. Derivative preprocessing
has been widely employed to combat this problem
in the past, but since it is suboptimal in terms of drift
correction, its application requires time-consuming searches
for the best filter characteristics for a given application. Unfortunately,
the spectral interpretability also suffers upon differentiation.
In this study, MLPCR was consistently found
to perform as well as or better than derivative PCR when
reasonable estimates of the error covariance structure were
available. It is therefore recommended that, provided that

4 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
error covariance information obtainable, MLPCR is used as
a calibration method for data corrupted by baseline drift.
2.2. Combination of DS with flow injection (FIA), liquid
chromatography and kinetic analysis
In liquid chromatography with photodiode array detection,
a number of approaches have been developed in
chemometrics to determine an analyte of interest in the
presence of one or more unknown compounds; so far,
these techniques have required time-consuming data analysis
programs, and are not suitable for routine analysis.
A “derivative spectrum chromatogram” is proposed as a
new approach for resolving overlapped peaks in real time.
This procedure is based on the following assumptions: (i)
an absorption spectrum of an analyte with rounded humps
changes to steep peaks by high-order derivatization and (ii)
the principle of superposition is valid assessed with real data
for the separation of pesticides by liquid chromatography
[24]. Other paper reports the possibilities of HPLC on-line
with photodiode array detection for identification of bovine
milk proteins, using second and fourth derivatives [25].
Application of SDS-polyacrylamide gel electrophoresis
and reversed-phase high-performance liquid chromatography
on-line with the second and fourth derivatives UV
spectroscopy in identification of -casein and its peptide
fractions was reported [26].
A flow injection procedure was developed for the simultaneous
determination of two UV filters (oxybenzone and
2-ethylhexyl-4-methoxycinnamate) in sunscreen formulations,
based on the isodifferential approach, using second
derivative [27].
Simultaneous dissolution profiles of two drugs (sulfametoxazole
and trimethoprim) in pharmaceutical formulations
by an FIA manifold are proposed. The official procedure is
adapted to the continuous flow methodology; the dissolution
vessel is connected to an FIA manifold, the simultaneous
determination of both profiles is based on the first derivative
and the zero-crossing mathematical procedure [28].
The possibility has been investigated by applying derivative
analysis to a classical enzymatic-spectrophotometric
method for lecithin determination for the purpose of developing
an analytical direct method that does not require long
pretreatment of the test sample even in the case of turbid
samples [29].
A review shows the potentiality (but also the limits)
of the UV derivative spectroscopy specially adapted to
high-pressure expositions [30].
3. Applications
3.1. Inorganic analysis
DS involves calculating and plotting one of the mathematical
derivatives of the spectral curve offers and alternative
approach to metal analysis, while at the same time showing
good sensitivity and specifity. In the derivative spectrum, the
ability to detect and to measure minor spectral features is
considerably enhanced.
Derivative UV absorption spectroscopy is a powerful
spectroscopic technique for multi-component gas analysis,
particularly in combustion and process controlling application
[31].
The analysis of anions has hardly been investigated by
means of derivative spectroscopy; a third derivative procedure
is proposed for the determination of traces of phosphate,
based on the formation of a ternary ion-association
complex by reaction of phosphate with rhodamine 6G in the
presence of molybdate [32]; second derivative was applied
to simultaneous determination of nitrate and nitrite ions in
bath solutions for alkaline black-oxidation of steel [33]. A
direct and simultaneous UV second derivative was proposed
for the determination of nitrite and nitrate in preparations
of peroxynitrite [34], the determination of nitrate nitrogen
in groundwater by first-order derivative has been described
[35] and the evaluation of second derivative for nitrate and
total nitrogen analysis of wastewater samples is also reported
[36]. The second derivative UV absorbance method used by
Crumpton et al. (1992) for analyzing NO3 − in fresh water
was explored as a simple, quick and reliable method for
measuring NO3 − depletion in the MPN culture medium. In
a complex matrix such as a culture medium, the individual
components are often indistinct in UV absorbance spectra
because of the large widths of component bands relative to
the separations between adjacent bands. In some cases, the
remedy is to calculate and plot first, second and possibly
higher derivatives of the absorbance spectrum with respect to
wavelength. A second derivative method proved to be a fast
and reliable means for measuring nitrate depletion in MPN
media used for enumerating autotrophic and heterotrophic
nitrate-reducing bacteria, without interferences from Cl − or
the organic components in the latter medium [37].
The simultaneous determination of phosphate and silicate
in material samples utilizing a simple, rapid and inexpensive
method is difficult, due to the mutual interference between
the two anions. A first derivative spectrophotometric
method utilizing the zero-crossing measurement technique
was developed for the simultaneous determination of
phosphate and silicate in synthetic detergents and water,
based on the formation of phospho- and silicomolybdenum
blue complexes in the presence of metol or ascorbic acid
[38].
Also, an approach for quantitative spectrophotometric
analysis of binary mixtures based on the derivative
spectra has been developed. The proposed method is
used for investigation of the complex formation of some
aza-15-crown-5-containing chromoionophores with Sr 2+
where the stability constant is low and the absorption spectra
of the complex cannot be obtained directly [39].
The different determinations achieved are described in
Table 1.

C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 5
Table 1
Determination of metal ions
Elements Reagents Remarks Reference
Ag 2-Nitroso-1-naphthol-4-sulfonic acid
(nitroso-S) and
tetradecyldimethylbenzylammonium
chloride (TDBA)
Fourth derivative, preconcentration on
microcrystalline naphthalene or column method,
0.2–30 g ml −1 , in various complex samples
Ag, Au Rhodamine derivative First and second derivatives using zero-crossing
technique, micellar medium, in silicate rocks
[41]
Al Hydroxynaphthol blue First derivative, 11.8–320.0 ng ml−1 [42]
Al Oxine-5-sulfonic acid In stomach medicine [43]
Al Chromazurine S Second derivative at 543 nm, sensitivity
enhancement of oil/water microemulsion
[44]
Al CAS Sensitizing effect of nonionic microemulsion on
the derivative
[45]
Al Aluminon pH 3.62 Clark-Lubs buffer, λ 362 nm, Al is
determined in water
[46]
Al, Fe Hematoxilin First and second derivatives zero-crossing method,
in the presence of surfactant, in glasses, phosphate
rocks, cement and magnesite alloy
[47]
Am – Second derivative at 503.5 nm, in a nitric acid
medium, 1–11 g ml−1 [48]
Au, Pd 3-Hydroxy-2-methyl-1-phenyl-4-pyridone Third derivative, 1–13.0 and 0.28–8.0 g ml−1 ,
respectively
[49]
Bi 2-(5-Bromo-2-pyridylazo)-5-
Third derivative, separation and preconcentration
[50]
diethylaminophenol (5-Br-PADAP)
with ammonium tetraphenylborate on
microcrystalline naphthalene or by column
method, in alloys and synthetic samples
Bi, Sb Iodide First derivative, in synthetic samples [51]
Ca, Mg CAS and CTMAB Ratio spectrum and derivative, in synthetic and
real water samples
[52]
Cd, Co, Cu Meso-tetrakis[4-
Third derivative at 412, 434 and 438 nm for Cu,
[53]
trimethylammonium)phenyl]porphine
Co and Cd, respectively, in synthetic mixtures
Co 5-Br-PADAP In several nickel matrices [54]
Co 1-Nitroso-2-naphthol Neutral micellar medium, selective determination
of Co in the presence of Cu(II) or Fe(III), Co is
determined in drugs and alloys
[55]
Co Disodium
Third derivative, on microcrystalline naphthalene,
1-nitroso-2-naphthol-3,6-disulfonate and
tetradecyldimethylbenzylammonium
chloride
0.1–11 g ml−1 [56]
, in alloys and biological samples
Co, Cu Pyridoxal-4-phenylthiosemicarbazone First derivative, use of a flow-through sensor, in
several steel samples
[57]
Co, Fe Meso-tetrakis(4-sulfophenyl)porphyrin First derivative, 0–0.24 and 0–0.18 mg ml−1 ,
respectively, in tobacco
[58]
Co, Ni 4-(2-Pyridylazo)resorcinol Second derivative, 0.25–1.50 and 0.20–1.25 ppm,
respectively
[59]
Co, Ni Dithiazone First derivative at 620 and 740 nm, 5.0 ×
10−6−1.0 × 10−4 and 2.0 × 10−5−2.0 × 10−4 M,
respectively, applied to a Ni/Cr-based dental alloy
[60]
Co, Mn, Zn 5-Br-PADAP Linear regression multiwavelength derivative, in
rice and wheat flour
[61]
Cr Eriochrome Cyanine-R Fourth derivative at 545 nm, 20–80 ng ml−1 , in steel [62]
Cr, Zr 2-(2-Pyridylmethyleneamino)phenol First derivative, in Zr/Cr bronzes [63]
Cu 3-(4-Phenyl-2-pyridinyl)-5-phenyl-1,2,4-
First derivative, solvent extraction in
triazine and picrate(2,4,6-trinitrophenol)
1,2-dichloroethane, 7.5-350 ng ml−1 [64]
, in several
kinds of water
Cu 1-(2-Pyridylazo)-2-naphthol (PAN) In non-ionic micellar medium, Cu content in
beverages, biological and alloy samples
[65]
Cu 2-Nitroso-1-naphthol-4-sulfonic acid
Second derivative, with
[66]
(nitroso-S)
tetradecyldimethylbenzylammonium chloride on
microcrystalline naphthalene, in standard alloys
and biological samples
Cu, Fe 5-Phenyl-3-(4-phenyl-2-pyridinyl)-1,2,4-
First derivative using the zero-crossing approach,
triazine (PPT)
solvent-extraction in dichloroethane, 50–2500 and
2–120 ng ml−1 [67]
, respectively, in well water
[40]

6 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 1 (Continued )
Elements Reagents Remarks Reference
Cu, Fe Picrate Second derivative zero-crossing approach for Cu
and graphic method for Fe, extraction in DCE
with PPT for Fe and bathocuproine for Cu, in
river and tap water
Cu, Fe 2-Hydroxy-1naphthaldehydebenzoylhydrazone
(OHNABH)
First derivative, 0.16–4.80 g ml−1 at 443 nm and
0.14–4.20 g ml−1 at 540 nm, respectively, in plant
samples
Cu, Hg, Pb Dithizone First derivative, extraction in chloroform, in
marine sediments samples
[70]
Cu, Ni Diacetylmonoxime benzoylhydrazone First derivative, in alloys [71]
Cu, Ni Diacetylmonoxime isonicotinoyl hydrazone Second derivative use of peak-to-base line
measurement techniques
[72]
Cu, V OHNABH First derivative at 443 and 465 nm, 0–4.5 and
0–7.5: g ml−1 , Cu in food materials, V in alloy
steels, and both V and Cu in Cr–V steel
[73]
Cu, Zn Meso-tetrakis(4-methoxyphenyl)porphine In human hair and serum samples [74]
Er, Nd 2-(Diphenylacetyl) indan-1,3-dione and
Third derivative, in mixed rare-earths and
[75]
dodecyl benzenesulfonic acid sodium-salt
lanthanide-based samples
Er, Ho, Nd Norfloxacin Second derivative, 5.0 × 10−5 to 2.5 × 10−4 Mof
Nd, Ho and Er, in mixtures of rare earth elements
[76]
Er, Ho, Nd 1-Ethyl-6,8-difluoro-7-(3-methyl-1-
Second derivative, 5.0 × 10
piperazinyl)-4-oxo-1,4-dihydro-3-quinoline
carboxylic acid
−5 to 2.5 × 10−4 M
[77]
of Nd, Ho and Er, determination of Nd, Ho and
Er in rare earth mixtures
Fe PAN Third derivative, preconcentration on a membrane
filter in the presence of capriquat
[78]
Fe Sulfosalicyl acid Third derivative, in some chemical reagents [79]
Fe Sulfosalicyl acid Sensitizing effect of nonionic microemulsion [80]
Fe Phenanthroline Second derivative, in foods [81]
Fe 2,4,6-Tripyridyl-1,3,5-triazine Second derivative at 660 nm, on solid phase, in
water
[82]
Fe 1,2-Dihydroxybenzene-3,5-disulfonic acid
Eighth derivative, separation and preconcentration
[83]
(Tiron)
on an adsorbent
tetradecyldimethylbenzylammonium chloride on
naphthalene as a slurry or packed in a column, in
standard alloys and biological samples
Fe, Mo Morin First derivative using zero-crossing method, in the
presence of a cationic surfactant, 0.9–1.5 and
0.3–4.2 mg ml−1 , respectively, in Co/Cr and Ni/Cr
alloys
[84]
Fe, Mo, Fe, V Cyclic hydroxamic acid Extraction with MIBK, comparison of derivative
and PLS(1 and 2) methods
[85]
Fe, Ni Xylenol orange and CTMAB First derivative, in Fe/Ni alloy plating layer [86]
Fe, Ni 4-(2’-Benzothiazolylazo)salicylic acid
First derivative, 2.1–8.4 and 0.59–7.08 g ml
(BTAS)
−1 ,
[87]
respectively, in steel alloys and aluminum alloys
Fe, Ru 4,7-Diphenyl-1,10-phenanthroline Zero-crossing approach, 9.6–450 and
16.3–280 g l−1 , respectively, determination of Fe
and Ru in synthetic mixtures
[88]
Fe, V 2-Hydroxy-1-naphthaldehyde
First derivative at 540 and 465 nm, 0.14–4.20 and
benzoylhydrazone
0.12–2.50 g ml−1 [89]
, respectively, in natural samples,
food and biological materials
Ga, In PAN First derivative using the isodifferential points at
550 and 542 nm, 0.28–3.63 and 0.46–9.20 g ml−1 ,
respectively
[90]
Ga, In 5-Br-PADAPl In cationic micellar medium, 0.023–0.700 and
0.076–1.52 mg ml−1 , respectively, in synthetic
binary mixtures, standard reference materials and
environmental samples
[91]
Ge Phenylfluorone-cetylpyridinium chloride Second derivative peak area, 0–15 g ml−1 ,in
Chinese herbs
[93]
Hg, Pd 5-(3,4-Methoxyhydroxyphenylmethylene)-
First derivative zero-crossing technique, 0.4–1.4
2-thioxo-1,3-thiazolidine and
cetyltrimethylammonium bromide
and 0.08–1.8 mg ml−1 [94]
, respectively
Ge 2,6,7-Trihydroxysalicylflourone Fourth derivative by compressing X-factor,
0.0008–0.0040 g ml−1 , in geochemical samples
[92]
[68]
[69]

Table 1 (Continued )
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 7
Elements Reagents Remarks Reference
Ho 2-(Diphenylacetyl)indan-1,3-dione and octylphenyl
poly(ethyleneglycol)ether
Second derivative, in rare earth mixtures [95]
Ir 1-(2-Pyridylazo)-2-naphthol First derivative, preconcentration on
microcrystalline naphthalene, in synthetic samples
corresponding to various standard alloys and
environmental samples
[96]
Ir 2-(2-Thiazolyazo)-5-dimethylaminophenol (TAM) First derivative, Ir–TAM complex is retained on the
tetraphenylborate-naphthalene adsorbent packed in
a column, in synthetic samples and standard alloys
[97]
Ir, Rh 2-(5-Bromo-2-pyridylazo)-5-diethylaminophenol
Zero-crossing approach, preconcentration onto
[98]
and tetraphenylborate
microcrystalline naphthalene, Rh and Ir are
determined in synthetic samples
Mn 5-Br-PADAP Second derivative at 554 nm, in aluminum and its
alloy
[99]
Mn 5-Br-PADAP-ammonium tetraphenylborate (TPB) Third derivative, separation and preconcentration
with microcrystalline naphthalene or by a column
method
[100]
Mo Hydrogen peroxide At 376 nm, in U–Mo alloy [101]
Mo, Zr Alizarin Red S First derivative at the zero-crossing wavelengths at
446.0 and 490.5 nm, 0.5–13.0 and 0.5–20.0 g ml−1 ,
respectively
[102]
Nb, Ti Hydrogen peroxide and 5-Br-PADAP Fifth derivative, in standard steel and Buffalo
River Sediment
[103]
Nd Bromopyrogallol red Fourth derivative, in the presence of TX-100, in
mixed rare earths
[104]
Nd Methyl Thymol Blue and cetylpyridinium chloride Fourth derivative, in mixed rare earths,
0–3.5 g ml−1 [105]
Nd Enoxacin and cetyltrimethylammonium bromide
Second derivative, 1.2 × 10
(CTMAB)
−5 to 2.7 × 10−4 M,
[106]
in mixed rare earth oxides
Ni Hydroxynaphthol blue First derivative, 21–800 ng ml−1 , in standard
brasses
[107]
Ni 5-Br-PADAP First derivative using zero-crossing method, in the
presence of Triton X-100, in steels standards
[108]
Ni 2-(5-Bromo-2-pyridylazo)-5-phenol In aluminum and aluminum alloys [109]
Ni, Pd 2-(2-Thiazolyazo)-5-dimethylaminobenzoic acid First derivative, 0.07–1.60 and 0.12–1.75 g ml−1 ,
respectively
[110]
Ni, Zn 2-(2-Pyridylmethyleneamino)phenol First derivative using zero-crossing technique,
0.3–3.0 and 1.0–6.0 g ml−1 , respectively, in a real
bronze sample
[111]
Os Thiourea Second derivative at 605 nm, in standard plasma
(ICP and MIP) solutions of noble metals
[112]
Os, Ru – Third derivative, in hydrochloric acid [113]
Pb Meso-tetra-(3,5-dibromo-4-hydroxyphenyl)porphyrin
Second derivative, in clinical samples [114]
Pd Pyridopyridazine dithione (PPD) Fourth derivative, in the presence of non-ionic
surfactant, in activated charcoal
[115]
Pd 1-(2-Pyridylazo)-2-naphthol support on naphthalene First derivative, column preconcentration, Pd is
determined in alloys and biological materials
[116]
Pd Disodium-1-nitroso-2-naphthol-3,6-disulfonate and Third derivative, on microcrystalline naphthalene,
tetradecyldimethylbenzylammonium chloride
0.4–37 g ml−1 [117]
Pd, Rh 5-(3,4-Methoxyhydroxybenzylidene)rhodamine Fourth derivative, cationic surfactants, in synthetic
mixtures
[118]
Pd, Rh 5-(2,4-Dihydroxybenzylidene)rhodamine First derivative, Pd and Rh in silicate rocks and
Pd in a Pd-activated charcoal
[119]
Pd, Pt 3-(2’-Thiazolyazo)-2,6-diaminopyridine-(2,6-
Second derivative, the zero-crossing and the
TADAP)
graphic methods were used for Pt and Pd,
respectively, 8.9 × 10−7 to 3.1 × 10−5 M for Pt
and 4.6 × 10−7 to 6.8 × 10−5 [120]
M for Pd
Pd, Pt Iodide First and second derivative spectra for Pd and Pt,
respectively, in a silver alloy
[121]

8 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 1 (Continued )
Elements Reagents Remarks Reference
Pd, Rh, Pt Tin(II) chloride In aqueous solutions or after extraction into
1,2-dichloroethane in the form of the ion-associates
Rh(Pd, Pt)–SnCl3–diantipirylmethane (DAM)
[122]
Pd, Zn PAR As a tool for determination of stability constants
of Zn–PAR and Pd–PAR complexes
[123]
Pr Norfloxacin Second derivative, determination of Pr in mixed
rare earths
[124]
Pr Lomefloxacin Second derivative, 3.5–65 g ml−1 , in mixed rare
earths
[125]
Pr Ciprofloxacin Third derivative, in mixtures of rare earths [126]
Pt Iodide Fourth derivative, in Pt–Ru/C catalyst [127]
Pt, Ru SnCl3-ligands(tin(II) chloride in HCl) First derivative at 377 nm (zero-crossing point of
Ru) and second derivative at 495 nm (zero
crossing point of Pt), Pt and Ru are determined in
Pt–Ru/C catalyst
[128]
Pu, U – First derivative of the ratios of their direct
absorption spectra at 473.8 and 411.2 nm, 0.5–2 and
10–25 mg g−1 , respectively, in 1 M HNO3 medium
[129]
Pu, U – First derivative of the ratios of spectra at 473.8
and 411.2 nm, respectively
[130]
Pu, U Arsenazo III First derivative at 632 nm for U and at 606.5 nm
for Pu, U and Pu are measured in the same
aliquot in fairly high burn-up fuels
[131]
Rh PAN First derivative, preconcentration of its complex
on microcrystalline naphthalene, determination of
Rh in synthetic samples
[132]
Rh 5-Br-PADAP and tetraphenylborate Third derivative, preconcentration on
microcrystalline naphthalene, determination of Rh
in synthetic samples and alloys
[133]
Rh 2-(2-Thiazolyazo)-5-dimethylaminobenzoic
Third derivative, Triton X-100 as surfactant,
acid (TAMB)
0.03–0.4 g ml−1 [134]
Rh, Ru Octadecyldithiocarbamate First derivative, 1.0–10.0 and 0.5–6.0 g ml−1 ,
respectively
[135]
Sc 1-(2-Thiazolyazo)-2-naphthol Second derivative, column preconcentration onto
tetraphenylborate–naphthalene adsorbent,
0.08–2.8 g ml−1 , in standard biological and
synthetic samples
[136]
Sm Arsenazo III Second derivative at 655 and 677 nm, in Ni/Fe
alloy electroplating
[137]
Th 2,4-Dihydroxybenzaldehyde isonocotinoyl
hydrazone (2,4-DHBINH)
First derivative, 0.30–7.00 g ml−1 [138]
Ti 2,4-Dihydroxybenzaldehyde isonicotinoyl
hydrazone
First derivative, in several alloy and steel samples [139]
U – Second derivative at 408.2 nm, in magnesium
diuranate (Yellow Cake)
[140]
U 1,4-Dihydroxy-9,10-anthracenedione Second derivative, in a cationic micellar medium [141]
U, V PAR First derivative at 563 and 600 nm, 0.4–4.0 and
4.0–16 g ml−1 , respectively, in the presence of
cetylpyridinium chloride and EDTA, in synthetic
matrices, river and saline water samples
[142]
V 5-Br-PADAP Second derivative, adsorptive extraction onto
microcrystalline benzophenone, in alloys and
environmental samples
[143]
V 5-Br-PADAP and ammonium
Third derivative, preconcentration with
[144]
tetraphenylborate
microcrystalline naphthalene or by column method
W Thiocyanate Second derivative, determination of W in
niobate-tantalates, tin slag samples, ores,
concentrates and vanadium and molybdenum
bearing geological materials
[145]
Zn Meso-tetrakis(4-aminophenyl)porphyrin Second derivative, in water [146]
Zn Meso-tetra-(3,5-dibromo-4hydroxyphenyl)porphyrin
Second derivative, in serum [147]
Zr 2,4-DHBINH First derivative [148]

3.2. Organic compounds
Spectrophotometric methods of analysis have experienced
a high evolution in the last 25 years and widely used as a tool
for quantitative analysis, characterization and quality control
in agricultural, pharmaceutical and biomedical fields.
Derivative UV spectroscopy which is based on mathematical
transformation of spectral curve into derivative spectra
eliminates the influence of background or matrix and usually
provides much better fingerprints than the traditional
absorbency spectra. The focal point of applications in organics
lies in aromatic amino acids, proteins such as enzymes,
pharmaceuticals in various forms, and natural and synthetic
pigments and dyestuffs.
Binary and ternary mixtures of organic compounds have
been widely analyzed using DS. Although pharmaceutical
formulations are the mixtures most frequently assayed, and
are especially considered in the next section, other diverse
mixtures as aromatic hydrocarbons, antioxidants, phenols,
herbicides, etc., have been determined with good results.
For example, spectra of diverse derivatives of phenol overlap
strongly in the whole analytical region and the direct
application of spectrophotometry is impossible. DS allows
separating these spectra and determining selected groups of
phenols.
A second derivative UV-Vis absorption method to follow
kinetics of organic reactions, when two bands are strongly
overlapped in the fundamental spectra, is described [149].
Table 2 shows the diverse procedures described since
1994.
3.3. Pharmaceutical analysis
Compared with conventional spectrophotometric determinations,
derivative spectrophometry has proved to be a great
value in eliminating the interference from excipients and coformulated
drugs. The derivative spectrophotometry represents
an elegant approach to the problem of resolving spectral
overlap in pharmaceutical analysis. DS permits the simple,
rapid, sensitive and direct determination of mixtures of
drugs having closely overlapping spectra.
In pharmaceutical application, derivative spectrophotometry
has led to significant developments in the analysis of
drugs in the presence of their degradation products or in
multi-component mixtures.
The application of DS in pharmaceutical analysis has very
important.
The reaction of benzylpenicillin (I) with CrCl3 was studied
by IR, DS and elemental analysis of the products. Elemental
analysis at I/Cr 3+ ratios of 2:1, 3:1 and 3:2, indicated
3:2, or a multiple thereof, as the most likely interaction ratio,
spectroscopy indicated that the product is likely a dimer,
in which Cr 3+ has a pseudo-octahedral geometry [173].
Second derivative was used for determination of partition
coefficients of chlorpromazine and promazine between
lecithin bilayer vesicles and water [174], phenothiazine
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 9
derivatives (chlorpromazine, triflupromazine, promazine,
promethazine, trifluoperazine and prochlorperazine) between
human erythrocyte ghost membranes and water
[175], diazepam and flurazepam between phosphatidylcholine
bilayer vesicles and water [176], two nonsteroidal
anti-inflammatory drugs (indomethacin and acemetacin)
between Egg yolk phosphatidylcholine multilamellar
vesicles and water [177], chlordiazepoxide (benzodiazepine),
isoniazid and rifampicin (tuberculostatic drugs)
and diucaine (local anesthetic) between lipid bilayers of
dimyristoyl-l-alpha-phosphatidylglycerol unilamellar liposomes
and water [178] and phenothiazine drugs (trifluoperazine,
triflupromazine, chlorpromazine and promazine)
between phosphatidylcholine small unilamellar vesicles and
water [179].
Second derivative spectrophotometric study on the interactions
of chlorpromazine and triflupromazine with bovine
serum albumin was proposed [180].
First derivative was used for the determination of the degree
of deacetylation of chitosan [181].
Kinetic study on the degradation of prazepam in acidic
aqueous solutions by high-performance liquid chromatography
and fourth derivative UV spectrophotometry was proposed
and evaluated [182], and the study of the photochemical
degradation of nisoldipine was performed under daylight
exposure by means of UV derivative [183].
The isomer distribution of camptothecin was studied by
first derivative ratio spectrum of UV–vis absorption and fluorescence
spectrometry in different pH aqueous buffer solutions.
It was found that camptothecin exists mainly in the
opened-ring form in physiological pH [184].
A first derivative method was developed for the determination
of nifedipine in oil/water/oil multiple microemulsions
during stability studies [185] and the stability study of
a naphthoquinone in cyclodextrin complexes by a validated
second derivative method was developed and evaluated for
the kinetic investigations [186].
Because of a rapid development of microcomputer technology,
DS became a practical analytical method in the
general laboratory. The great interest for this technique is
due to the increased resolution of the spectral bands, elimination
of interferences originating from sample turbidity
and matrix background and the enhancement of spectral
details. These characteristics led to wide DS application
in biochemical, pharmaceutical, food and environmental
analyses. However, less attention has been paid to the application
of this method in the studies of chemical equilibria.
Equilibrium transformations, especially acid–base ones,
sometimes involve minor changes in their electronic spectra
which makes it impossible to employ classical spectrophotometry
for determination of acidity constants. DS seems
to be suitable for solving this problem. A general derivative
spectrophotometric method for determination of acidity
constants is developed. The method appears suitable in
cases when classical spectrophotometry cannot be employed
due to little differences in the absorption spectra of conju-

10 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 2
Determination of organic compounds
Compounds Remarks Reference
Alkylphenol polyoxyethylene ether Second derivative, 25–700 g ml−1 , in DTY finishing oil [150]
Aromatic components: alkylbenzenes,
Second derivative, the role of derivative spectroscopy in the analysis of aromatics [151]
indans, tetralins and alkylnaphthalenes present in straight-run Kerosenes/ATF or reformed kerosenes is assessed
Aromatic hydrocarbons Second derivative, in gas oils [152]
Aromatic hydrocarbons BTEX in water [153]
Aromatic hydrocarbons First and second derivative of transmission spectra, combined with chemometric
algorithms like PCR or PLS, in water
[154,155]
Natural hydrocarbon Calibration with fourth derivative, 4.4–230.4 g ml−1 , in CHCl3 [156]
Barium benzene sulfonate Second derivative, in the presence of trace benzene in aqueous media [157]
Fungicide ferbam
Fourth derivative using 2,2
[iron(III)dimethyldithiocarbamate]
′ -bipyridyl complex in Triton X-100, 0.5–20 g ml−1 ,in [158]
a commercial sample and in mixtures with various dithiocarbamates (ziram,
zineb, maneb, etc.) And from wheat grains
Polynuclear aromatic hydrocarbons (PNA) Second derivative, several typical PNA, in diesel fuels, which are known
precursors of carcinogenic PNA in emissions
[159]
2-Mercaptobenzimidazole Second derivative at 304 nm and third derivative at 308 nm, in synthetic mixtures
of polymer additives and in rubber samples
[160]
2-Mercaptobenzimidazole Second derivative, in ternary synthetic mixtures of rubber additives and in rubber
samples
[161]
Novolac layers Thin solid layers of halophenol and cresol novolacs were investigated by UV
derivative
[162]
Parathion and p-nitrophenol First derivative at 253.0 and 273.1 nm, in vegetable tissues [163]
Permethrin Second derivative at 279 nm, in shampoo [164]
Phenols In water with back extraction—fourth derivative, 0–12 g ml−1 [165]
Methyl- and chlorophenols After solid-phase extraction preconcentration [166]
Phenols and herbicides Methyl- and chlorophenols (3-methylphenol, 2,3- and 3,4-dimethylphenol, 2,5-,
2,6- and 3,4-dichlorophenol and 2,4,5-trichlorophenol) and triazine, uracil and
urea herbicides (simazine, propazine, hexazinone, bromacil and metoxuron) were
examined
[167]
Phenyl-beta-naphthylamine Second and third derivatives, in rubber samples [168]
-Diketones Normal or derivative, zero-crossing point method, with 2,3-diaminonaphthalene
as reagent of derivatization
[169]
Rubber antioxidant
Third derivative for the elimination of the mutual interferences of some additives [170]
(phenyl-beta-naphthylamine, PBN)
in the PBN estimation
Triazinyl-2-pyrazolines The effects of the substituents of the s-triazine cycle upon UV absorption [171]
Undetonated explosives Characterization of some common undetonated explosives [172]
gated acid–base pairs. Based on theoretical considerations,
seven variants of the method have been established and
their validity was checked, determining acidity constants of
lorazepam and flurazepam as model compounds [187].
Applications of derivative UV spectroscopy to the analysis
of a representative set of drugs (baclofen, cimetidine,
cycloserine, diethyltoluamide, clobetasol propionate, clobetasone
butyrate and beclomethasone dipropionate) are
described, the derivative technique presents significant advantages
over the conventional absorption method [188].
Specific procedures are summarized in Table 3.
3.4. Analysis of biological compounds
A dual-wavelength differential first derivative method was
developed for identification and determination of carbon
monoxide generated from microsomal metabolism of xenobiotics
in vitro. Four trihaloanilines and one trihalobenzene
were tested using this method, the results showed that only
2,4,5-trifluoroaniline could be converted to CO by the incubation
with rat hepatic microsomes, NADPH and oxygen.
The ability of phenobarbital or dexamethasone to induce rat
hepatic microsomes to catalyze CO formation was 3–8 times
higher than that of the control [382]. Procedures described
since 1994 are shown in Table 4.
3.5. Food analysis
Quality control of extra virgin olive oils: UV spectrophotometric
analysis with first derivative absorbance spectra between
230 and 280 nm of extra virgin olive oils was used to
graphically present the oil residual shelf-life and conservation
status [419] and alumina cleanup: oxidized and refined
components [420].
The effect of dietary oregano oil and alpha-tocopheryl
acetate supplementation on iron-induced lipid oxidation
of turkey breast, thigh, liver and heart tissues and on the
inhibition of lipid oxidation in long-term frozen stored
chicken meat was examined by third derivative spectrophotometry
[421,422]. Procedures involving the application
of UV–vis derivative to food analysis are summarized in
Table 5.

Table 3
Analysis of compounds in pharmaceutical formulations
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 11
Compounds Remarks Reference
Acebutolol HCl First derivative at 266.6 nm, in the presence of its acid-induced
degradation product, to investigate the kinetics of the acid
degradation process at different temperatures
[189]
Acetylsalicylic acid Second derivative at 297 nm, 100–450 mg ml−1 , in effervescent
tablets
[190]
Acyclovir diloxanide furoate Second and third derivatives, acyclovir in the presence of
guanine (its main impurity) and diloxanide furoate in the
presence of diloxanide (its degradation product)
[191]
Alkaloids First derivative, in preparations of aconitum carmichaelii and
aconitum kusnezoffii
[192]
Amphotericin B Third derivative, in serum and urine [193]
Anafranil Difference and difference first and second derivatives [194]
Angiotensin-converting enzyme (ACE) inhibitors Ramipril (third), benazepril (second), enalapril maleate (second),
lisinopril (first and second) and quinapril (first), in
pharmaceutical dosage forms
[195]
Anti-inflammatory drugs First derivative [196]
Atorvastatin First derivative at 217.8 nm, 4.2–69.0 g ml−1 [197]
Baicalin Second derivative at 296 nm, in the Langin oral liquid [198]
Benzalkonium bromide Second derivative in disinfection solutions, the peak to peak
amplitude at 265 and 267 nm was taken for quantitative
determination
[199]
Benzenoid UV-absorbing drugs Second, third and fourth derivatives, identification and
differentiation between benzenoid UV-absorbing drugs
[200]
Benzophenones Using the isodifferential approach [201]
Bifonazole Second derivative, in the presence of methyl and
propyl-hydroxybenzoate
[202]
Buclizine hydrochloride In bulk and tablets form [203]
Butamyrate citrate First derivative at 253.6 nm, in cough syrups [204]
Carbinoxamine maleate Second derivative at 269 nm in the presence of pseudoephedrine
HCl in rhinostop oral drops
[205]
Cefotaxime (triethylammonium salt) In a reaction mixture in the presence of the related compounds
from synthesis, 0.005–0.080 mg ml−1 at 276.8 nm
[206]
Cephalexin First derivative at 254 nm, in oral suspensions containing
potassium sorbate, 5–100 mg ml−1 [207]
Cephalosporines: cefadroxil (I), cephradine (II)
and cefaclor (III)
At 345 nm for I and II and at 334 nm for III using zero-order
absorption curve, at 334 nm using first derivative spectrum for
III, in the presence of degradation products
Cetirizine dihydrochloride First and second derivative amplitudes at 239 (peak) and
243–233 nm (peak-to-trough), respectively, in bulk and tablet
form
[209]
Chlorpromazine hydrochloride Second derivative, use of internal standard method
(1,10-phenanthroline), Savitzky-Golay algorithm was used for
separation signals
[210]
Cinchocaine hydrochloride First derivative, in the presence of its acid-induced degradation
product
[211]
Ciprofloxacin Direct estimation in liposomes [212]
Cisapride First derivative at 264, 300 nm and second derivative at 276,
290 and 276–290 nm
[213]
Clobutinol First and second derivatives at 277 and 275 nm [214]
Clozapine First derivative, 5–50 g ml−1 [215]
Cyclofenil Second derivative, in the presence of its acid-induced
degradation product
[216]
Diclofenac sodium (degradation products) Second derivative at 260 and 265 nm for oxindol, in gel-ointment [217]
Droperidol First derivative at 255.2 nm, in the presence of parabens [218]
Fleroxacin First, second, third and fourth derivatives using peak–zero and
peak–peak methods
[219]
Fluconazole Second derivative at 274 nm [220]
Fluconazole First derivative at 271.6 nm, 126–462 g ml−1 , in syrups [221]
Fluoxetine First derivative, 5–30 g ml−1 , for quality control of Prozac
capsules
[222]
Folic acid First and second derivatives [223]
Gentamycin 78–313 g ml−1 [224]
[208]

12 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 3 (Continued )
Compounds Remarks Reference
Guanoxan sulfate First and second derivatives, in tablets, urine and serum [225]
Houttuynin sodium bisulfite First derivative, in compound cloprenaline hydrochloride [226]
Imidazole antimycotics Miconazole, clotrimazole, bifonazole and econazole were
analyzed
[227]
Indapamide First and second derivatives at 252.8 and 260.4 nm [228]
Ipratropium bromide First derivative at 254–268 nm [229]
Irbesartan First derivative at 263 nm, alone and in the presence of
hydrochlorothiazide
[230]
Isradipine First derivative at 248 nm, in the presence of its degradation
products, 5-35 mg ml−1 [231]
Lansoprazole Second derivative, 0.5–25.0 g ml−1 [232]
Linezolid Two methods: (1) first derivative with zero-crossing point and
peak to base measurement at 251.4 nm, (2) first derivative of the
ratio spectra at 263.6 nm
[233]
Loperamide hydrochloride Second derivative, in pharmaceutical formulations [234]
Loperamide hydrochloride Second derivative, in pharmaceutical formulations [235]
Losartan potassium First derivative at 234 nm, 4.00–6.00 g ml−1 [236]
Melatonine First derivative, 1.5–4.5 mg dl−1 [237]
Metomidate Third derivative, in the zero-crossing point [238]
Midazolam Second derivative at 222 nm, in the presence of maleic acid [239]
Mirtazapine First and second derivatives, 2–100 and 1–120 g ml−1 ,
respectively
[240]
Naproxen Second derivative, in the presence of its metabolite in human
plasma
[241]
Nimesulide Partition and location in egg phosphatidylcholine liposomes as
cell membrane models
[242]
Nimodipine and its photodegradation product Third derivative, evaluation of the photodegradation rate of the
nimodipine commercial specialties
[243]
Nitrofurans: nitrofurantoin, furazolidone or
furaltadone
First and second derivatives, in formulations and pig feed [244]
Olanzapine First derivative at 298 nm [245]
Olanzapine First derivative at 290.7 nm, 2.56 × 10−5 to 1.24 × 10−3 mol l−1 [246]
Omeprazole Second derivative, 0.2–40.0 g ml−1 [247]
Omeprazole First derivative at 313 nm, in aqueous solutions during stability
studies, 10–30 mg ml−1 [248]
Paracetamol First derivative, in human blood serum [249]
Pefloxacin Second derivative, in serum and pharmaceutical forms (tablets
and ampoules)
[250]
Piroxicam First derivative at 261.4 nm, 2.4–20.0 g ml−1 ,inanew
formulation (piroxicam-cyclodextrin)
[251]
Progesterone First derivative, in commercial formulations without estradiol [252]
Pyridoxine hydrochloride First derivative, at the zero-crossing wavelength at 306 nm for
the oral solution and at 308 nm for injection and capsules
[253]
4-Quinolone antibacterials Third derivative, norfloxacin, ciprofloxacin and sparfloxacin, the
method depends on the complexation of Cu(II), in formulations
and spiked human plasma and urine
[254]
Reboxetine First derivative at 285.1 nm, 8.0–40.0 g ml−1 [255]
Resorcinol Different derivative orders, in several types of pharmaceutical
preparations
[256]
Romener Third derivative, kinetic investigation of Romener in solution [257]
Saccharin Second and fourth derivatives, using zero-peak and peak-peak
methods
[258]
Salbutamol sulfate Second derivative, in the presence of skin interfering components [259]
Salicylic acid Second derivative, in aspirin or antipyretic composites [260]
Secnidazole First derivative at 296 nm, in the presence of its degradation
products, 4–30 g ml−1 [261]
Simvastatin Second derivative zero-crossing technique at 243 nm [262]
Sumatriptan succinate Three methods: first and second derivatives at 234 and 238 nm,
respectively, and first derivative of the ratio spectra at 235 nm
[263]
Sunscreens First derivative, in cosmetic formulations [264]
Sunscreens First derivative, in emulsions for beach [265]
Thiazide diuretics First derivative, using the zero-crossing technique, in the
presence of their photodecomposition products
[266]

Table 3 (Continued )
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 13
Compounds Remarks Reference
Thonzylamine hydrochloride First derivative at 494 nm, 8–20 g ml−1 , in tablets and nasal drops [267]
Timolol maleate First derivative, in ophthalmic solutions [268]
Triamcinolone acetonide First derivative at 274 nm, in ointment [269]
Trifluoperazine hydrochloride First and second derivative amplitudes at the zero-crossing point
of its sulfoxide derivative, main degradation product
[270]
Trimebutine maleate First derivative by measurement of the amplitude at 252.2 nm or
first derivative of the ratio spectrophotometry by measurement
of the amplitude at 282.4 nm, in the presence of its degradation
products
[271]
Valsartan Second derivative, peak-to-peak amplitudes at 221.6 and 231.2 nm [272]
Vitamin C Second and third derivatives, in microemulsions [273]
Vitamin E First derivative, 3.75–37.50 mg ml−1 , in Theragran formulations [274]
Yohimbine HCl First and second derivatives [275]
Acebutolol hydrochloride-nifedipine First derivative, with peak-to-base and zero-crossing
measurements methods, at 352 and 400 nm, 44–132 and
4–12 g ml−1 , respectively
[276]
Acetaminophen-ascorbic acid-aspirin First derivative, in commercial tablets [277]
Acetaminophen-caffeine-propylphenazone Fourth derivative at zero-crossing wavelengths of 256.6, 263.2
and 230.0 nm, respectively
[278]
Acetaminophen-mephenoxalone First method: first derivative—differential at 252.6 and
289.6 nm; second method: first derivative of the ratio spectra at
288.9 and 233.5 nm
[279]
Acetaminophen-phenprobamate Based on the compensation technique [280]
Acetylsalicylic-ascorbic acid First derivative using the zero-crossing method at 245 and
256 nm, 6.6 × 10−6 to 1.5 × 10−4 and 3.4 × 10−6 to 2.0 ×
10−4 mol l−1 , respectively
[281]
Acetylsalicylic acid; ascorbic acid; butylated
hydroxyanisole; caffeine; paracetamol;
phenobarbital; promethazine, salicylamide;
secobarbital
Fourth derivative, zero-crossing technique [282]
Acetylsalicylic acid-dipyridamol; acetylsalicylic
acid-glycine; betamethasone-salicylic acid;
cinnarizine-piracetam;
hydrochlorothiazide-ramipril; mebeverine-sufiride
First, second, third and fourth derivatives applying the
zero-crossing technique
Acetylsalicylic-salicylic acid Second derivative, in aspirin delayed-release tablet formulations [284]
Amiloride hydrochloride-hydrochlorothiazide First derivative of the ratio spectra at 302.5 and 285.7 nm,
respectively
[285]
Amiloride hydrochloride(I)–methylclothiazide(II) Second derivative, for I the zero-crossing technique was applied,
but for II the peak amplitude had to be corrected
[286]
2-Aminopyridine-tenoxicam Second derivative, for confirming the purity of tenoxicam in
bulk and tablets
[287]
Amlodipine—its pyridine photodegradation product Third derivative, the method could usefully be applied to routine
quality control of pharmaceutical formulations containing
amlodipine
[288]
Amoxycillin-bromhexine hydrochloride First derivative to eliminate the spectral interference from one
of the two drugs, at 278.8 and 326.2 nm
[289]
Amoxycillin-clavulanic acid First at 257.9 and 280.3 nm and second derivative at 273 and
285 nm, respectively
[290]
Analgetic mixtures Second derivative, paracetamol, propylphenazone and caffeine in
Dalivon®
[291]
Antazoline sulfate-naphazoline nitrate First derivative using the zero-crossing technique, linear up to
34gml−1 at 247.5 nm and up to 22 g ml−1 at 276.5 nm,
respectively
[292]
Antihypertensive drugs:
Use of derivative and derivative compensation techniques to
[293]
alprenolol-hydrochlorothiazide;
determine of a weakly absorbing minor component in the
hydrochlorothiazide-ramipril;
metoprolol-nifedipine
presence of a strongly absorbing major component.
Anti-inflammatory drugs:
acemethacin–indomethacin–piroxicam–tenoxicam
Second derivative and PLS model [294]
Antispasmodic drugs: mebeverine
hydrochloride-sulfiride; isopropamide
iodide-trifluoperazine hydrochloride
First and second derivatives, in commercial tablets [295]
[283]

14 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 3 (Continued )
Compounds Remarks Reference
Ascorbic acid-rutin First derivative at 258.8 and 337.4 nm, respectively [296]
Ascorbic acid-sodium salicylate-thiamine HCl First and second derivatives, zero-crossing technique, in
visalicyl tablets
[297]
Aspirin-caffeine-phenacetin Ratio spectrum derivative, in compound ACP tablet [298]
Aspirin-caffeine-phenacetin First derivative of ratio spectra and measurements of
zero-crossing method
[299]
Astemizole Astemizole-pseudoephedrine
Second derivative, 4.6–45.8 g ml
hydrochloride
−1 of astemizole; two methods:
[300]
first derivative in zero-crossing points and first derivative spectra
of their ratio spectra
Atenolol-nifedipine First derivative, at 282 and 390 nm, 25–75, 10–30 g ml−1 ,
respectively
[301]
Atenolol-amlodipine; haloperidol-trihexyphenidyl Zero-crossing point technique, in combined tablet preparations [302]
Benazepril hydrochloride-hydrochlorothiazide Second derivative at 253.6 and 282.6 nm, 14.80–33.80 and
18.50–42.20 g ml−1 , respectively
[303]
Benoxinate hydrochloride-its degradation products First derivative amplitude at 268.4 and 272.4 nm for benoxinate
in the presence of its alkali- and acid-induced degradation
products (first derivative amplitude at 307.5 nm), respectively,
the methods were used to investigate the kinetics of the acidic
and alkaline degradation processes at different temperatures
[304]
Benzocaine(I)–cetylpiridinium chloride(II) First derivative at 231.40 and 310.00 nm for (I) and at
220.70 nm for (II) using the zero-crossing method, 10–25 and
4–20 g ml−1 , respectively
[305]
Benzyl alcohol-diclofenac First and second derivatives, respectively, in injectable
formulations
[306]
Caffeine-paracetamol First and second derivatives with a Turbo-Pascal computer
program and Specord M-42, respectively, using the
zero-crossing technique
[307]
Carbinoxamine maleate-phenylpropanolamine
First derivative, applied to the study of the kinetic dissolution of
[308]
hydrochloride
pellets containing these compounds
Cefradoxil-cefotaxime Ratio-spectra first and second derivatives and zero-crossing
second derivative
[309]
Cefadroxil-cefuroxime First derivative at 267.3 and 292.5 nm, respectively, in
2–10 g ml−1 for each drug, determination of dissolution rate for
tablets and capsules containing each drug, the urinary excretion
patterns as the cumulative excreted have been calculated for
each drug
[310]
Cefoperazone-sulbactam First and second derivatives, in injectable formulations [311]
Cefsulodin-clavulanic acid First derivative using zero-crossing technique, in physiological
solutions used to prepare intravenous infusions of these antibiotics
[312]
Cephalexin-probenecid In two component solid dosage form [313]
Cephalothin-clavulanic acid First derivative with a zero-crossing technique [314]
Cetrimonium bromide(I)–lidocaine(II) Second derivative, II at 250 nm using the zero-crossing
technique, I by correction of the peak amplitude at 215 nm
[315]
Chlorpheniramine maleate-phenylephrine
Differential-derivative, zero-crossing first derivative and
[316]
hydrochloride
absorbance ratio, in nasal drops
Cilastatin sodium-imipenem First derivative, in Primaxin [317]
Cinoxacin-oxolinic acid First, second, third and fourth derivatives using peak-zero and
peak-peak methods
[318]
Cinnarizine-domperidone 5–25 and 2.5–30 g ml−1 , respectively [319]
Ciprofloxacinamide-N,N’-bishydroxymethylciprofloxacinamide
Fourth derivative, in presence of ciprofloxacin [320]
Clopamide-pindolol First derivative with a zero-crossing technique [321]
Codeine-paracetamol First derivative, 4.3 × 10−5 to 1.0 × 10−3 , 6.1 × 10−5 to 1.6 ×
10−3 mol l−1 , respectively
[322]
Cyproterone acetate-estradiol valerate First derivative of the ratio spectra [323]
Chlordiazepoxide-clinidium bromide First derivative, graphical method at 283.6 nm and zero-crossing
method at 220.8 nm, 0.740–12.0 and 0.983–21.62 mg l−1 ,
respectively
[324]
Chlordiazepoxide-clinidium bromide or
pipenzolate bromide
First derivative, in capsule and sugar-coated tablet [325]
Chlorzoxazone-paracetamol Second derivative in single, bulk mixtures and combined dosage
forms
[326]
Dapsone-pyrimethamine First derivative by zero-crossing method at 249.4 and 231.4 nm [327]

Table 3 (Continued )
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 15
Compounds Remarks Reference
Dextropropoxyphene hydrochloride-ibuprofen First derivative at 222 and 232 nm, respectively, 0–20 mg ml−1 for both drugs
[328]
Diazepam-imipramine hydrochloride Second derivative, 2–8 and 10–70: g ml−1 , respectively, applied
to pure drug mixtures and commercial preparations
[329]
Diloxanide furoate-furazolidone-tinidazole Second derivative using zero-crossing and ratio-compensation
techniques, 7.5–15, 2.5–10, 5–20: g ml−1 , respectively
[330]
Diosmin-hesperidin Zero-order and first derivative for hesperidin at 290 nm, H-point
standard addition method from 5 to 40 g ml−1 for hesperidin and
diosmin in bulk powder mixture, Daflon 500 and Dioven tablets
[331]
Dipyrone-paracetamol First derivative using zero-crossing technique, in commercial
tablets
[332]
Dipyrone-pitophenone hydrochloride Zero-crossing second and third derivatives and by ratio-spectra
first and second derivatives
[333]
Dithiocarbamate and thiuram disulfide fungicides Second derivative [334]
Enoxacin-nalidixic acid First, second, third and fourth derivatives using peak-zero and
peak-peak, 2–12 g ml−1 [335]
Erytrosine-sunset yellow First derivative, in a pharmaceutical syrup [336]
Etofylline-salbutamol Third derivative, zero-crossing technique at 303 and 233.8 nm,
respectively
[337]
Flavonoids: Chrysin-quercetin First and second derivatives, 2-20 mg ml−1 for both drug [338]
Fluvoxamine maleate-paroxetine hydrochloride First, second and third derivatives using “peak–peak” and
“peak–zero” measurements
[339]
Fosinopril-hydrochlorothiazide Fourth derivative at zero-crossing wavelengths of 217.7 and
227.9 nm, respectively
[340]
Fosinopril-hydrochlorothiazide Three methods: (1) derivative—differential in first derivative, (2)
first derivative of the ratio spectra and (3) absorbance ratio in
the zero-order spectra
[341]
Frusemide-spironolactone First derivative, in combination drug formulations [342]
Furazolidone-tinidazole First and second derivatives, in combined tablet preparations [343]
Guaiphenesin-terbutaline sulfate Second derivative by using zero-crossing [344]
Hydrocortisone-Zn-bacitracin In synthetic mixtures of these compounds in different ratios [345]
Hydrochlorothiazide-irbesartan Three methods: (1) compensation technique using ratios of the
derivative maxima or the derivative minimum, (2) first derivative
of the ratio spectra and (3) the absorbance ratio method in the
zero-order spectra
[346]
Hydrochlorothiazide-irbesartan Second derivative at the zero-crossing wavelengths at 232.7 and
230.1 nm, 1.2–2.8 and 14.4–33.6 g ml−1 , respectively
[347]
Hydrochlorothiazide-losartan potassium Two methods: (1) the amplitudes in the first derivative of the
ratio spectra at 230.423 and 238.360 nm, respectively and(2)
based on compensation technique
[348]
Hydrochlorothiazide-losartan Fourth derivative [349]
Hydrochlorothiazide-metoprolol or propranolol First and second derivatives, in combined formulations [350]
Hydrochlorothiazide-moexipril hydrochloride Second derivative with zero-crossing measurements at 234 and
215 nm, respectively
[351]
Hydrochlorothiazide-ramipril or spironolactone Vierordt’s method and ratio-spectra zero-crossing derivative [352]
Hydrochlorothiazide-triamterene Fourth and first derivatives, at 227 and 241 nm, respectively, in
combined tablets
[353]
Hydrochlorothiazide-valsartan Two methods: (1) based on compensation technique and (2)
differential first derivative
[354]
Hyoscine butylbromide-medazepam Second derivative, zero-crossing amplitude at 212.5 and
peak–trough amplitudes at 252.6 and 264.8 nm, respectively
[355]
Ibuprofen-pseudoephedrine hydrochloride First derivative at 265 and 257 nm [356]
Ibuprofen-loratadine-pseudoephedrine Second derivative zero-crossing technique [357]
Isoniazid-pyridoxine hydrochloride Derivative ratio (first derivative at 250.7 and 305.8 nm,
respectively) and differential derivative methods
[358]
Isoniazid-rifampicin First derivative UV for isoniazid and Vis spectrophotometry for
rifampicin
[359]
Lamivudine-zidovudine First derivative and first derivative of the ratio-spectra [360]
Leucovorin-methotrexate First derivative, in biological fluids [361]
Loratidine-montelukast Second derivative zero-crossing technique at 276.1 nm for
loratidine and for montelukast peak amplitude at 359.7 nm
[362]
Mefenamic acid-paracetamol First derivative, 1.8 × 10−6 to 1.6 × 10−4 , 4.1 × 10−6 to 1.4 ×
10−4 mol l−1 , respectively
[363]

16 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 3 (Continued )
Compounds Remarks Reference
Melatonin-pyridoxine Third and second derivative zero-crossing technique over
200–300 nm, respectively
[364]
Melatonin-pyridoxine First derivative of the ratio spectra method [365]
Metronidazole-norfloxacin Fourth derivative at 294.6 and 313.2 nm, 0–40 and
0–14 mg ml−1 , respectively
[366]
Naphazoline nitrate-tetramethylthionine base First derivative by zero-crossing measurements at 257.2 and
247.5 nm, respectively, in eye drops
[367]
Nicardipine-its pyridine photodegradation products Fourth derivative, in bulk and in pharmaceutics, suitable for
uniformity content anal. In pharmaceutical preparations and
stability tests in routine control
[368]
Norfloxacin-tinidazole NF at 264.2 nm which is the zero-crossing point on first
derivative of TZ, TZ at 306.2 nm which is the zero-crossing
point of NF, 0–24 and 0–36 mg ml−1 , respectively
[369]
Omeprazole-pantoprazole First derivative applying zero-crossing method for omeprazole,
omeprazole sulfone, pantoprazole sodium salt and
N-methylpantoprazole
[370]
Paracetamol-propacetamol First derivative zero-crossing wavelength at 239 and 242 nm,
respectively
[371]
Phenobarbitone-phenytoin sodium Second derivative at 244.8 and 252.8 nm, respectively,
7.5–25 g ml−1 for both compounds, in combined tablet
preparations
[372]
Phenobarbitone with: Oxyphenonium bromide and
meprobamate Paracetamol Acetylsalicylic acid
First and second derivatives applying zero-crossing technique [373]
Piroxicam-major metabolite 5-hydroxypiroxicam First derivative zero-crossing, at 343.5 and 332.5 nm, respectively,
0.5–12.0 g ml−1 for both compounds, in human plasma
[374]
Piroxicam-major metabolite 5-hydroxypiroxicam First derivative at 337.0 and 327.0 nm (zero-crossing wavelength),
linear up to 10.0 and 8.0 g ml−1 , respectively, in human plasma
[375]
Procaine hydrochloride-pyridoxine hydrochloride First and second derivatives, from sugar-coated tablets [376]
Procaine hydrochloride with: 4-Aminobenzoic
acid Benzoic acid Pyridoxine hydrochloride
Zero-crossing technique [377]
Pseudoephedrine(I)-some histamine H-1-receptor
First derivative of the ratio spectrum, I-fexofenadine, I-cetirizine
[378]
antagonist
and I-loratadine
Salbutamol-related impurities First and second derivatives for salbutamol aldehyde,
5-formylsaligenin and salbutamol ketone
[379]
Sulfadiazine-sulfamethoxazole-trimethoprim In multi-component sulfonamide reagent [380]
Sulfamethoxazole-trimethoprim Second derivative, in the presence of
hydroxypropyl-beta-cyclodextrin
[381]
Table 4
Biological analysis
Compounds Remarks Reference
Amniotic fluid First and second derivatives, evaluation of bilirubin, albumin and oxyhb [383]
Aromatic amino acids Second derivative, the solvent accessibility of tyrosine and tryptophan
residues in proteins
[384]
Aromatic amino acids First or second derivatives, of the ratios of the three aromatic amino
acid residues in peptides for a simple characterization of peptides
[385]
Aromatic amino acids At protein surface by size exclusion HPLC coupled with second derivative [386]
Barbiturates In blood, liver and urine by solid-phase extraction and UV differential
second derivative
[387]
Carboplatin First derivative, the upper detection limit was 150 g ml−1 [388]
Cerebrospinal fluid ferritin concentration First derivative, definitive angiographic detection of subarachnoid
hemorrhage compared with laboratory assessment of intra-cranial bleed
in computed tomography-negative patients
[389]
Conjugated dienes Second derivative, in lipid mixtures [390]
Conjugated dienes Second derivative, in biological samples, e.g. cod-liver oil [391]
Coumatetralyl and warfarin In blood samples by solid-phase extraction and subsequent UV-second
derivative
[392]
Coumatetralyl and warfarin In liver tissues by solid-phase extraction and subsequent UV-second
derivative
[393]

Table 4 (Continued )
C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24 17
Compounds Remarks Reference
Cyclomaltoheptaose (beta-cyclodextrin) and
hydroxyethyl-substituted beta-cyclodextrin
Inclusion complex-formation with chlorogenic acid, first derivative [394]
DDE (dichlorodiphenyldichloroethylene) Partition coefficients of DDE were determined in synthetic and native
membranes, as a function of temperature, lipid chain length, cholesterol
content and DDE concentration, by second derivative
[395]
Enolase Second derivative, at high hydrostatic pressure: an approach to the study
of macromolecular interactions
[396]
Glutathione(l-gamma-glutamyl-l-cysteinylglycine, Assay with GSH-400 method (Oxis International SA), second derivative, [397]
GSH)
in biological samples
Hemorphins Second derivative, from bovine hemoglobin hydrolyzate [398]
Human blood Optical properties of human blood as suspension of cells in aqueous
solutions of electrolytes and non-electrolytes
[399]
Indendione rodenticides In liver by solid-phase extraction and second derivative UV [400]
Indendione anticoagulant rodenticides In blood by solid-phase extraction on hydrophilic material and UV
second derivative
[401]
Lipid Second derivative, for direct measurement of lipid peroxidation in
oil-in-water emulsions using a multivariate calibration method (PLS)
[402]
Lipid Evaluation of lipid UV absorption as a parameter to measure lipid
oxidation in dark chicken meat
[403]
Milk protein Fourth derivative, milk protein in general and casein content in particular [404]
Peptides By amino acids composition [405]
Phenothiazine derivatives Second derivative, determination of partition coefficients between human
erythrocyte ghost membranes and water
[406]
Proteins Second derivative, estimation of structure and conformational stability [407]
Proteins Second derivative, simultaneous determination of fibrinogen, human
serum albumin and gamma globulin in plasma
[408]
Proteins Fourth derivative, under high pressure, I. Factors affecting the fourth
derivative Spectrum of the aromatic amino acids
[409]
Proteins Fourth derivative, under high pressure, II; application to reversible
structural changes, adrenodoxin, rnase A and methanol dehydrogenase
[410]
Proteins Pressure induced protein structural changes as sensed by fourth derivative [411]
Proteins Derivative near-UV absorption techniques for investigating protein
structure, a review with 31 refs.
[412]
Tryptophan and tyrosine Fourth derivative, in a variety of well-characterized proteins and in
proteins oxidized by N-bromosuccinimide
[413]
Tryptophan and tyrosine In soybean water soluble protein by fourth derivative [414]
Tryptophan, tyrosine and phenylalanine Second derivative [415]
Tyrosine Second derivative, 284.2 nm, study of ionization of tyrosine residues in
proteins
[416]
Tyrosine Second derivative, for the determination of the percentage of tyrosine
residues that are exposed to solvent in rabbit MM-creatine kinase
[417]
Warfarin Second derivative, in human urine and plasma [418]
3.6. Environmental analysis
The applications of derivative UV/Vis spectroscopy in water
analysis are summarized in a review with 23 references
[444].
Derivative spectrophotometry has been used for estimating
the contents of polycyclic hydrocarbons in suspended
solids and sediments of waters [445].
The significant contribution of various pesticides to
environmental pollution is of great concern. One of
the most important analytical tasks is the determination
of these compounds, their residues and metabolites
in environmental and clinical samples. The use of an
internal standard is proposed for first derivative determination
of azinphos-methyl in commercial formulations
[446].
The rapid estimation method of available nitrogen in
paddy soil was examined using the second derivative of the
above spectra [447].
UV derivative spectroscopy is investigated for its potential
in on-line control of various processes. One typical application
is emission monitoring of several pollutants such
SO2, NO, NO2, NH3 and aromatic hydrocarbons. The proposed
method gains selectivity and sensitivity by using the
first and second derivatives of the transmission spectrum
with respect to wavelength. These derivatives are generated
in an optical manner and are compared empirically for the
first time with the known numerical DS and conventional
transmission spectroscopy [448,449].
Finally, sodium dodecylbenzene sulfonate in environmental
water can be determinated by derivative spectrophotometric
method [450].

18 C. Bosch Ojeda, F. Sanchez Rojas / Analytica Chimica Acta 518 (2004) 1–24
Table 5
Food analysis
Substances Type of sample Remarks Reference
Amprolium
Veterinary premixes Second derivative at 279 nm for I and first [423]
hydrochloride(I)–ethopabate(II)
derivative at 315 nm for II
Ascorbic acid Fruits and vegetables Third derivative peak–peak amplitude [424]
Ascorbic acid Fruits, vegetables and fruit juices Second derivative, peak–baseline amplitude
at 267.5 nm
[425]
Azomicine-ornidazole Eggs and poultry food First derivative zero-crossing method [426]
Caffeine Some beverages Second derivative for cola, third derivative
for coffee and tea
[427]
Colorants Commercial food products First derivative using zero-crossing and
ratio spectra, Ponceau 4R-Carmoisine and
Ponceau 4R-Amaranth
[428]
Colorants Commercial food products Tartrazine, Amaranth and curcumin in a
micellar medium
[429]
Colorants Various food samples Ponceau 4R and tartrazine [430]
Colorants Sugar candy samples First derivative, tartrazine in the presence
of amaranth or carmoisine
[431]
Colorants Sugar confectionery products First derivative, Ponceau 4R-Sunset Yellow
and tartrazine- Sunset Yellow
[432]
Colorants Gelatin flavor pineapple; gelatin First derivative of the ratio spectra with
[433]
flavour tropical; creamy dessert measurements at zero-crossing wavelengths,
of vanilla
Tartrazine-Sunset Yellow-Ponceau 4R
Colorants Gelatin desserts First derivative, Patent Blue V-Carmoisine [434]
Colorants Instant fruit tea First derivative, Carmoisine-Ponceau 4R or
Sunset Yellow
[435]
Colorants In pure form and tablets First derivative, Sunset Yellow-erythrosine [436]
Colorants Various food samples Vierordt’s method and ratio spectra first
derivative, Tartrazine-Ponceau 4R
[437]
Colorants Food samples Vierordt’s method, ratio spectra first
derivative and first derivative,
Indigotin-Ponceau 4R
[438]
Dithiocarbamate Tomatoes Second derivative, dithiocarbamate
fungicide residues as methyl xanthate,
thiram on tomatoes
[439]
Dithiocarbamate Apple and lamb’s lettuce Second derivative dithiocarbamate
fungicide residues as methyl xanthate,
thiram on apple and lamb’s lettuce
[440]
Furfural and 5-(hydroxymethyl)-2- Locust bean extract First derivative, this compounds are useful [441]
furaldehyde
indicators of accurate storage temperature
of food samples
Saccharin Artificial sweeteners Second and fourth derivatives, in the
presence of excipients and active ingredients
[442]
Whey protein fraction mixed with
Milk Assessment of heat treatment of milk: A
[443]
casein
proposal for a new index using UV
derivative
4. Conclusions
The range of application of derivative spectrophotmetry
increases regularly in the field of analysis. DS is a relatively
modern technique which has proved to be very advantageous
in solving particular analytical problems which normal
spectroscopy is not able to solve. The advantages of the
derivative UV-Vis spectroscopy in the quantitative analysis
and the interpretation of overlapping bands are well known.
Both low noise level and relatively simplicity of the origin
UV-Vis spectra lead to a popularity of their derivatives in
spite of the signal/noise level decrease. Practically, it is not
a problem to obtain derivative spectra of any modern spectrophotometer.
The development of chemometric methods and their application
in analytical chemistry have significantly amplified
the potential power of various spectral techniques. Thus, the
precise and accurate quantification of two or more components
whose absorption bands are partly overlapped is no
longer a problem in modern spectrophotometry. Successful
resolution of binary (or more than two components) mixtures
has been obtained using full-spectrum chemometric algorithms
based on factor analysis (PCR, PLS). These methods
have been applied both to zero-order and derivative spectra,
depending on the spectral properties of the components
to be analyzed and on the type of interferences occurring
in the sample. If the components do not interact chemically
and their absorption spectra are not totally overlapped, satis-

factory resolution of the mixture can be obtained using first
derivative spectra without resorting to factor analysis.
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