Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

Review Article

ANALYTICAL, BIOCHEMICAL AND SYNTHETIC APPLICATIONS OF

Para-DIMETHYLAMINOBENZALDEHYDE

Olajire A. Adegoke
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Ibadan, Ibadan, Nigeria.

Accepted on: 24-07-2011; Finalized on: 20-11-2011.

ABSTRACT
Para-dimethylaminobenzaldehyde (Ehrlich’s reagent) has found usefulness in a wide range of applications from analytical,
biochemical to synthetic procedures and processes alongside other miscellaneous applications. The unique structural feature of
containing a para-dialkylamino substituent to an aldehyde moiety renders it highly reactive towards a wide range of compounds and
this has made it useful as a condensation reagent, oxidizable and reducible reactant in many reactions. This paper reviews the
diverse applications of DMAB spanning over a century with a prospect that this compound will be more relevant in years to come in
microbiology, chemical pathology, and organic synthesis as well as pharmaceutical and analytical chemistry. Its extensive synthetic
pathway appears to be a demerit to its application and a simpler pathway must be proposed to continue to make the compound
readily available.
Keywords: Para-dimethylaminobenzaldehyde (DMAB), Applications, Overview, Future prospects.

1.0 INTRODUCTION are aldehydes, as are several natural and synthetic

hormones2.
Aldehydes and ketones are simple compounds which
contain a carbonyl group - a carbon-oxygen double bond. Aromatic aldehydes have enjoyed a wide range of
They are simple in the sense that they don't have other applications in the development of synthetic, analytical
reactive groups like -OH or -Cl attached directly to the and biochemical processes. One of such aromatic
carbon atom in the carbonyl group1. Oxygen is far more aldehyde that has enjoyed the widest application is para-
electronegative than carbon and so has a strong tendency dimethylaminobenzaldehyde (DMAB). This paper reviews
to pull electrons in a carbon-oxygen bond towards itself. the practical applications that DMAB has been put into
One of the two pairs of electrons that make up a carbon- for over a century with a view to discovering the
oxygen double bond is even more easily pulled towards relevance of the compound and its future prospect.
the oxygen. That makes the carbon-oxygen double bond
2.0 DESCRIPTION OF DMAB
very highly polar. The slightly positive carbon atom in the
carbonyl group can be attacked by nucleophiles. A Para-Dimethylaminobenzaldehyde (I, II-3D structure) is a
nucleophile is a negatively charged ion (for example, a bifunctional aromatic skeleton possessing the aldehyde
cyanide ion, CN-), or a slightly negatively charged part of a (CHO) para to an activating substituent dimethylamino
molecule (for example, the lone pair on a nitrogen atom group [-N(CH3)2]. It‘s other synonyms are 4-
in ammonia, NH3). During the reaction, the carbon- (dimethylamino) benzaldehyde, p-(dimethylamino)-
oxygen double bond gets broken. The net effect of all this benzaldehyde, 4-Dimethylamino benzaldehyde;
is that the carbonyl group undergoes addition reactions, Ehrlichovo; Ehrlich's Reagent; p-Formyl-N,N-
often followed by the loss of a water molecule. This gives dimethylaniline; p-DAB; N,N-Dimethyl-4-amino
a reaction known as addition-elimination or benzaldehyde; 4-Dimethylaminobenzene carbonal; 4-N,N-
1
condensation . Dimethylamino benzaldehyde; N,N-Dimethyl-p-amino
benzaldehyde; N,N-Dimethyl-4-aminobenzaldehyde.
Aldehydes also undergo other wide variety of chemical
reactions, including polymerization. Their combination H3C H

with other types of molecules produces the so-called N C

aldehyde condensation polymers, which have been used H3C O

in plastics such as Bakelite and in the laminate tabletop
material Formica. Aldehydes are also useful as solvents I II
and perfume ingredients and as intermediates in the
DMAB is a white crystalline powder with a melting point
production of dyes and pharmaceuticals. Certain
of 72-75OC and boiling point of 176-177OC. Its molecular
aldehydes are involved in physiological processes.
weight is 149.19 with a molecular formula of C9H10NO and
Examples are retinal (vitamin A aldehyde), important in
log P value of 1.8. It has two hydrogen bond acceptors
human vision, and pyridoxal phosphate, one of the forms
with no hydrogen bond donor. It is stable under ordinary
of vitamin B6. Glucose and other so-called reducing sugars
condition though light sensitive.

International Journal of Pharmaceutical Sciences Review and Research Page 17

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

2.1 Synthesis of p-Dimethylaminobenzaldehyde rotation of the dimethylamino group will lead to

important volume contraction7.
The most satisfactory method for the preparation of p-
dimethylaminobenzaldehyde is the condensation of Absorption and steady-state and time-resolved emission
dimethylaniline, formaldehyde and p-nitrosodimethyl studies of DMAB in aqueous α-cyclodextrin (α-CD)
aniline, followed by hydrolysis, a method for which details solutions have been reported. The twisted intramolecular
have been perfected3. This synthetic pathway is shown in charge transfer (TICT) emission is extremely poor in pure
Figure 1. In a modification of this popular procedure, water, but is greatly enhanced upon complexation with α
Ingvaldsen and Bauman4 proposed that since an excess of -CD. The cavity size of α -CD is insufficient to encapsulate
the nitroso body is used, the second molecule of the entire fluorophore; rather it embeds DMAB only
aldehyde is partially converted into the benzylidene body, partially, keeping the dimethylamino group open in bulk
the procedure is modified in several respects. The crude water. The enhancement in the TICT emission is
aldehyde is purified by distillation in a partial vacuum. attributed to the reduced polarity provided by the CD
8
environment . The twisted motion is illustrated in Figure
2.

Figure 2: Twisting motion of DMAB in α-cyclodextrin (ref 8)

On forming semicarbazones, Trzesowska9 showed on the
Figure 1: Synthetic pathway for DMAB basis of quantum mechanical calculations the availability
2.2 Peculiar Structural features of p-Dimethylamino of the imine nitrogen atom for bonding with molecular
species thereby attesting to the wide applicability of
benzaldehyde
these derivatives of DMAB. Thus, the imine nitrogen atom
DMAB possesses some peculiar structural features which in compound of configuration E around CAN bond is likely
might account for its applicability in a wide range of not to be involved in semicarbazone–receptor
reactions and processes. 4-Dimethylaminobenzaldehyde interactions. Change of configuration leads to differences
is a para-substituted benzene derivative with a donor– in the availability of electron donor atom. The rotation
aromatic–acceptor structure. This facilitates the energy barrier is small and it is mostly related to energy
contribution of two resonance forms to the electronic of intramolecular hydrogen-bond break. The molecular
ground state (the neutral benzenoid structure and the structure of p-dimethylaminobenzaldehyde semi
zwitterionic quinonoid structure with a negative charge carbazone is shown in Figure 3.
centred on the oxygen and a positive charge centred on
5
the nitrogen) . This electronic property coupled with
three possible attachment points of DMAB to a surface
provides a complex molecule-surface system. In an
attempt to understand the DMAB-surface interaction,
gold nanoparticles were utilized6. Absorption spectra
collected for samples in aerobic and anaerobic conditions
show that gold nanoparticles in an aqueous medium in
the presence of dissolved oxygen catalyze the oxidation
of 4-(dimethylamino)benzaldehyde to 4-
(dimethylamino)benzoic acid in the pH range 2.6–11.7.
GC and GC–MS results show that at acidic pH the only Figure 3: The molecular structure of p-
dimethylaminobenzaldehyde semicarbazone. The displacement
additional product is 4-(methylamino) benzoic acid
ellipsoids are drawn at 50% probability level (ref 9)
whereas at basic pH a variety of by-products are formed.
The 15 nm diameter gold particles employed are much The heat capacities of p-dimethylaminobenzaldehyde
larger than the gold particle diameters typically used for were measured between 80 and 360 K by Meng et al 10
CO oxidation. with a small sample automated adiabatic calorimeter. The
thermodynamic parameters of solid-liquid phase
In a study of the lowest triplet of the intramolecular
transition were also obtained. The melting point, enthalpy
charge-transfer process using DMAB as a probe, the
and entropy of fusion of this compound were determined
molecule was discovered not to have a pronounced
to be 346.15 K, 19.07 kJ mol-1 and 55.08 J mol-1 K-1,
charge transfer character in apolar solvents, since the
respectively.
International Journal of Pharmaceutical Sciences Review and Research Page 18
Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

3.0 REPORTED APPLICATIONS OF The color reaction of indole alkaloids with the DMAB/iron
P-DIMETHYLAMINOBENZALDEHYDE (DMAB) (III) chloride reagent suitable for the assay of various
formulations of pharmaceuticals has been widely
Based on the above considered peculiar structural 14-20
reported .
features of DMAB, several applications have been
reported for this compound. This section of this review The spectrophotometric detection of 7-aminocephalo
report chronicles the diverse applications of this sporanic acid (7-ACA) is possible through imine formation
important derivative of benzaldehyde. of its amino functionality with DMAB 21. This kind of
derivatization was originally developed for the detection
3.1 Analytical Applications
of 6-aminopenicillic acid22, 23. It has also been applied to
24, 25
The analytical applications of DMAB comprise its the determination of 7-ACA and 7-amino
26, 27
utilization for the determination of a wide range of desacetoxycephalosporanic acid after cleavage of
substances notably among which are pharmaceuticals their respective glutaryl derivatives.
(inorganic and organic), bioactive substances and
The herbicide, isoproturon, has been determined
nanomaterials. In the applications of DMAB as an
spectrophotometrically using DMAB with good accuracy
analytical reagent, use is made of its peculiar properties
though methanolic NaOH was used for hydrolysis which
of forming condensation products, the ability of its 28
was criticized by later studies .
aldehyde moiety being reduced to the alcohol or oxidized
to the carboxylic acid. DMAB has also been used in inductively coupled plasma
mass spectrometry (ICP-MS) and inductively coupled
3.1.1 Spectrophotometric methods
plasma atomic emission spectrometry (ICP-AES) which
DMAB has been adopted solely or in combination with have been introduced and applied in analytical chemistry.
some reagents for the spectrophotometric analyses of In particular, DMAB is used in the preconcentration steps
pharmaceuticals. thus detection limits for ICP-MS and ICP-AES are at the
very low ng/L and ng/ml, respectively, which may be
A simple, accurate and sensitive spectrophotometric
useful for the determination of trace metals in real
method has been developed and validated for
samples29.
determination of H2-receptor antagonists: cimetidine,
famotidine, nizatidine and ranitidine hydrochloride. The The rate of condensation of DMAB and other aromatic
method was based on the oxidation of these drugs with aldehydes with 5-N-benzoylamino-1, 3, 4-thiadiazole-2-
cerium (IV) in presence of perchloric acid and subsequent acetonitrile has been studied and applied to the
measurement of the excess Ce (IV) by its reaction with spectrophotometric determination of these aldehydes30.
DMAB to give a red colored product (λmax at 464 nm). The
Another set of spectrophotometric applications of DMAB
decrease in the absorption intensity of the colored
include the formation of Schiff bases. The Schiff bases are
product (ΔA), due to the presence of the drug was
formed by the condensation of the aldehyde functional
correlated with its concentration in the sample solution.
with amino donors and since this extends electronic
The results obtained by the proposed method were
conjugation many of the Schiff bases are brilliantly
comparable with those obtained by the official
colored and are used for the spectrophotometric
methods11. Adegoke and Balogun also reported the
determination of the amino donors. Thus 6-APA and 6-
spectrophotometric determination of three quinolones
ACA (Figure 4) have been determined
(ciprofloxacin, sparfloxacin and perfloxacin) using Ce (IV)
spectrophotometrically in the presence of sodium
with determination of excess oxidant by a reddish-brown
dodecyl sulphate micelles via the formation of Schiff
colour formation with DMAB with good reproducibility 31, 32
12 bases . Secnidazole has also been determined
and accuracy . In both cases, the ability of excess
spectrophotometrically through Schiff base formation
unreacted Ce (IV) to oxidise DMAB is adopted.
with DMAB with measurement made at 494 nm yielding
In another application of DMAB, bopindolol was good accuracy and reproducibility33. Hydralazine has also
determined. In the proposed procedure, the been determined using Schiff base formation with DMAB
determination of bopindolol using a sequential injection with good accuracy and reproducibility. The calibration
technique (SIA) with spectrophotometric detection at 560 range obtained with the DMAB method was particularly
nm is described. The new method of determination is described as been better than previously adopted
based on the color reaction of the indole group in the methods34. The Schiff base produced with hydralazine is
molecule of bopindolol with DMAB (Ehrlich’s reagent) in shown in Figure 5. Ceftiofur has also been determined
35
acidic medium with production of a violet water-soluble through a condensation reaction with DMAB . A Schiff
complex. The selectivity of the proposed method of base formed between DMAB and sulphanilamide has
determination was tested in the presence of seven been applied as acid-base indicator which represents a
interfering substances from the group of β-blockers with major departure from previously known applications of
good results. The interference effect was observed only in Schiff bases as they are often regarded as unstable to be
36
the presence of pindolol. Obtained results were so utilized in acid-base titrations . Hydrazine derivatives
compared with conventional HPLC method both analytical supported on resin have been determined using DMAB in
13
techniques were in good agreement . alkaline media37. Benzocaine is well known as an insoluble
International Journal of Pharmaceutical Sciences Review and Research Page 19
Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

substrate in water. Therefore, its physical and analytical and diazotized nitroimidazoles. The method yielded
treatments in the presence of surfactant may allow results that were comparable with official methods and
organic solvents to be replaced by aqueous surfactants offered a promise for future applications to other
40
solutions. The effect of the presence of anionic surfactant diazotizable groups .
(sodium dodecyl sulphate, SDS) upon the condensation
DMAB has also found practical relevance in the
reaction of benzocaine with water insoluble p-
determination of environmental pollutants. The Person-
dimethylaminobenzaldehyde (DMAB) in aqueous solution
Portable Analytical Kit (PPAK) was further expanded to
has been investigated extensively. The presence of 5x10-3
allow the quantitative determination of primary amines.
M SDS increases the reaction rate by 20 times in contrast
DMAB was used to analyse for hydrazine, aniline, m-
to that of a solution in 20 % ethanol. A substantial
nitroaniline and 2, 4-diaminotoluene in tap water, waste
increase in absorption band due to the formed Schiff base
water and sea water at temperatures ranging from 10oC
followed by a red shift has been observed. A great
to 32oC41.
advantage in analytical application resulting from the
presence of surfactant due to the formation of a charge A spectrophotometric method for the determination of
transfer complex has been reported38. Chloramphenicol, total serotonin derivatives in the safflower (Carthamus
an important antibacterial agent has also been tinctorius L.) seeds has been described. The
determined via its formation of a Schiff base with determination is based upon a color reaction between
39
DMAB . serotonin derivatives and DMAB, which follows the
electrophilic substitution reaction mechanism at the
indole ring. The maximum absorption wavelength of the
complex was determined at 625 nm42. The reaction
occurring is shown in Figure 6.

Figure 4: Determination of 6-APA with DMAB via Schiff

base formation (ref 31)

Figure 6: (a)-Chemical structures of feruloylserotonin (R=OCH3)

and p-coumaroylserotonin (R=H) in safflower seeds; (b)-
condensation of tryptophan with Ehrlich’s reagent and (c)-
Reaction of indole ring derivatives with Ehrlich’s reagent (ref
42).
3.1.2 Chromatographic methods
Para-Dimethylaminobenzaldehyde has also found
relevance in some chromatographic techniques as a
derivatization reagent either in spray solutions or for
actual quantitation of molecules.
An HPTLC technique was developed for the measurement
43
of urea in pharmaceutical formulations using DMAB .
The method involves HPTLC-densitometry and
colorimetry following condensation of urea with DMAB
with better results obtained using HPTLC. DMAB as a 10 %
solution in HCl and acetone (1:4 v/v) has been used for
Figure 5: Schiff base formation between DMAB and the TLC detection of mycotoxins 44.
hydralazine (ref 34) An adaptation of the Morgan-Elson for HPLC
A recent novel application of DMAB was its utilization as a determination was proposed by Roden et al 45. The
coupling component to form azo adducts with reduced Morgan–Elson method for quantitative N-

International Journal of Pharmaceutical Sciences Review and Research Page 20

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

acetylhexosamine analysis is a two-step procedure secretion of such compounds as tryptophan, serotonin

comprising alkali treatment of the sugar and subsequent and hydroxyproline from living tissues or cell cultures.
condensation of the resulting chromogens with DMAB to This section therefore reviews the various applications of
yield a colored product. The three predominant DMAB as a biochemical tool for analysis.
components, isolated by preparative HPLC, all gave a
DMAB as presented in Kovacs’ reagent is still the reagent
purple color on addition of DMAB, indicating that they
of choice for the detection of indole produced by
were Morgan–Elson chromogens. The HPLC profile of
microorganisms and which is particularly useful for
alkali-treated N-acetylmannosamine was identical to that
identifying the Enterobacteriaceae family. Indole is
of the products generated from N-acetylglucosamine, as
generated by reductive deamination from tryptophan via
was expected because of the elimination of the
the intermediate molecule indolepyruvic acid.
asymmetry at C-2 during formation of the chromogens.
Tryptophanase catalyzes the deamination reaction,
A liquid chromatographic (LC) method for the analysis of during which the amine (-NH2) group of the tryptophan
sulfamethazine (SMT) in complete swine and cattle feed molecule is removed. Final products of the reaction are
was collaboratively studied. The method uses post- indole, pyruvic acid, ammonia (NH3) and energy. Pyridoxal
column derivatization with DMAB and detection at 450 phosphate is required as a coenzyme.
nm. The authors recommended the method for AOAC
Like many biochemical tests on bacteria, results of an
INTERNATIONAL Official First Action status46.
indole test are indicated by a change in color following a
A method is described for the HPLC determination of reaction with an added reagent. Pure bacterial culture
phenylpropanolamine (PPA) based on precolumn must be grown in sterile tryptophan or peptone broth for
derivatization with DMAB and elution from Phenomenex 24-48 hours before performing the test. Following
C-18 column with methanol–water and detection by incubation, add 5 drops of Kovacs’ reagent (isoamyl
spectrophotometry at 418 nm. Linear calibration was alcohol, p-Dimethylaminobenzaldehyde,
obtained with 9.4–46.9 µg mL−1 with a detection limit of concentrated hydrochloric acid) to the culture broth. A
4.7 ng mL−1. Vitamin B12 and rifampicin when present variation on this test using Ehrlich's reagent (using ethyl
together with PPA separated completely and could be alcohol in place of isoamyl alcohol, developed by Paul
determined simultaneously47. Ehrlich) is used when performing the test on non-
fermenters and anaerobes. A positive result is shown by
3.1.3 Spot Tests and Test Strips
the presence of a red or red-violet color in the surface
DMAB has also found usefulness in the spot tests or alcohol layer of the broth. A negative result appears
chemical functional group tests of some compounds yellow. A variable result can also occur, showing an
either in solution form or as impregnated test strips. orange color as a result. This is due to the presence
Validation procedures were described for 12 chemical of skatole, also known as methyl indole or methylated
spot tests including cobalt thiocyanate, Dille–Koppanyi, indole, another possible product of tryptophan
Duquenois–Levine, Mandelin, Marquis, nitric acid, DMAB, degradation. Indole-Positive Bacteria that test positive for
ferric chloride, Froehde, Mecke, Zwikker and Simon’s cleaving indole from tryptophan include: Aeromonas
(nitroprusside) for drugs of abuse. The validation hydrophilia, Aeromonas punctata, Bacillus alvei,
procedures include specificity and limit of detection. most Citrobacter sp., Edwardsiella sp., Escherichia coli,
Depending on the specificity of each color test, between Flavobacterium sp., Haemophilus influenzae, Klebsiella
28 to 45 drugs or chemicals were tested in triplicate with oxytoca, Proteus sp. (not P. mirabilis), Plesiomonas
each of the 12 chemical spot tests 48. Likewise, gelatinous shigelloides, Pasteurella multocida, Pasteurella
solidified layers of the photographic film were used for pneumotropica, Streptococcus faecalis and Vibrio
the immobilization of analytical reagents for detection species. Bacteria which give negative results for the
and determination of reductants and primary aromatic indole test include: Actinobacillus spp., Aeromonas
amines. It was shown, that the films with immobilized salmonicida, Alcaligenes sp., most Bacillus sp.,
iron(III)-Dipy or iron(III)-Phen complexes as test films for Bordetella sp., Enterobacter sp.,Lactobacillus spp., most
reductants and films with immobilized aldehydes (vanillin, Haemophilus sp., most Klebsiella sp., Neisseria sp.,
DMAB) as the test films for primary aromatic amines can Pasteurella haemolytica, Pasteurella ureae, Proteus
be used. The improving of reagents immobilization in the mirabilis, Pseudomonas sp., Salmonella sp., Serratia
presence of sodium dodecyl sulphate micelles was sp., Yersinia sp.50, 51 Kitasato's discovery that Escherichia
obtained. The suggested test films for the determination coli could be distinguished from Klebsiella species
of ascorbic acid, analgin (dipyrone), novocaine and (Aerobacter aerogenes) by virtue of its ability to produce
49
streptocide in drugs were examined successfully . indole initiated the detection of indole production as an
52
accepted analytical tool for the bacteriologist . The use
3.2 Biochemical Applications
of p-dimethylaminobenzaldehyde for this purpose, first
53 54
Erhlich’s reagent derived its first set of applications in applied by Bohme , modified by Kovacs , and later
55
most biochemical applications. In particular, the use of refined by Gadebusch and Gabriel , has become the
the reagent has been the age-long method for the accepted method for the detection of this metabolite in
detection of indoles by microorganisms as well as the cultures. Thus, Edwards and Ewing56 and Kauffmann57

International Journal of Pharmaceutical Sciences Review and Research Page 21

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

recommended the use of the simple Kovacs' test (p-DAB, chitinase present in the sample (Fenneropenaeus indicus
direct). The Manual of Microbiological Methods 58 agglutinin), and then detection of released GlcNAc by p-
indicates Gore's59 modification is more specific than the dimethylaminobenzaldehyde (DMAB) reagent. The
aforementioned Kovacs' test. The indole reaction is absorbance of the samples was read at 585 nm against
63
illustrated in Figure 7. the reagent blank .
Collagen integrity is often assayed by a measurement of
the content of hydroxyproline and DMAB has also found
wide applications in this regard64. The biochemical
composition and biomechanical properties of articular
cartilage from 53 human thumb carpometacarpal (CMC)
joints from cadavers aged 20 to 79 years were measured
and studied in normal, mildly fibrillated, and advanced
osteoarthritic (OA) joints Colorimetric analysis was then
used to measure the OH-Pro content when the residues
were oxidized with chloramines-T reagent to form a
65
compound that reacted with the DMAB .
Allografts of articular cartilage are both used clinically for
tissue-transplantation procedures and experimentally as
model systems to study the physiological behaviour of
chondrocytes in their native extracellular matrix. Long-
term maintenance of allograft tissue is challenging. The
Figure 7: The indole reaction by bacteria overall collagen content was assessed by measuring the
Hyaluronidase (HAase) activity has also been assessed by ortho-hydroxyproline (OHP) content via dimethylamino
the use of DMAB. The colorimetric Morgan–Elson assay benzaldehyde and chloramine T assay66.
method, which is based upon the generation of a new In measuring some common biochemical bone turnover
reducing N-acetyl-D-glucosamine terminus with each parameters in menopausal women, Sachdeva et al67 used
cleavage reaction, is most widely employed but is yet 5 % solution of DMAB in propanol to determine
insensitive. The colorimetric method was reinvestigated hydroxyproline content. Likewise the presence of DMAB-
and established the fluorimetric Morgan–Elson assay for reactive substances has been used to provide a tissue
HAase activity, with the optimized tetraborate reagent. measure of integrated hyperglycaemia over prolonged
Human serum HAase was easily characterized it along periods of time in streptozotocin-diabetic rats68.
with its optimum pH and kinetic parameters60.
DMAB has also been used as a reagent in studying many
The kynurenine (KYN) pathway of tryptophan (TRP) enzyme activities especially those related to the semi-
degradation on gene transcription of inducible nitric oxide synthesis of antibiotics. The soluble penicillin G acylase
synthase (iNOS) and nitric oxide (NO) production in (PGA) from Bacillus megaterium was used for the
chicken interferon gamma (ChIFN-γ)-stimulated and non- synthesis of cefaclor. The enzyme activity was determined
stimulated chicken macrophage cell line HD11 has also by a spectrophotometric assay with DMAB as a
been investigated spectrophotometrically using DMAB61. colorimetric substrate69.
GM2 ganglioside, β-GalNAc-(1-4)-[α-Neu5Ac-(2-3)-]β-Gal- Simple and sensitive spectrophotometric and
(1-4)-β-Glc-(1-1)-Cer, is the main ganglioside in the brain radiochemical procedures were described for the assay of
of Tay-Sachs patients. GM2 ganglioside was extracted acetyl-CoA: arylamine N-acetyltransferase (NAT: EC
from a Variant B Tay-Sachs human brain, purified to 2.3.1.5) which catalyzes the reaction acetyl CoA +
homogeneity of the oligosaccharide moiety by silica gel arylamine - N-acetylated arylamine + CoASH. The
chromatography. It was further fractionated for the first methods were applicable to crude tissue homogenates
time into the molecular species differing in the ceramide and blood lysates70.
structures by reverse-phase flash chromatography. The
GM2 ganglioside species were characterized by gas- The effective production of 7-aminocephalosporanic acid
chromatography, nuclear magnetic resonance (7-ACA) is a matter of concern in the pharmaceutical
spectroscopy, and mass spectrometry. Gangliosides on industry because it is a starting material for the synthesis
the TLC plate were made visible by spraying the plate of semi synthetic cephalosporin. Therefore screening for
with a DMAB followed by heating at 120oC for 20 min62. new source of cephalosporin acylase positive bacteria is
very important. The cephalosporin acylase can be found
A natural agglutinin from the serum of Indian white in several Pseudomonas sp. and other bacteria. To
shrimp (Fenneropenaeus indicus) was isolated and its facilitate the attempts of obtaining the microorganisms
chinitase activity was assessed using DMAB reagent. This with higher cephalosporin acylase activity from natural
assay involves release of N-acetylgalactosamine (GlcNAc) environments, development of new and specific methods
from colloidal chitin upon enzymatic hydrolysis by for screening environmental microorganisms with

International Journal of Pharmaceutical Sciences Review and Research Page 22

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

cephalosporin acylase activity is very important. For (PMSF) and tryptophan specific N-bromosuccinimide
detection of microorganisms with cephalosporin acylase (NBS) resulted in the complete inactivation of the
activity, bacteria were grown for 3 day; then each colony enzyme. DMAB method was used for kinetic
75
on the plate was exposed to chloroform vapour for 15 investigations instead of the hydroxyl amine method .
min, scraped with a toothpick, and suspended in 100 µL
The effects of the I5 polyol compounds on the
of GL-7ACA (1 mg of GL-7ACA or cephalosporin C per ml
thermostability of penicillin G acylase (PGA) from a
of 0.1 M phosphate buffer [pH 7.0]) in a well of a
mutant of Escherichia coli ATCC 11105 were investigated
microtitre dish. The mixture was incubated at 37°C for 60
by monitoring acylase activity with DMAB76.
min, and the reaction was terminated by addition of 120
µL of acetic acid : 4.25 M NaOH (2:1), followed by The effect of pHs between 2.0 and 10.0 on the
addition of 40 µL of p-dimethylaminobenzaldehyde (0.5% inactivation kinetics of penicillin G acylase (PGA) obtained
in methanol). Para-dimethylaminobenzaldehyde forms a from a mutant of Escherichia coli A TCC 11105 and the
yellow condensation product with 7-ACA or stabilization of enzyme against pH by chemical cross-
cephalosporin C71. linking with dimethyladipimidate (DMA) were studied
with enzyme activity monitored with DMAB method77.
A grapelike odour is often of diagnostic importance in
detecting the growth of Pseudomonas aeruginosa in The induction of ergot alkaloid synthesis at enzymatic
culture and in burn wounds. The compound responsible level was studied by measuring the level of tryptophan
for the odour has been identified as 2- using the DMAB method78.
aminoacetophenone (2AA) by mass spectroscopy.
3.3 Some Biomedical Applications
Although the grape odour is sometimes difficult to detect
in culture media, gas chromatographic, fluorimetric, and DMAB has also been utilized for the biomedical analysis
colorimetric methods can be utilized to assay 2- of endogenous substances. Diagnostic tests on 102 male
aminoacetophenone production in a variety of media. Its patients suspected with cannabis abuse were done.
synthesis occurs relatively early in the growth cycle. It has Liquid-liquid extraction of cannabinoids from urine was
proved easy and convenient to detect 2- done and screened by Duquenois-Levine, fast blue B salt
aminoacetophenone excretion by P. aeruginosa after 24 h and p-dimethylaminobenzaldehyde (p-DMAB) tests. All
of incubation on blood agar plates employing a the results were confirmed by high performance thin
fluorimetric assay of ether extracts of the agar medium. layer chromatography (HPTLC). Samples were considered
Thin-layer chromatograms of authentic 2AA and ether positive for cannabis based on the positive indication in
extracts of alkaline culture media yielded single, yellow colour test and by detection of 11-nor-Δ9
spots at the position of 2AA when the developed tetrahydrocannabinol-9-carboxylic acid (THC-COOH) on
chromatograms were sprayed with Ehrlich reagent (1 g of HPTLC 79. Likewise, p-amino compounds in acidic and
p-dimethylaminobenzaldehyde in a solution containing 25 ethanolic solution combine directly with DMAB to yield a
mL of HCl and 75 mL of methanol)72. yellow product. The color formation is pH sensitive, with
an optimal pH zone. This has been used for analyzing
In order to study enzyme immobilization on chitosan
biological fluids for primary arylamines and especially for
activated with glutaraldehyde, aiming to produce a cheap
the determination of p-aminohippuric acid in renal
biocatalyst, two different immobilization strategies were
function studies80. This method is a modification of
studied: one-point and multipoint covalent attachment to
previously developed method for sulphanilamide81 and p-
the solid matrix. The multipoint covalent attachment
aminohuppuric acid82 in clinical samples.
derivative had an 82% immobilization yield. Enzyme
activity was assessed via colorimetric analysis using 3.4 Synthetic Applications
73
DMAB .
DMAB has found a profound use in many synthetic
The various factors and conditions that can lead to designs as a suitable carbonyl donor and for the
production of mutant strains of microorganisms have also introduction of para-dimethylaminobenzene substituents
been studied by adopting DMAB as a reagent for into many synthetic compounds. The formation of most
monitoring one metabolite or another. Two mutants reported Schiff bases- imines, hydrazones and
have been described in which the synthesis of semicarbazones have all adopted DMAB as one of the
tryptophanase is unusually insensitive to catabolite aldehyde donors. This section of this paper reviews some
repression. Transductants were purified and grown of the synthetic applications where DMAB has found
overnight in L-broth. The addition of DMAB showed relevance in their designs.
whether or not indole (made by Tna+ but not Tna- strains)
74 3.4.1 Schiff bases
was present .
Perhaps the major synthetic application of DMAB has
The chemical modification of penicillin G acylase (PGA)
been in the formation of varied types of Schiff bases.
obtained from a mutant of Escherichia coli ATCC 11105
Schiff bases are formed by the condensation of a carbonyl
was studied in order to identify the catalytically essential
group with an amino donor. The general synthetic
amino acid residues of the enzyme. The modification of
pathway for these compounds is presented in Figure 8.
PGA by serine specific phenylmethylsulphonylfluoride

International Journal of Pharmaceutical Sciences Review and Research Page 23

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X
R
R and polymerization reactions. However, in an application
C O + H2N R"
C NR'' H2O of DMAB, dimethylaminobenzaldimine–zinc-bis
+ (pentafluorothiophenolate) was obtained from the
R'
R'
reaction of DNAB with ammonia-contaminated zinc-
When R'' = Alkyl or aryl, the product is a Schiff's base bis(pentafluorothiophenolate) and characterized by a
R'' = NH2 (hydrazine), the product is a hydrazone
structure determination. Zinc effects the catalytic
R'' = NHCONH2 (semicarbazone), the product is a semicarbazide
formation and stabilization of the aldimine by
R'' = OH (hydroxylamine), the product is an oxime
complexation89. The structures and molecular projections
Figure 8: General Synthetic Pathway for Schiff bases and
of the compounds are presented in Figure 9.
related compounds
A 1-(4-dimethylaminobenzyl)-2-(4-dimethylaminophenyl)-
benzimidazole was synthesized by the reaction of DMAB
and o-phenylenediamine. The structure was established
by X-ray crystallography and found not to be the regularly
reported rearrangement product, N, N`-bis(4-
dimethylaminobezylidene)-benzene-1,2-diamine83.
Complexes of Iron, Cobalt, Nickel and Zinc ions with the
Schiff base derived from DMAB and o-aminobenzoic acid
were synthesized and investigated by several techniques
using elemental analysis (C,H,N), molar conductance
measurements, infrared and electronic spectra. The
elemental analysis data suggest the stoichiometry to be
1:1 [M:L] ratio formation84.
Similarly, a novel N-Substituted-phenyl-1,2,3- triazole-4-
acylhydrazone was prepared by the condensation of
Figure 9: Structures and molecular projections of Zinc-
DMAB with 1-Phenyl-1H-1,2,3-triazole-4-carbohydrazide.
Aldimine, (a): ring-parallel conformers in 2a, 2b and 2c(1).
The new compound was found to have profound anti-
(b): ring-separated conformer in 2c(2) (ref 89).
platelet aggregation properties85.
3.4.2 Semicarbazones
DMAB has also been used for the synthesis of
thermotropic substances. In one of such synthesis, a new Second-order nonlinear optical (NLO) materials have been
series of Schiff base esters, 4- attracting considerable attention due to their broad
(dimethylamino)benzylidene-40-alkanoyloxyanilines applications in optoelectronics, such as optical frequency
containing even number of carbons at the end group of conversion and optical parameter oscillator. It has long
the molecules (Cn-1H2n-1COO, n = 6, 8, 10, 12, 14, 16, 18) been recognized that molecular chromophores could
were synthesized. The compounds were monotropic exhibit a second-order NLO response several orders of
liquid crystals. It was also found that the end groups of magnitude larger than that of inorganic compounds, such
the molecules had effect on the mesomorphic as potassium dihydrogen phosphate and lithium niobate.
properties86. In this regard, DMAB thiosemicarbazone was synthesised
alongside others with DMAB thiosemicarbazone showing
In another application, a series of neutral bis-ligand CuII,
the largest first-order molecular hyperpolarizabilities
NiII, PdII and PtII chelates with Schiff base ligands derived
(β)90.
from S-benzyldithiocarbazate and DMAB were prepared
and characterized. The Schiff base acts as a single Similarly, a new series of 2-[(phenylmethylene)
negatively charged bidentate ligand forming stable hydrazono]-4-oxo-3-phenyl-5 thiazolidineacetic acids
neutral metal complexes. Magnetic and spectroscopic were synthesized. Benzaldehyde 4-phenyl-3-
II II
data suggest a square-planar structure for the Ni , Pd thiosemicarbazones substituted (including DMAB) were
II
and Pt chelates. Also, ESR spectral and variable also obtained and used as intermediate to give the title
temperature magnetic susceptibility data support the compounds. All synthesized compounds were
square-planar structure of CuII chelate87. characterized by IR, 1H and 13C NMR. The in vitro anti-
Toxoplasma gondii activities of the compounds were
Arulmurugan et al recently presented a comprehensive
evaluated with IC50 ranging from 0.05-1nM91.
review of majority of Schiff bases and their metal
complexes synthesised, characterised both 3.4.3 Dye Synthesis
physicochemically and pharmacologically. Majority of
DMAB has also found application in the synthesis of
these Schiff bases are produced using DMAB with other
various dyes with different possible applications. The
amino donors88.
pyrimidinetrione system of barbituric acid may be
Aldehyde imines, the simplest of all Schiff bases, are not envisaged as a potentially useful fragment in the design of
stable at room temperature, undergoing condensation new cyanine dyes. Conjugation of the carbonyl groups of

International Journal of Pharmaceutical Sciences Review and Research Page 24

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

this ring system with a nitrogen or oxygen atom of Cyanines are a class of dyes whose chemical structure is
another fragment by means of a polymethine chain characterized by two nitrogen atoms (one of which is
should lead to new dyes with interesting spectroscopic positively charged), which are separated by a conjugated
properties. The preparation and the solvatochromic bridge formed by a carbon framework. The importance of
behaviour of two dyes, obtained by condensation of N,N`- these dyes stems from their wide use in industries for
dimethylbarbituric acid with DMAB and with 4,4`-bis(N,N- many years as spectral sensitizers for silver halide
dimethylamino)benzophenone (Michler’s ketone) have photography, in optical disks as recording media, as
been described92. The structures of the dyes and their photorefractive materials, in laser devices and even as
ionic forms in protic solvents are presented in Figure 10. anti-tumour reagents. One of the cyanine dyes was
prepared by coupling bromooctanoic acid with γ-picoline,
which was then condensed with DMAB to afford the
desired product as red solid 95.
A series of novel 4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-
a]thieno[2,3-d]pyrimidinium and 5-oxo-1,2,3,5-
tetrahydropyrrolo[2,1-b]quinazoliniumstyryl dyes were
synthesized. For preparing of studied dyes the standard
(a) method of styrylcyanines synthesis was modified.
Spectral-luminescent properties of obtained dyes in the
free state and in the presence of nucleic acids and BSA
were studied. It was shown that p-dimethylaminostyryls
based on 4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]thieno
[2,3-d]pyrimidinium with aliphatic substituents in 2 and 3
positions demonstrated RNA binding preference. These
dyes in the presence of RNA significantly enhance
(b) emission intensity and could be used as RNA-specific
Figure 10: Merocyanine-type dyes from barbituric acid fluorescent probes. Besides, the fluorescence emission
derivatives (a)-the two dyes and (b)-various ionic forms in after two-photon absorption of dye-RNA complexes in
protic solvents (ref 92). buffer solutions was measured96. One of the dyes was
The synthesis and characteristic absorption spectra of formed by a condensation reaction with DMAB.
2,3,8-trisubstituted-indeno[2,1 b]thiophene compounds 3.4.4. Miscellaneous Synthetic Applications
have been reported. Cyclization of 3-dicyanovinyl-indan-
1-one with sulphur gave 2-amino-8-oxo-8H-indeno[2,1- Para-dimethylaminobenzaldehyde has found some other
b]thiophene-3-carbo-nitrile compound. The reaction of useful synthetic applications which do not fall under the
the latter with malononitrile and with ethyl cyanoacetate categorization stated in the paragraphs above. Thus, this
gave the indeno[2,1-b]thiophene derivatives. The section reviews some other important synthetic
absorption maxima of these compounds changed very applications of DMAB.
significantly by the introduction of various substituents on Azulene and its derivatives constitute a highly interesting
the 8-position of indeno[2,1-b]thiophenes ring system. class of compounds due to the fused 5–7 bicyclic aromatic
The preparations of the corresponding azomethine dyes ring system. These compounds are regarded as one of the
and azo disperse dyes were also reported. Three representative examples of non-benzenoid aromatic
azomethine dyes were prepared by condensation hydrocarbons, which were found to be reluctant to
reaction of nitrile and thiophenes derivatives with undergo Diels–Alder reactions but easily susceptible to
DMAB93. various electrophilic substitution reactions such as
Cyclic 1,3-diones like Meldrum’s (2,2,-dimethyl-1,3- acylation, halogenation, nitration, azo-coupling and
dioxane-4,6-dione) and barbituric acid may be used as aminomethylation most easily at the C-1 and/or C-3
building blocks for intramolecular donor– acceptor pairs position. In particular, naturally occurring guaiazulene (1:
which act as solvatochromic dyes. Thus, condensation of 1,4-dimethyl-7-isopropylazulene) possesses low oxidation
DMAB with N,N-dimethylbarbituric acid leads to the potential (Eox +0.65 V vs SCE) in comparison with those of
formation of merocyanine. In this regard, The preparation azulene (Eox+0.88 V vs SCE) and other alkylazulenes (Eox
of the 5-(4-N,N-dimethylaminobenzylidene)-2,2-dimethyl- +0.88– +0.67 V vs SCE), and is an interesting compound
1,3-dioxane-4,6-dione, a dye derived from Meldrum’s from the viewpoint of the creation of novel functional
acid, was described, and its solvatochromic behaviour materials, with a delocalized p-electron system
3
compared with that of an analogous derivative of possessing a 3-guaiazulenyl group (Gu ), which serves as
94
barbituric acid . The dye was particularly found to be an electron donor. Thus, reaction of guaiazulene with
sensitive both to the dipolarity–polarizability and the DMAB in methanol in the presence of tetrafluoroboric
acidity of the medium. acid gives the title mono-carbocation compound, [4-
(dimethylamino) phenyl] -3-guaiazulenyl methylium
tetrafluoro borate, in 90% yield 97.

International Journal of Pharmaceutical Sciences Review and Research Page 25

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

Search for new materials with high optical nonlinearities 4.0 MISCELLANEOUS APPLICATIONS
has been the important task because of their practical
This part of this review reports some other applications
application in harmonic generation, amplitude and phase
where DMAB has been utilized either as a synthetic
modulation, switching and other signal processing
precursor or reagent for analysis. The formation of an
devices. In this regard, an organic nonlinear optical
alcohol from the reaction of DMAB with pyrrole
material, 4-chloro-4-dimethylamino-benzylidene aniline
derivatives having intact CH-group in the α or β position
(CDMABA), was synthesized by the condensation of the p-
relative to the cyclic NH-group have been reported and
chloroaniline and p-dimethylaminobenzaldehyde.
DMAB reagent was utilised for the spectrophotometric
Solubility of CDMABA was determined in acetone at
determination of 2-phenylindole by Gillio-Tos et al105. In a
different temperatures. Single crystals were grown by the 106
similar procedure, Adegoke and Osoye found that
solvent evaporation method from acetone solution at
reactive methylene centres generated by artemisinin
room temperature. Grown crystal was subjected to FTIR,
1 derivatives (artesunate and dihydroartemisinin) in situ in
FTRaman and H NMR spectral analyses to confirm the
an acidic medium produced a purple-coloured solution
synthesized compound. The range and percentage of
which was used for the full colorimetric determination of
optical transmission was ascertained by recording UV–
98 these important antimalarial agents.
Vis–NIR spectrum .
DMAB has also been used for the histochemical
A novel and simple fluorophore, p-
demonstration of tryptophan and related compounds107.
dimethylaminobenzaldehyde thiosemicarbazone
(DMABTS), was prepared in order to find available
5.0 OVERVIEW OF THE UTILITY STATUS AND FUTURE
fluorescent chemosensor for mercuric ion in aqueous
PROSPECTS
solution. DMABTS emitted fluorescence at 448 nm in
aqueous solution and its fluorescence intensity was This paper has made attempt to review the many diverse
completely quenched upon interaction with Hg2+ ions, applications of DMAB which spanned over a century.
which should be attributed to the 1:1 complex formation DMAB because of its peculiar structural features has
between DMABTS and Hg2+. The coexistence of several found applications in synthetic, biochemical, biomedical
transition metal ions and anions did interfere with the and analytical sciences. Its use in these respects is
fluorimetric titration of Hg2+ ion by less than 4 % in the gradually increasing and some of the age-long
emission change99. applications of this reagent are experiencing some new
modifications. The relevance of this compound in
Hybrid mixed metal oxalates of general formula [M2III
microbiology and analytical chemistry has been
MII(C2O4)6][DAMS]4.2DAMBA. 2H2O, where MIII = Rh, Fe,
unparalled by any other reagent in history. It behoves to
Cr; MII = Mn, Zn; DAMBA = para-dimethylamino
opine that the relative lack of toxicity of this compound
benzaldehyde and [DAMS+] = trans-4-(4- dimethylamino
has made it more useful in these fields. However, the
styryl)-1-methylpyridinium, belong to a new family of
synthesis of DMAB appears unnecessarily too lengthy and
multifunctional materials displaying both very high
one believes that the challenge for synthetic chemists will
second harmonic generation (SHG) efficiency and tunable
be to devise some simpler pathways for its synthesis. The
magnetic properties. Cariati et al reported the
future prospect of this reagent is still bright as its unique
preparation and magnetic characterization of the new NiII
properties of being oxidizable and reducible will lend it to
members of this family 100.
more usefulness. When a need for a dimethylamino
DMAB has also found practical relevance in the cross- substituted benzene molecule is arises DMAB comes
101
coupling reaction of arylbromides and chlorides , handy and useful. Its ability to form condensation
reductive C-alkylation of barbituric acid derivatives in the products readily with a wide range of chemical groupings
102
presence of palladium and platinum catalysts as well as such as amines, carboxylates and other aldehydes will
103
in the reductive mono-alkylation of nitroaryls . make it more relevant in synthetic design and processes.
Its recent application as a coupling component in our
Oniţa et al recently reported the synthesis of
laboratory for the spectrophotometric determination of
Aminophosphonates as potential herbicides for which
diazotized nitroimidazoles has further opened up another
DMAB served as one of the precursors. The antioxidant
area of research into its utilization for which ready
activity was tested in experimental assays with DPPH•
applications in the third world economies will be possible.
(1,1-diphenyl-2-picryl-hydrazyl) and the potential to
The reaction also has a potential of being used as a pre-
function as herbicides, in red blood cells membrane
column derivatization procedure and for which analysis of
destruction experiments. Regarding the reaction with
pharmaceuticals in dosage forms and biological fluids will
DPPH•, at least 3 aminophosphonates are better
be possible.
antioxidants than 2,6-di-tert-butyl-4-methylphenol (BHT).
Aminophosphonates’ capacity to function as herbicide
(demonstrated through their haemolytic activity) is given
by the presence in their structure of some shorter
104
aliphatic substituents and / or more benzene nuclei .

International Journal of Pharmaceutical Sciences Review and Research Page 26

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X

REFERENCES 21. Patett F, Fischer L, Spectrophotometric assay for quantitative

determination of 7-aminocephalosporanic acid from direct
1. An introduction to Aldehydes and ketones, hydrolysis of cephalosporin C, Analytical Biochemistry 350, 2006,
http://www.chemguide.co.uk/organicprops/carbonyls/background 304–306.
rd
.html#top (Date accessed 23 July 2011)
22. Balasingham K, Warburton D, Lilly MD, The isolation and kinetics of
2. Aldehydes, www.britannica.com/EBchecked/topic/13527/ penicillin amidase from Escherichia coli, Biochim. Biophys. Acta
rd
aldehyde/277601/Properties-of-aldehydes (Date accessed 23 July 276, 1972, 250–257.
2011).
23. Bomstein J, Evans WG, Automated colorimetric determination of
3. Ullmann and Frey, Ber. 37, 858,1904; Organic Syntheses, Coll. Vol. 6- aminopenicillanic acid in fermentation media, Anal. Chem. 37,
1, p.214 (1941); Vol. 2, p.17 (1922). 1965, 576–578.
http://www.orgsynth.org/orgsyn/prep.asp?prep=cv1p0214 (date
th 24. Matsuda A, Komatsu K-I, Molecular cloning and structure of the
accessed 20 July 2011).
gene for 7β-(4-carboxybutanamido)cephalosporanic acid acylase
4. Ingvaldsen T, Bauman L, Note on the preparation of para- from a Pseudomonas strain, J. Bacteriol. 163,1985, 1222–1228.
Dimethylaminobenzaldehyde, The Journal of Biological Chemistry,
25. Sonawane VC, Jolly RS, Vohra RM, Cephalosporin modification: an
XLI (2), 1919, 1-2.
extracellular glutaryl-7-ACA-acylase from Bacillus sp., Biotechnol.
5. Rosencrance JG, Jagodzinski PW, Infrared and Raman spectra and Lett. 18, 1996, 965–968.
vibrational assignments for 4-(dimethylamino) benzaldehyde and
26. Lee YS, Park SS, Two-step autocatalytic processing of the glutaryl 7-
its zinc complex, Spectrochim. Acta 42A, 1986, 869-879.
aminocephalosporanic acid acylase from Pseudomonas sp. Strain
6. Krieger RM, Jagodzinski PW, Catalytic oxidation of 4- GK16, J. Bacteriol. 180, 1998, 4576–4582.
(dimethylamino)benzaldehyde by gold nanoparticles. Part I:
27. Shibuya Y, Matsumoto K, Fuji T, Isolation and properties of 7-(4-
Reaction characterization, Journal of molecular structure 876,
carboxybutanamido)cephalosporanic acid acylase-producing
2008, 56-63.
bacteria, Agric. Biol. Chem. 45, 1981, 1561–1567.
7. Chattopadhyay N, Van der Auweraer M, De Schryver FC,
28. Raju KR, Parthasarthy TN, Akella SRKM, Spectrophotometric
Determination of the nature of the lowest triplet state of the
determination of isoproturon or metoxuron using p-
intramolecular charge-transfer probes DMABN and DMABA by
dimethylaminobenzaldehyde, Analyst 115, 1990, 455–457.
laser-induced optoacoustic spectroscopy, Chemical Physics Letters
279, 1997, 303-308. 29. Cui Y, Chang X, Zhai Y, Zhu X, Zheng H, Lian N, ICP-AES
determination of trace elements after preconcentrated with p-
8. 8. Kundu S, Chattopadhyay N, Twisted intramolecular charge
dimethylaminobenzaldehyde-modified nanometer SiO2 from
transfer of dimethylaminobenzaldehyde in c -cyclodextrin cavity,
sample solution, Microchemical Journal 83, 2006, 35-41.
Journal of Molecular Structure 344, 1995, 151-155.
30. Mediena HAA, Erian AW, Kinetic studies of the condensation of
9. Trzesowska A, P-Dimethylaminobenzaldehyde semicarbazone: The
aromatic aldehydes with 5-N-benzoylamino-1,3,4-thiadiazole-2-
bonding abilities of imine nitrogen atom, Journal of Molecular
acetonitrile, and their spectrophotometric determination
Structure 917, 2009, 125-132.
Microchemical Journal 65, 2000,31-38
10. He Meng S, Liang P, Tan ZC, Ji Song Y, Lia L, Wang L, Heat capacity
31. Yatsimirskaya NT, Sosnovskaya IN, Yatsimirisky AK,
and thermodynamic properties of p-dimethylaminobenzaldehyde,
Spectrophotometric determination of 6-Aminopenicillanic and 7-
Thermochimica Acta 342, 199, 47-51.
Aminocephalosporanic Acids as the Schiff bases with para-
11. Darwish IA, Hussein SA, Mahmoud AM, Hassan AI, Dimethylaminobenzaldehyde in the presence of sodium dodecyl
Spectrophotometric determination of H2-receptor antagonists via sulphate micelles, Analytical Biochemistry 229, 1995, 249-255.
their oxidation with cerium(IV), Spectrochimica Acta Part A 69,
32. Shewale JG, Kumar KK, Ambekar GR, Evaluation of determination
2008, 33–40.
of 6-aminopenicillane acid by p-dimethylaminobenzaldehyde.
12. Adegoke OA, Balogun BB, Spectrophotometric Determination of Biotechnol Tech 1, 1987, 69–72.
Some Quinolones Antibiotics following Oxidation with Cerium
33. El Walily AFM, Abdine HH, Razak OA, Zamel S, Spectrophotometric
Sulphate, International Journal of Pharmaceutical Sciences Review
and HPLC determination of secnidazole in pharmaceutical tablets,
and Research 4(3), 2010,1-10.
Journal of Pharmaceutical and Biomedical Analysis 22, 2000, 887-
13. Satınsky D, Sklenarova H, Huclova´ J, Karlıcek R, Determination of 897.
bopindolol by sequential injection technique with
34. Adegoke OA, Nwoke CE, Spectrophotometric Determination of
spectrophotometric detection IL Farmaco 58,2005, 1057-1062.
Hydralazine using p-Dimethylaminobenzaldehyde, Journal of the
14. Pesez M, Bartos J, Colorimetric and Fluorimetric Analysis of Iranian Chemical Society, 5 (2), 2008, 316-323.
Organic Compounds and Drugs. Marcel Dekker, New York, 1974.
35. Annapurna V, Jyothi G, Rambabu C, Sailaja BBV,
15. Kakac B, Vejdelek ZJ Handbuch der Kolorimetrie, I, II (Kolorimetrie Spectrophotometric Determination of Ceftiofur Hydrochloride
in der Biologie, Biochemie und Medizin), Gustav Fischer Verlag, Using N-Bromosuccinimide and p-Dimethylaminobenzaldehyde. E-
Jena, 1962–1966. Journal of Chemistry 6(3), 2009, 763-769.
16. Kakac B, Vejdelek ZJ, Handbuch der photometrischen Analyse, 36. 36. Khalil RA, Jalil AH, Abd-Alrazzak AY, Application of a Schiff Base
Organische Verbindungen. Verlag Chemie, Weinheim, 1977. Derived from Sulfanilamide as an Acid-Base Indicator, J. Iran.
17. Vejdelek ZJ, Kakac B, Farbreaktionen in der Chem. Soc. 6(2), 2009, 345-352
spektrophotometrischen Analyse organischer Verbindungen, I, II. 37. Gaggini F, Porcheddu A, Reginato G, Rodriquez M, Taddei M,
Gustav Fischer Verlag, Jena (1980–1982) Ergänzungsband I, II, Colorimetric Tools for Solid-Phase Organic Synthesis, J. Comb.
1969–1973. Chem. 6, 2004, 805-810.
18. Pesez M, Poirier P, Bartos J, Pratique de l’Analyse Organique 38. Khalil RA, Hussain SA, Surfactant Enhanced Reaction between
Colorimetrique, Masson, Paris, 1966. Benzocaine and p-Dimethylaminobenzaldehyde: Kinetic Study and
19. Snell FD, Snell CT, Snell CA, Colorimetric Methods of Analysis, III, IV Its Analytical Application,The Arabian Journal for Science and
(Organic Compounds), IIIA, IVA, van Nostrand, Princeton, 1948- Engineering, Volume 35 (2A), 2010, 55-66.
1970. 39. Eboka CJ, Smart J, Adelusi SA, An alternative colorimetric method
20. Görög S, Ultraviolet-Visible Spectrophotometry in Pharmaceutical for the determination of chloramphenicol, Trop. Journal of
Analysis. CRC Press, Boca Raton, 1995, pp 235–240, 190–194, Pharmaceutical Research 2(2), 2003, 215-221.

International Journal of Pharmaceutical Sciences Review and Research Page 27

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X
40. Adegoke OA, Umoh OE , A new approach to the macrophage cells upon treatment with interferon gamma,
spectrophotometric determination of metronidazole and Immunology Letters 115, 2008, 153-159.
tinidazole using p-dimethylaminobenzaldehyde, Acta
62. Mauri L, Valsecchi M, Casellato R, Su-Chen Li, Yu-Teh Li, Sonnino S,
Pharmaceutica 59 (4), 2009, 407-419.
Procedure for separation of GM2 ganglioside species with different
41. Kasitu GC, Bissonnette MC, Goldthorp M, Fingas MF, Bklanger JMR, ceramide structures by a flash reversed-phase silica gel liquid
Pare JRJ, Expansion of the capabilities of the Person-Portable chromatography, Journal of Chromatography B 796, 2003,1-10.
Analytical Kit (PPAK), Journal of Hazardous Materials 43, 1995, 63. Maheswari R, Mullainadhan P, Arumugam M, Isolation and
129-139.
characterization of an acetyl group-recognizing agglutinin from the
42. Jin Q, Shan L, Yue J, Wang X, Spectrophotometric determination of serum of the Indian white shrimp Fenneropenaeus indicus,
total serotonin derivatives in the safflower seeds with Ehrlich’s Archives of Biochemistry and Biophysics 402, 2002, 65-76.
reagent and the underlying color reaction mechanism, Food
64. Reddy GK, Enwemeka CS, A Simplified method for the analysis of
Chemistry 108, 2008, 779-783.
hydroxyproline in biological tissues, Clinical Biochemistry 29,
43. Knorst MT, Neubert R, Wohlrab W, Analytical Methods for 2002,225-229.
measuring Urea in Pharmaceutical formulations, Journal of
65. Rivers PA, Rosenwasser MP, Mow VC, Pawluk RJ, Strauch RJ,
Pharmaceutical and Biomedical Applications 15, 2008,1627-1632. Sugalski MT, Ateshian GA, Osteoarthritic Changes in the
44. Lin L, Zhang J, Wang P, Wang Y, Chen J, Thin-layer chromatography Biochemical Composition of Thumb Carpometacarpal Joint
of mycotoxins and comparison with other chromatographic Cartilage and Correlation with Biomechanical Properties, The
methods, Journal of Chromatography A 815, 1998, 3-20. Journal of Hand Surgery Vol. 25A (5), 2000, 889-898.
45. Roden L, Yu H, Jin J, Ekborg G, Estock A, Krishna NR, Livant P, 66. Bian L, Lima EG, Angione SL, Ng KW, Williams DY, Xu D, Stoker AM,
Analysis of the Morgan–Elson Chromogens by High-Performance Cook JL, Ateshian GA, Hung CT, Mechanical and biochemical
Liquid Chromatography, Analytical Biochemistry 254, 1997, 240- characterization of cartilage explants in serum-free culture, Journal
248. of Biomechanics 41, 2008, 1153-1159.
46. Smallidge RL, Albert K, Determination of Sulfamethazine in Swine 67. Sachdeva A, Seth S, Khosla AH, Sachdeva S, Study of some
and Cattle Feed by Reversed-Phase Liquid Chromatography with Common Biochemical Bone Turnover Markers in Postmenopausal
Post-Column Derivatization: Collaborative Study, Journal of AOAC Women, Indian Journal of Clinical Biochemistry 20 (1), 2005, 131-
International 83 (2), 2000, 260-268. 134.
47. Rind FMA, Khuhawar MY, Rajper AD, HPLC determination of 68. Stefek M, Gajdosik A, Gajdosikova A, Krizanova L, p-
phenylpropanolamine in pharmaceutical preparations using 4- Dimethylaminobenzaldehyde-reactive substances in tail tendon
dimethylaminobenzaldehyde as a derivatizing reagent, Journal of collagen of streptozotocin-diabetic rats: temporal relation to
Pharmaceutical and Biomedical Analysis 26, 2001, 331-336. biomechanical properties and advanced glycation end product
(AGE)-related fluorescence, Biochimica et Biophysica Acta 1502,
48. O’Neal CL, Crouch DJ, Fatah AA, Validation of twelve chemical spot
2000, 398-404.
tests for the detection of drugs of abuse, Forensic Science
International 109, 2000, 189-201. 69. Yang L, Wei D-Z, Enhanced enzymatic synthesis of a semi-synthetic
cephalosporin, cefaclor, with in situ product removal,
49. Loginova LP, Yu Konovalova O, Test films for test-determinations
on the base of reagents, immobilized in gelatinous gel, Talanta 77, Biotechnology Letters 25, 2003, 1195-1198.
2008, 915-923. 70. Andres HH, Klem AJ, Szabo SM, Weber WW, New
Spectrophotometric and Radiochemical Assays for Acetyl-CoA:
50. MacFaddin JF, Biochemical Tests for Identification of Medical
Bacteria, Williams & Wilkins, 1980, pp 173 - 183. Arylamine N-Acetyltransferase Applicable to a Variety of
Arylamines, Analytical Biochemistry 145, 1985, 367-375.
51. Isenberg HD, Sundheim LH, Indole reactions in Bacteria, The
Journal of Biological Chemistry 75, 1958, 682-690. 71. Tanomand A, Abeshov R, Farajnia S, Determination of
cephalosporin acylase activity by biological and colorimetric
52. Kitasato S, Die negative Indolreaktion der Typhusbazillen im method in bacteria, African Journal of Biotechnology 8 (23), 2009,
Gegensatz zu anderen Bazillenarten. Z. Hyg. Infektionskrankh., 7, 6697-6699.
1889, 515-525.
72. Cox CD, Parker J, Use of 2-Aminoacetophenone Production in
53. Bohme A, Die Anwendung der Ehrlichschen Indolreaktion fur Identification of Pseudomonas aeruginosa, Journal of Clinical
bacteriologische Zwecke. Centr. Bakteriol. Parasitenk. I. Abt., 40, Microbiology 9 (4), 1979, 479-484.
1906, 133-192.
73. Adriano WS, Filho EHC, Silva JA, Giordano RLC, Gonçalves LRB,
54. Kovacs N, Eine vereinfachte Methode zum Nachweis der Stabilization of Penicillin G Acylase by Immobilization on
Indolbildung durch Bacterien. Z. Immunitatsforsch., 55, 1928, 311- Glutaraldehyde-Activated Chitosan, Brazilian Journal of Chemical
315. Engineering 22 (4), 2005, 529-238.
55. Gadebusch HH, Gabriel S, Modified stable Kovacs reagent for the 74. Yudkin MD, Mutations in Escherichia coli that Relieve Catabolite
detection of indol. Am. J. Clin. Pathol., 26, 1956, 1373-1375. Repression of Tryptophanase Synthesis. Mutations Distant from
56. Edwards PR, Ewing WH, Identification of enterobacteriaceae. the Tryptophanase Gene, Journal of General Microbiology 92,
Burgess Publishing Co., Minneapolis. 1955. 1976, 125-132.

57. Kauffmann F, Enterobacteriaceae, Ed. 2. Ejnar Munksgaard, 75. Kazan D, Erarslan A, Identification of catalytically essential amino
Copenhagen, Denmark, 1954. acid residues of penicillin G acylase obtained from a mutant of
Escherichia coli ATCC 11105, Process Biochemistry 36, 2001, 861-
58. Manual of microbiological methods, Society of American 867.
Bacteriologists. McGraw-Hill Book Company, Inc., New York, 1957.
76. Erarslan A, The Effect of Polyol Compounds on the Thermostability
59. 59. Gore SN, The cotton wool plug test for indole, Indian J. Med. of Penicillin G Acylase from a Mutant of Escherichia coli ATCC
Research 8, 1921, 505-507. 11105, Process Biochemistry 30 (2), 1995, 133-139.
60. Takahashi T, Ikegami-Kawai M, Okuda R, Suzuki K, A fluorimetric 77. Kazan D, Ertan H, Erarslan A, Stabilization of Penicillin G Acylase
Morgan–Elson assay method for hyaluronidase activity, Analytical Against pH by Chemical Cross-Linking, Process Biochemistry 31(2),
Biochemistry 322, 2003, 257-263. 1996, 135-140.
61. Kujundzic RN, Lowenthal JW, The role of tryptophan metabolism in 78. Krupinski VM, Robbers JE, Floss HG, Physiological Study of Ergot:
iNOS transcription and nitric oxide production by chicken Induction of Alkaloid Synthesis by Tryptophan at the Enzymatic
Level, Journal of Bacteriology 125 (1), 1976, 158-165.

International Journal of Pharmaceutical Sciences Review and Research Page 28

Available online at www.globalresearchonline.net
Volume 11, Issue 2, November – December 2011; Article-004 ISSN 0976 – 044X
79. Sharma P, Bharath MMS, Murthy P, Qualitative high performance 93. Fu TL, Wang IJ, Synthesis and substituent effects of some novel
thin layer chromatography (HPTLC) analysis of cannabinoids in dyes derived from indeno[2,1-b]thiophene compounds, Dyes and
urine samples of Cannabis abusers, Indian J Med Res 132, Pigments 76, 2008, 158-164.
2010,201-208.
94. Flores P, Rezende MC, Jara F, A solvatochromic derivative of
80. Waugh WH, Beall PT, Simplified measurement of p- Meldrum’s acid Dyes and Pigments 62, 2004, 277-281.
aminohippurate and other arylamines in plasma and urine, Kidney
95. Mishra A, Newkome GR, Moorefield CN, Godınez LA, Synthesis,
International 5, 1974,429-436. spectroscopic and electrochemical investigation of some new
81. Morris CJO, The determination of sulphanilamide and its stilbazolium dyes, Dyes and Pigments 58, 2003, 227-237.
derivatives, Biochem J 35, 1941, 952—959.
96. Balanda AO, Volkova KD, Kovalska VB, Yu M, Losytskyy, Tokar VP,
82. Brun C, A rapid method for the determination of para- Prokopets VM, Yarmoluk SM, Synthesis and spectral-luminescent
aminohippuric acid in kidney function tests. J. Lab. Clin. Med. 37, studies of novel 4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]thieno[2,3-
1951, 955—958. d]pyrimidiniumstyryls as fluorescent dyes for biomolecules
detection, Dyes and Pigments 75, 2007,25-31.
83. Sheikhshoaie I, Belaj F, Fabian WMF, 1-(4-Dimethylaminobenzyl)-2-
(4-dimethylaminophenyl)-benzimidazole: Synthesis, X-ray 97. Sasaki M, Nakamura M, Uriu T, Takekuma H, Minematsu T,
crystallography and density functional theory calculations, Journal Yoshihara M, Takekuma S, A facile preparation, the crystal
of Molecular Structure 794, 2006, 244-250. structure, the chemical and electrochemical properties of [4-
84. Ben-saber SM, Maihub AA, Hudere SS, El-ajaily MM, Complexation (dimethylamino) phenyl]-3- guaiazulenyl methylium
tetrafluoroborate, Tetrahedron 59, 2003, 505-516.
behaviour of Schiff base toward transition metal ions,
Microchemical Journal 81, 2005, 191-194. 98. Leel S, Ramamurthia K, Bhagavannarayana G, Synthesis, growth,
spectral, thermal, mechanical and optical properties of 4-chloro-4`-
85. Cunha AC, Figueiredo JM, Tributino JLM, Miranda ALP, Castro HC,
dimethylamino-benzylidene aniline crystal: A third order nonlinear
Zingali RB, Fraga CAM, de Souza MCBV, Ferreira VF, Barreiro EJ,
Anti-platelet Properties of Novel N-Substituted-phenyl-1,2,3- optical material, Spectrochimica Acta Part A 74, 2009, 78-83.
triazole-4-acylhydrazone Derivatives, Bioorganic & Medicinal 99. Yu Y, Lin Li-R, Yang Kai-B, Zhong X, Huang Rong-B, Zheng Lan-S, p-
Chemistry 11, 2003, 2051–2059. Dimethylaminobenzaldehyde thiosemicarbazone: A simple novel
86. Ha ST, Ong LK, Ong ST, Yeap GY, Wong JPW, Koh TM, Lin HC, selective and sensitive fluorescent sensor for mercury(II) in
aqueous solution, Talanta 69, 2006,103-106.
Synthesis and mesomorphic properties of new Schiff base esters
with different alkyl chains, Chinese Chemical Letters 20, 2009, 767- 100. Cariati E, Macchi R, Tordin E, Ugo R, Bogani L, Caneschi A, Macchi
770. P, Casati N, Sironi A, Tuning the magnetic properties of a new
family of hybrid mixed metal oxalates having 1D magnetic chains
87. Tian YP, Duan CY, Lu ZL, You XZ, Fun HK, Kandasamy S, Crystal
Structure and Spectroscopic studies on metal complexes and layers of J aggregates of [DAMS+] producing superior SHG,
Inorganica Chimica Acta 361, 2008, 4004-4011.
containing NS donor ligands derived from S-benzyldithiocarbazate
and p-dimethylaminobenzaldehyde, Polyhedron 15 (13), 1996, 101. Ozdemir I, Demir S, Cetinkaya B, Use of tetrahydropyrimidinium
2263-2271. salts for highly efficient palladium-catalyzed cross-coupling
reactions of aryl bromides and chlorides, Tetrahedron 61, 2005,
88. Arulmurugan S, Kavitha HP, Venkatraman BR, Biological Activities
of Schiff Base and Its Complexes: A Review, Rasayan J. Chem. 3 (3), 9791-9798.
2010, 385-410. 102. Jursic BS, Neumann DM, Reductive C-alkylation of barbituric acid
derivatives with carbonyl compounds in the presence of platinum
89. Muller B, Vahrenkamp H, A Zinc–aldimine complex, Inorganica
Chimica Acta 300-302, 2000, 181-185. and palladium catalysts, Tetrahedron Letters 42, 2001, 4103-4107.
103. Sydnes MO, Kuse M, Isobe M, Reductive monoalkylation of nitro
90. Ren P, Liu T, Qin J, Chen C, A new approach to suppress
nonlinearity-transparency trade-off through coordination aryls in one-pot, Tetrahedron 64 2008, 6406-6414.
chemistry: syntheses and spectroscopic study on second-order 104. Oniţa N, Şişu I, Penescu M, Purcarea VL, Kurunczi L, Synthesis,
nonlinear optical properties of a series of square-pyramidal zinc(II) Characterization and Biological Activity of Some -
complexes, Spectrochimica Acta Part A 59, 2003, 1095-1101. Aminophosphonates, Farmacia 58 (5), 2010, 531-545.
91. De Aquino TM, Liesen AP, da Silva REA, Lima VT, Carvalho CS, de 105. Gillio-Tos MV, Previtera SA, Goodman EM, Spectrophotometric
Faria AR, de Araujo JM, de Lima JG, Alves AJ, de Melo EJT, Goes determination of 2-phenylindole with p-
AJS, Synthesis, anti-Toxoplasma gondii and antimicrobial activities dimethylaminobenzaldehyde, Analytical Chemistry 36 (2), 1964,
of benzaldehyde 4-phenyl-3-thiosemicarbazones and 2- 425-426.
[(phenylmethylene)hydrazono]-4-oxo-3-phenyl- 5-
106. Adegoke OA, Osoye AO, Derivatization of Artesunate and
thiazolidineacetic acids, Bioorganic & Medicinal Chemistry 16,
Dihydroartemisinin for Colorimetric analysis using p-
2008, 446-456. dimethylaminobenzaldehyde, Eurasian Journal of Analytical
92. Rezende MC, Campodonico P, Abuin E, Kossanyi J, Merocyanine- Chemistry 6, 2011, (In Press).
type dyes from barbituric acid derivatives, Spectrochimica Acta
107. Adams CWM, A p-Dimethylaminobenzaldehyde-nitrite Method for
Part A 57, 2001, 1183-1190. the histochemical demonstration of Tryptophane and related
compounds, J. Clin. Path. 10, 1957, 56-62.

*********************

International Journal of Pharmaceutical Sciences Review and Research Page 29

Available online at www.globalresearchonline.net