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SURFACTANT ENHANCED REACTION BETWEEN BENZOCAINE AND P-

DIMETHYLAMINOBENZALDEHYDE: KINETIC STUDY AND ITS ANALYTICAL
APPLICATION

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 ‫‪Rabah A. Khalil and Sana A. Hussain‬‬

‫‪SURFACTANT ENHANCED REACTION BETWEEN‬‬

‫‪BENZOCAINE AND P-‬‬
‫‪DIMETHYLAMINOBENZALDEHYDE: KINETIC STUDY‬‬
‫‪AND ITS ANALYTICAL APPLICATION‬‬

‫‪Rabah A. Khalil* and Sana A. Hussain‬‬

‫‪Department of Chemistry, College of Science, University of Mosul, Mosul, Iraq‬‬

‫اﻟﺨﻼﺻـﺔ‪:‬‬
‫ﻣﻦ اﻟﻤﻌﺮوف ﺑﺼﻮرة ﺟﻴﺪة ﺑﺄن ﻋﻘﺎر اﻟﺒﻨﺰوآﺎﺋﻴﻦ ﻻﻳﺬوب ﻓﻲ اﻟﻤﺎء‪ ،‬ﻟﺬﻟﻚ ﻓﺈن ﻣﻌﺎﻟﺠﺎﺗﻪ اﻟﻔﻴﺰﻳﺎﺋﻴﺔ واﻟﺘﻘﺪﻳﺮﻳﺔ ﺑﻮﺟﻮد اﻟﻌﺎﻣﻞ اﻟﻔﻌﺎل ﺳﻄﺤﻴًﺎ ﺳﻮف‬
‫ﺗﻌﻄﻲ ﻓﺮﺻﺔ ﺣﻮل إﻣﻜﺎﻧﻴﺔ اﺳﺘﺒﺪال اﻟﻤﺬﻳﺒﺎت اﻟﻌﻀﻮﻳﺔ اﻟﻤﻠﻮﺛﺔ ﻟﻠﺒﻴﺌﺔ ﺑﺎﻟﻤﺤﺎﻟﻴﻞ اﻟﻤﺎﺋﻴﺔ ﻟﻠﻤﻮاد اﻟﻔﻌﺎﻟﺔ ﺳﻄﺤﻴًﺎ‪ .‬وﻗﺪ ﺗﻢ ‪ -‬ﻓﻲ هﺬا اﻟﺒﺤﺚ ‪ -‬دراﺳﺔ ﺗﺄﺛﻴﺮ اﻟﻌﺎﻣﻞ‬
‫اﻟﻔﻌﺎل ﺳﻄﺤﻴًﺎ ﻣﻦ اﻟﻨﻮع اﻟﺴﺎﻟﺐ )ﺻﻮدﻳﻮم دودﺳﻴﻞ ﺳﻠﻔﻴﺖ‪ (SDS ،‬ﻋﻠﻰ ﺣﺮآﻴﺔ اﻟﺘﻔﺎﻋﻞ اﻟﺘﻜﺎﺛﻔﻲ ﺑﻴﻦ اﻟﺒﻨﺰوآﺎﺋﻴﻦ ﻣﻊ ﺑﺎرا‪-‬ﺛﻨﺎﺋﻲ ﻣﺜﻴﻞ اﻣﻴﻨﻮﺑﻨﺰﻟﺪﻳﻬﺎﻳﺪ‬
‫)‪ (DAB‬ﻏﻴﺮ اﻟﺬاﺋﺐ اﻳﻀﺎ ﻓﻲ اﻟﻤﺎء‪ .‬وﻟﻮﺣﻆ ﺑﺄن وﺟﻮد )‪ (5x10-3M‬ﻣﻦ اﻟـ‪ SDS‬ﻳﺰﻳﺪ ﺳﺮﻋﺔ اﻟﺘﻔﺎﻋﻞ ﻋﺸﺮﻳﻦ ﻣﺮة ﻋﻦ ذﻟﻚ ﻓﻲ ‪ %20‬اﻳﺜﺎﻧﻮل‪-‬ﻣﺎء‪ .‬وﺗﻢ‬
‫ﻣﻼﺣﻈﺔ زﻳﺎدة آﺒﻴﺮة ﺟﺪًا ﻓﻲ ﺣﺰﻣﺔ اﻻﻣﺘﺼﺎص ﻓﻲ اﻟﻤﻨﻄﻘﺔ اﻟﻤﺮﺋﻴﺔ ﻣﻊ إزاﺣﺔ ﺣﻤﺮاء ﺑﺴﺒﺐ وﺟﻮد اﻟﻌﺎﻣﻞ اﻟﻔﻌﺎل ﺳﻄﺤﻴﺎ )‪ .(SDS‬وأﻇﻬﺮت اﻟﻨﺘﺎﺋﺞ أن‬
‫هﻨﺎﻟﻚ ﻣﺤﺎﺳﻦ آﺒﻴﺮة ﻓﻲ اﻟﺘﻄﺒﻴﻘﺎت اﻟﺘﺤﻠﻴﻠﻴﺔ ﻧﺘﻴﺠﺔ ﻟﺘﻜﻮُن ﻣﻌﻘﺪ اﻧﺘﻘﺎل اﻟﺸﺤﻨﺔ ﺑﺴﺒﺐ وﺟﻮد اﻟـ‪ .SDS‬إن ﻇﺎهﺮة ﺗﻜﻮُن ﻣﻌﻘﺪ اﻧﺘﻘﺎل اﻟﺸﺤﻨﺔ ﺗﻤﺖ اﻹﺷﺎرة إﻟﻴﻬﺎ‬
‫وإﺛﺒﺎﺗﻬﺎ ﻓﻲ هﺬﻩ اﻟﺪراﺳﺔ ووﺟﺪ ﺑﺄﻧﻬﺎ ﺗﻌﺘﻤﺪ ﺑﺼﻮرة آﻠﻴَﺔ ﻋﻠﻰ ﺷﺤﻨﺔ ﺳﻄﺢ اﻟﻤﺬﻳﻞ‪ .‬وﺗﻢ ﺗﻄﻮﻳﺮ ﻃﺮﻳﻘﺔ ﺑﺴﻴﻄﺔ‪ ،‬ﺣﺴﺎﺳﺔ ودﻗﻴﻘﺔ ﻟﻠﺘﻘﺪﻳﺮ اﻟﻤﺎﻳﻜﺮوي ﻟﻠﺒﻨﺰوآﺎﺋﻴﻦ‬
‫ﻓﻲ ﻣﺤﻠﻮﻟﻪ اﻟﻤﺎﺋﻲ وﺑﻮﺟﻮد اﻟـ‪ .SDS‬ووﺟﺪ ﺑﺄن هﺬﻩ اﻟﻄﺮﻳﻘﺔ ﻳﻤﻜﻦ ﺗﻄﺒﻴﻘﻬﺎ ﻋﻠﻰ اﻟﺒﻨﺰوآﺎﺋﻴﻦ ﻓﻲ ﺣﺎﻟﺘﻪ اﻟﻨﻘﻴﺔ وﻓﻲ اﻟﻤﺴﺘﺤﻀﺮات اﻟﺼﻴﺪﻻﻧﻴﺔ ﺑﺪﻗﺔ وﺣﺴﺎﺳﻴﺔ‬
‫أﻋﻠﻰ ﻣﻦ ذﻟﻚ ﻓﻲ ﻃﺮﻳﻘﺔ دﺳﺘﻮر اﻷدوﻳﺔ اﻟﺒﺮﻳﻄﺎﻧﻲ )‪ .(BP‬وﻗﺪ اﺳﺘُﻨﺘﺞ ﻓﻲ هﺬا اﻟﺒﺤﺚ ﺑﺄن اﺳﺘﺒﺪال اﻟﻤﺬﻳﺒﺎت اﻟﻌﻀﻮﻳﺔ ﺑﺎﻟﻌﻮاﻣﻞ اﻟﻔﻌﺎﻟﺔ ﺳﻄﺤﻴﺎ ﻻ ﺗﻘﺘﺼﺮ‬
‫ﻓﺎﺋﺪﺗﻬﺎ ﻓﻘﻂ ﻋﻠﻰ اﻟﻨﻮاﺣﻲ اﻟﺼﺤﻴﺔ واﻟﺒﻴﺌﻴﺔ ﺑﻞ ﻟﻪ ﻣﺤﺎﺳﻦ أﺧﺮى ‪ ،‬ﻋﻠﻤًﺎ ﺑﺄن ﻣﺎدة اﻟـ‪ SDS‬اﻟﻤﺴﺘﺨﺪﻣﺔ ﻓﻲ هﺬﻩ اﻟﺪراﺳﺔ ﺗﻌﺘﺒﺮ آﺼﺪﻳﻖ ﻟﻠﺒﻴﺌﺔ ﻟﻜﻮﻧﻬﺎ اﻟﻌﺎﻣﻞ‬
‫اﻟﻔﻌﺎل ﺳﻄﺤﻴﺎ اﻟﻤﺴﺘﺨﺪم ﻓﻲ ﻣﻌﺎﺟﻴﻦ اﻷﺳﻨﺎن‪.‬‬

‫_______________‬
‫‪*Corresponding Author:‬‬
‫‪E-mail: rakhalil64@yahoo.com‬‬
‫_____________________________________________________________________________________________‬
‫‪Paper Received July 7, 2009; Paper Revised November 15, 2009; Paper Accepted December 9, 2009‬‬

‫‪July 2010‬‬ ‫‪The Arabian Journal for Science and Engineering, Volume 35, Number 2A‬‬ ‫‪55‬‬
 Rabah A. Khalil and Sana A. Hussain

ABSTRACT
Benzocaine is well known as an insoluble substrate in water. Therefore, its physical and analytical treatments in
the presence of surfactant may allow organic solvents to be replaced by aqueous surfactants solutions. The effect of
the presence of anionic surfactant (sodium dodecyl sulfate, SDS) upon the condensation reaction of benzocaine with
water insoluble p-dimethylaminobenzaldehyde (DAB) in aqueous solution has been investigated extensively. The
presence of 5x10-3M SDS increases the reaction rate by 20 times in contrast to that of a solution in 20% ethanol. A
substantial increase in absorption band due to the formed Schiff base followed by a red shift has been observed. A
great advantage in analytical application resulting from the presence of surfactant due to the formation of a charge
transfer complex has been reported. The latter phenomenon has been realized and proved in this study, which
depends merely on the charge of micelle surface. A simple, accurate, and sensitive method for microdetermination of
benzocaine in aqueous surfactant solution has been introduced. The proposed procedure for analysis of the presented
drug in its bulk form and pharmaceutical formulations is more sensitive and accurate than that of the British
pharmacopoeia method. It has been concluded that the replacing of organic co-solvents by surfactants was not only
captured for avoiding their pollution and toxic implication but there are some advantages gained from the presence
of surfactant molecules. The used surfactant compound (SDS) in this study is quite green and is commonly used as a
detergent in toothpaste.
Key words: benzocaine, surfactants, sodium dodecyl sulfate, green chemistry, solvents, micellar catalysis, kinetic,
analytical procedure, p-dimethylaminobenzaldehyde

56 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010
 Rabah A. Khalil and Sana A. Hussain

SURFACTANT ENHANCED REACTION BETWEEN BENZOCAINE AND P-

DIMETHYLAMINOBENZALDEHYDE: KINETIC STUDY AND ITS ANALYTICAL
APPLICATION

1. INTRODUCTION
Benzocaine (I) [ethyl 4-aminobenzoate (94-09-7)] is medically known as local and epidural anesthetic, which is
used to relieve pain associated with ulcers. Recently, we have investigated the effect of the presence of surfactant on
the kinetics of the condensation reaction between p-dimethyl amino benzaldehyde (DAB) with six sulfa drugs [1],
bromhexine drug [2], and three β-lactam antibiotics [3]. It was found that the presence of anionic surfactant (sodium
dodecyl sulfate, SDS) in these reactions produces two main advantages. First, a substantial enhancement was
observed in the rate of these condensation reactions. Second, there is a remarkable increase in the extinction
coefficient of the products in the visible region, which makes the reaction quite applicable in analytical
determinations [1,3,4]. A characteristic feature of benzocaine in contrast to the earlier studied drugs is its insolubility
in water. It should be noted that the reagent DAB is also insoluble in water in a similar manner to benzocaine.
Therefore, it seems interesting to investigate the effect of the presence of surfactant upon the reaction between
benzocaine and DAB, which are both merely water insoluble. From an analytical point of view, there are various
methods that were employed for determination of benzocaine. These procedures include potentiometric [5],
spectrophotometric [6], colorimetric [7], HPLC [8], and conductometric titration techniques [9].

H2N C O CH2CH3

(I)

In the present work, an attempt was made to achieve a simple, accurate, and sensitive method for determining
benzocaine in its bulk form and pharmaceutical preparations in the absence of organic solvents. In other words, the
above kinetic and analytical treatments of two insoluble substances in aqueous surfactant will certainly give an
environmental hope for eliminating the use of organic solvents. This is due to the fact that the latter solvents have
potential pollution and toxic implications. The possible great improvement gained through replacing the organic
solvents by the presence of surfactant in aqueous media is similar to that of ionic liquids [10,11]. However,
treatments with the ionic liquids possess more difficulty in applications based on economical and technical aspects in
contrast with that of aqueous surfactant solutions.
2. EXPERIMENTAL METHODS
Benzocaine was obtained in highly pure form from State Drug Industry (SDI), Samarra-Iraq. All other reagents
were analytical grade commercial products purified when necessary by standard procedures. Distilled water was
used for preparation of all solutions. All spectra for kinetic and analytical measurements for the reaction between
benzocaine and DAB were performed on UV-Visible Spectrophotometer Centra 5 (GBC Scientific Equipment)
equipped with thermostated cell holder. To control the temperature within ±0.1 oC, water thermostated Hakke NK22
was used. The stoke solutions were freshly prepared 0.2 and 0.02M solutions of benzocaine in ethanol (99.99%) and
0.05M SDS, respectively. Solutions of 0.1 and 0.02M DAB were also prepared in ethanol (99.99%) and in 0.05M
SDS, respectively.
Kinetic measurements for the condensation reaction were normally used solutions of 1.5 x 10-4M benzocaine in
20% ethanol/H2O (in the absence of SDS), and 5 x 10-5M benzocaine in the presence of SDS; with varying
concentrations of DAB. The former solution contained 0.087M KCl for buffering composition (with desired amount
of HCl) of the solution and to avoid the effect of ionic strength. Addition of KCl was not needed in the presence of
SDS due to its negative effect on SDS and the latter surfactant could serve the same purpose for buffering
composition [2]. Details of the experimental procedure were briefly illustrated in Reference 1. The observed rate
constants were calculated with integral equations from the experimentally obtained absorbance vs time profile. The
calculations were performed using Microsoft Excel program for linear regression analysis. Multiple linear regression
analysis was employed for determining the binding constants using Minitab 11 software.

July 2010 The Arabian Journal for Science and Engineering, Volume 35, Number 2A 57
 Rabah A. Khalil and Sana A. Hussain

3. RESULTS AND DISCUSSIONS

3.1. Effect of Surfactant on Reaction Kinetics
In general, the condensation reaction of amines with DAB to produce Schiff base is considered to be reversible
[2,3,12,13]. However, we have found that the latter phenomenon depends significantly upon the chemical structure
of amine [14]. In other words, the resonance effect plays a major role in the phenomenon of reversibility. For
example, the presence of two bromine groups in ortho- and para- positions of the bromhexine amino group leads to a
remarkable opposite reaction [2]. For sulphonamides, the reaction is somewhat completely forward [1], while the
non-aromatic amino group of β-lactam antibiotics also leads to a reversible reaction [3]. The chemical structure of
benzocaine is relatively similar to that of sulphonamides and, therefore, a mostly forward reaction was expected.
The formed Schiff base from the reaction of benzocaine with DAB in the presence of HCl gives an absorbance
maximum at 460nm. Kinetic study of this reaction was performed following the increase in absorption intensity of
the latter visible peak with time. No maximum was observed when one follows the relation between absorption vs
time. This actually indicates that the presented reaction is mostly forward. Such observation was confirmed through
the application of our previous probe for detecting the reversibility of the reaction [2,3]. The non-linear relation was
achieved through the application of the latter probe between ([benzocaine]/Aeq] vs (1/[DAB]) of correlation
coefficient (r) equal to 0.53, indicating the reaction is mostly forward; where Aeq is the absorbance of the Schiff base
at infinite time, and [benzocaine] and [DAB] are the concentrations of the reactants in 20% EthOH/H2O at pH 3. The
above relation was also applied in the presence of SDS at the same pH, which gives a linear line with (r) value equal
to 0.998. However, the value of the obtained equilibrium constant is equal to 12335.5M-1 and indicates clearly that
the reaction is mostly forward [15].
The order of the reaction was also investigated using both the integrated isolation method and the initial rates
differential procedure. The latter was based on the following equation:
log Rate = Intercept + n log C (1)

where n is the order of the reaction and C is the reactant concentration. Thus, estimating the rate of the reaction from
the initial slope of the relation between one of the reactants or products vs time at different C, one could calculate n
from the slope of the above relation.
Table 1 illustrates the results of determining the order of the presented reaction including the application of the
isolation method using usual integrated rate equations. The results show that the r2 and S.E. of first order are better
than that of second order for equal concentrations of both reactants. On the other hand, this was confirmed by the
isolation method (Table 1), which indicates clearly that the reaction is first order with respect to DAB and zero order
with respect to benzocaine. However, it is apparent from the results of Table 1 that the relations of all used orders are
linear. In other words, there are no substantial differences between the values of r2. Therefore, in order to evaluate
the order of this reaction in a more confident way, an application of differential method based on initial rate was
applied. The results (Table 2) of the latter method indicate that the reaction is first order, which is quite parallel to
the above investigations using the integrated rate equation method. In other words, the reaction is zero order with
respect to benzocaine and first order with respect to DAB (rate=kr[Bezocaine]o[DAB]). A similar order of reaction
was previously found for sulphonamides [1] in contrast to those of bromhexine [2] and β-lactams [3], which is the
steric effect in their structures that plays a major role in such observations.

Table 1. Observed Rate Constants (kr), Order of Reaction (n), Square Correlation Coefficient (r2), and
Standard Error (S.E.) of the Reaction Between Equal Reactants Concentrations With Application of Isolation
Method in the Absence of SDS at pH 3 and 20oC
[Benzocaine] [DAB] Order
kr Intercept r2 S.E.
(M) (M) (n)
1.0x10-4 1.0x10-4 1st 0.1132 s-1 -3.60 0.999 9.6x10-3
1.0x10-4 1.0x10-4 2nd 8.0843 M-1s-1 27.69 0.974 4.32
-4 -1
1.0x10 0.01 zero 0.1218 Ms 1.192 0.992 0.012
-4 -1
1.0x10 0.01 1st 0.2623 s -0.025 0.979 0.044
-4 -1
0.01 1.0x10 zero 0.0565 Ms 0.681 0.964 2.0x10-4
0.01 1.0x10-4 1st 0.1169 s-1 -0.047 0.998 0.069

58 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010
 Rabah A. Khalil and Sana A. Hussain

Table 2. Order of the Reaction of Benzocaine and DAB from the Application of Initial Rate Differential
Method in the Absence of SDS at pH 3 and 20oC
[Benzocaine] [DAB] Order
Intercept r2 S.E.
(M) (M) (Slope)
1.0x10-4 Various Conc. 1.004 1.0304 0.991 0.0587
Various Conc. 0.01 0.230 0.0709 0.463 0.0613
0.01 Various Conc. 0.984 2.9557 0.981 0.04212

The order of the reaction also was investigated in the presence of SDS using integrated rate equations as
illustrated in Table 3. The latter shows there is a consistency with that in the absence of SDS (in 20% EthOH), but
the results in the presence of SDS is more obvious than those of Table 1.
Table 3. Observed Rate Constants (kr), Order of Reaction (n), Square Correlation Coefficient (r2), and
Standard Error (S.E.) of the Reaction Between Equal Reactants Concentrations With Application of Isolation
Method in the Presence of 4.0x10-3 M SDS at pH 3 and 20oC
[Benzocaine] [DAB] Order
kr Intercept r2 S.E.
(M) (M) (n)
5.0x10-5 5.0x10-5 1st 0.6630 s-1 -1.549 0.992 0.067
-5 -5 -1 -1
5.0x10 5.0x10 2nd 31.992 M s 30.208 0.924 15.769
-6 -4 -1
5.0x10 5.0x10 zero 0.0208 Ms 0.294 0.992 1.8x10-3
5.0x10-6 5.0x10-4 1st 0.6170 s-1 -2.00 0.924 0.105
-4 -6 -3 -1
5.0x10 5.0x10 zero 4.8x10 Ms 0.264 0.962 1.0x10-3
5.0x10-4 5.0x10-6 1st 0.6780 s-1 -3.479 0.991 0.064

Equation 2 may represent the proposed mechanism of the presented reaction, while Equation 3 expresses the
measured or observed rate constant.

k1 k2
Benzocaine + Protonated DAB [Intermediate] Schiff base + H2O (2)
k −1 k −2
k 1k 2
k (observed ) = (3)
k −1 k −2
Figures 1 and 2 exhibit the effect of pH upon this reaction in the absence and presence of SDS. No remarkable
differences in the shape of the relationship is observed due to the presence of micelle. There is only substantial
increase in the rate and absorbance caused by the presence of SDS. The decreases of the rate constant due to
increases in pH may simply be related to the decrease in the concentration of the catalyst HCl (Figure 1), while the
relation between infinite absorbance vs pH shows a maximum (Figure 2). The decrease of infinite absorbance with
increasing pH is due to the deprotonation of imine nitrogen in which the absorption band at 460 and 467nm in the
absence and presence of SDS, respectively, is due to the latter protonated imine atom, while the decrease in infinite
absorbance with decreasing pH may be attributed to the protonation of the carbonyl oxygen atom, which then
eliminates the conjugation of the formed Schiff base according to the following structure:

H3C
+
OH

H N C N C OCH2CH3
H
H3C
H

We think that the above structure is quite reliable because the presence of ethanol (in the absence of SDS) will
definitely enhance the resistance of Schiff base towards hydrolysis at low pH. However, in the presence of SDS, the
above structure will be stabilized by the negative surface charge of the micelle. It should be noted that the highest
infinite absorbances ( A ∞ ) according to Figure 2 are at pH 2.5 and 2 in the absence and presence of SDS,
respectively. However, due to the relatively fast reaction at those pH (Figure 1) levels, which may create some errors
in measurements, we have selected pH 3 for kinetic investigations.

July 2010 The Arabian Journal for Science and Engineering, Volume 35, Number 2A 59
 Rabah A. Khalil and Sana A. Hussain

2.5
in absence of SDS
2
in presence of 4e-3M SDS

1.5

kr(1/s)
1

0.5

0
1.8 2.3 2.8 3.3 3.8
pH
Figure 1: Effect of pH on observed rate constants of 1x10-4 M benzocaine and DAB in absence of SDS;
and for the reaction of 5x10-5 M benzocaine and DAB in the presence of 4x10-3M SDS at 20oC

1.6
in absence of SDS
1.4
in presence of 4e-3M
1.2

1
A(inf)

0.8

0.6

0.4

0.2

0
1 1.5 2 pH
2.5 3 3.5 4

Figure 2: Effect of pH on infinite absorbances ( A ∞ ) of 1x10-4 M benzocaine and DAB in the absence of SDS;
and for the reaction of 5x10-5 M benzocaine and DAB in presence of 4x10-3M SDS at 20oC
The effect of temperature upon the presented reaction was also investigated and summarized in Table 4 in the
absence of SDS. The activation energy (E≠) and frequency factor (A) have been estimated in the absence of SDS,
which give values 31.53 k.J.mol-1 and 4.8x104 s-1 (r2=0.92; S.E.= 0.1385), respectively, while other related
thermodynamic functions of activation give values of 77.78 kJ.mol-1, 29.05 kJ.mol-1, -163.43 J.mol-1.K-1, and
2.35X10-14 M-1 for ∆G≠, ∆H≠, ∆S≠, and K≠ , respectively, at 20oC and pH 3.
Table 4. Observed Rate Constant of the Reaction of 1x10-4 M Benzocaine and DAB
at Different Temperatures and pH 3
T(oC) Kr(s-1) Intercept r2 S.E.
5 0.04995845 -2.912629 0.997 0.0064
10 0.07681273 -3.043235 0.997 0.0105
15 0.10010 -3.340305 0.997 0.0125
20 0.13563077 -3.441797 0.993 0.0290
25 0.14608830 -3.598769 0.994 0.0259
30 0.15260650 -3.743812 0.997 0.0166
Table 5 illustrates the effect of temperature upon the observed rate constant at different concentrations of SDS,
while Figure 3 explains the effect of the presence of SDS at different concentrations and temperatures. Clear maxima
are shown, which is attributed to the effect of concentration and dilution of bimolecular reactants by micelles. The
reaction also followed the Arrhenius equation in the presence of SDS, according to the results summarized in Table
6. The activation energy, frequency factor, and other related thermodynamic functions of activation in the presence

60 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010
 Rabah A. Khalil and Sana A. Hussain

of different amounts of SDS are reported in Table 7. The results show that there is a successive decrease in the
values of activation energy and frequency factor with increasing SDS concentrations (Table 7). The reason for the
former may be attributed to the considerable stability that is achieved from the positively charged hemiaminal
activation complex with the negative charge of surfactant and micelle surface, while the reason for the latter
(frequency factor) may be attributed to the hydrophobic interaction between each of benzocaine and DAB with the
non-polar part of SDS. Such interactions could definitely reduce the collisions between reactant molecules and,
therefore, reduce the values of A and ∆S≠. The relatively constant value of ∆G≠ with change of [SDS] may be related
to the change in SDS concentration and does not change the structure of the activated complex, while in the absence
of SDS (as mentioned above) the value of ∆G≠ is relatively higher than those of Table 7, which may be attributed to
the stability of the positively charged activation complex with the SDS negative charge even at concentrations less
than the CMC (CMC of SDS at pH 3 = 0.004M) [16]. On the other hand, there is a considerable increase in E≠ and A
values particularly at a relatively low concentration of SDS (Table 7) in contrast to those above in the presence of
ethanol. Such phenomenon was already observed for bromhexine [2], which may also be explained in terms of
concentration and dilution of reactant molecules. The concentration of molecules usually increases the number of
collisions (A) between molecules. The activation energy may also increase due to increases in electrostatic repulsion
between the positively charged intermediate molecules in comparison with that in the absence of SDS, while the
increase of SDS concentration dilutes the concentration of reactant molecules and, therefore, decreases the values of
E≠ and frequency factor as clearly shown in Table 7.

3.5 T=5C
3 T=10C
2.5 T=15C
kr (1/s)

2
T=20C
1.5
T=25C
1
0.5
0
0 0.01 0.02 0.03
[SDS]
Figure 3: Effect of SDS concentration upon the observed rate constant of the reaction of 5x10-5 M benzocaine
and DAB at different temperatures and pH 3

Table 5. Observed Rate Constant of the Reaction of 5x10-5 M Benzocaine and DAB
in the Presence of Different Amounts of SDS at Different Temperatures and pH 3

[SDS]
(M) 0.001 0.002 0.003 0.004 0.005 0.007 0.009 0.020

T(oC) kr (s-1)
5 0.3565 ---- 0.8464 0.9646 1.1466 1.0386 1.0953 0.6296
10 0.6798 0.7590 1.1434 1.4865 2.0333 1.5294 1.4999 0.6496
15 0.8147 0.8417 1.2167 1.8775 2.3598 1.7666 1.6797 0.6900
20 0.9098 1.2899 1.7544 2.4420 2.5917 1.9114 1.9413 0.8182
25 1.6910 1.9491 2.3672 2.5734 2.9578 2.4000 2.0600 ---

July 2010 The Arabian Journal for Science and Engineering, Volume 35, Number 2A 61
 Rabah A. Khalil and Sana A. Hussain

Table 6. Statistical Parameters Resulting from the Application of Arrhenius Equation to the Reaction of
5x10-5 M Benzocaine and DAB in the Presence of Different Amounts of SDS at pH 3
[SDS](M) Slope Intercept r2 S.E.
0.001 -5674.33 19.45 0.923 0.179
0.002 -5402.78 18.70 0.947 0.12
0.003 -4141.7 14.68 0.966 0.084
0.004 -4098 14.78 0.936 0.117
0.005 -3563.16 13.11 0.843 0.168
0.007 -3163.16 11.48 0.932 0.0928
0.009 -2536.66 9.28 0.922 0.08
0.02 -1413.5 4.58 0.873 0.05

Table 7. Activation Energy, Frequency Factor and Thermodynamic Functions of Activation for the Reaction
of 5x10-5 M Benzocaine and DAB in the Presence of Different Amounts of SDS at 25oC and pH 3

[SDS] E≠ A ∆G≠ ∆H≠ ∆S≠ K≠

M KJ/mol l/mol.s KJ/mol KJ/mol J/mol.K M-1

0.001 47.17 2.8x108 71.71 44.69 -90.62 2.73x10-13

0.002 44.91 1.3x108 71.37 42.43 -97.06 3.13x10-13
6
0.003 34.43 2.3x10 70.88 31.95 -130.56 3.82x10-13
0.004 34.0 2.6x106 70.67 31.52 -131.3 4.1x10-13
0.005 29.62 4.9x105 70.33 27.14 -144.85 4.7x10-13
4
0.007 26.29 9.6x10 70.85 23.81 -157.7 3.86x10-13
0.009 21.08 10721.4 71.22 18.6 -176.48 3.3x10-13

Application of the kinetic data listed in Table 7 for determining the binding constant between each of Benzocaine
and DAB with micelle has been carried out using the available developed methods [2,3,17]. Indeed, the values of
binding constants cannot be determined, which is mathematically impossible. In other words, the equation for
determining the binding constants for this reaction is not valid for water insoluble reactants. The phenomenon
suggests the need for doing extensive studies in order to obtain a reliable reason for the above invalidity (future
work).
Figure 4 shows the UV-vis spectrum of the Schiff base produced from the reaction of 5x10-5 M benzocaine and
DAB in the absence and presence of SDS. It is apparent that there is a substantial increase in the intensity of the
visible absorption band belonging to the protonated imine nitrogen due to the presence of SDS. The reason for such
an interesting phenomenon may be attributed entirely to the formation of charge transfer complex due to the
presence of SDS. This could be explained by the fact that the ionization potential of the surfactant negative group
( −SO 3− ) is far less than that of chloride ion (due to the presence of HCl; pH=3). Hence, there is great probability for
a negative charge transfer from −SO 3− group of micelle to the positively charged imine nitrogen, which is
responsible for such an absorption band [14].

62 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010
 Rabah A. Khalil and Sana A. Hussain

Figure 4: Shows the UV-vis spectrum of the Schiff base produced from the reaction of 5x10-5 M benzocaine and DAB at 20oC
and pH 3 (a) in the absence of SDS; 20% EthOH (b) in the presence of 0.004M SDS
3.2. Analytical Application
The above investigations suggest an analytical method for the determination of benzocaine as water insoluble
compound in aqueous media. The method was based upon the reaction of the drug with an excess amount of DAB in
acidic medium and the presence of SDS. The produced yellow color can be followed photometrically at an
absorbance maximum of 467nm. It is apparent from the above investigations that the molar ratio of benzocaine to
DAB is 1:1. Figure 2 already indicates clearly the optimum pH for analytical determinations, which is equal to 2.
Figure 5 illustrates the effect of changing SDS concentration upon absorbance. This indicates that the ideal
concentration of SDS for this purpose is 0.004M.

0.2

0.15
Abs

0.1

0.05

0
0 0.002 0.004 0.006 0.008 0.01
[SDS](M)
Figure 5: The relation between absorbance against SDS concentration for the reaction of 1x10-5M benzocaine
and 5x10-5M DAB at pH 2 and room temperature
The effect of DAB concentration upon the absorbance was also investigated and illustrated in Figure 6. Thus,
1x10-3M of DAB should be present in the reaction mixture to give the highest absorbance and to ensure the
quantitative determination of the drug at highest concentrations encountered in the calibrations. It should be noted
that we cannot exceed the latter concentration of DAB due to its limited solubility in 0.004M SDS. Figure 7 shows
the absorption spectra of the produced Schiff base against each of blank and distilled water together with the blank
against distilled water. Indeed, this could give significant support for the presented procedure. The order of addition
was studied and the results indicate that the order of (DAB/SDS+Benzocaine/SDS+HCl) gives the highest
absorbance.
The effect of the presence of other types of surfactants (in addition to SDS) on the absorbance was investigated
and illustrated in Table 8. Interestingly, the results give direct evidence for our proposal of the formation of a charge
transfer complex. This is due to the relative decrease in the absorbance by the addition of cationic (cetyl pyridinium
chloride mono hydrate, CPC) and nonionic (triton X-100) surfactants, while the anionic (sodium dodecyl benzene
sulphate, SDBS) surfactant shows a similar effect in comparison to that of SDS (Figure 5). Thus, the last significant
advantage of using surfactant solution in analytical determinations is also quite parallel to green chemistry through
avoiding the use of organic solvents.

July 2010 The Arabian Journal for Science and Engineering, Volume 35, Number 2A 63
 Rabah A. Khalil and Sana A. Hussain

0.6

0.5

0.4

Abs
0.3

0.2

0.1

0
0 20 40 60 80 100 120
[DAB]xE-5

Figure 6: Effect of DAB concentration on the absorbance for 1x10-5M benzocaine in presence
of 0.004M SDS at pH 2 and room temperature

Figure 7: Absorption spectra of 1x10-5M benzocaine in 0.001M/0.004M DAB/SDS at room temperature and pH = 2, (a) against
blank, (b) against distilled water, (c) blank against distilled water
Table 8. Effect of Addition of Other Type of Surfactants Including Anionic (SDBS), Cationic (CPC), and
Nonionic (Triton X-100) on the Absorbance of 2x10-5M Benzocaine in 0.001M/0.004M DAB/SDS at Room
Temperature and pH2
ml of surfactant solution added/absorbance
[Surfactant solution]
0 1 2 3
SDBS 0.001M 0.8742 0.8946 0.8771
CPC 0.001M 0.8168 0.8031 0.7739 0.6765
Triton X100 1% 0.8045 0.7282 0.6844

The stability and reproducibility of the mixture was studied at three different concentrations of benzocaine
(2.0x10-6, 5.0x10-6 and 2.5x10-5 M). The results indicate that the color obtained possesses a good stability and
reproducibility. The latter reproducibility have the values 7.75x10-4, 2.6x10-3, and 8.33x10-3 for the above three
concentrations, respectively. In addition, 20 minutes development time was selected as the optimum in the presented
procedure [15].
3.3. Analytical Procedure
1ml of 0.01M/0.02M of DAB/SDS reagent was pipetted into a 10ml volumetric flask. A known volume of
sample or stock benzocaine in 0.03M SDS was added and then followed by addition of a volume of 0.025 SDS that
made the final concentration of SDS in the 10ml equal to 0.004M. 1ml of 0.546M HCl was placed in the flask to
obtain the desired pH (pH=2). The mixture was diluted to 10ml and then shaken and left to stand at room
temperature for 20min. The absorbance was measured at 467nm against a reagent–blank solution.

64 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010
 Rabah A. Khalil and Sana A. Hussain

The typical calibration data for benzocaine obtained from linear regression analysis of absorbance vs
concentration of the presented procedure give Beer’s law limit value (5X10-7-3X10-5M or 0.0825-4.9558µg/ml),
slope (40763.34), intercept (0.015833), square of correlation coefficient (r2= >0.999 ), and standard error (0.00557)
at room temperature. The extinction coefficient (εsb), recovery%, and other analytical parameters for three different
concentrations of benzocaine in bulk form are listed in Table 9.
Table 9. Extinction Coefficients, Mean Recovery % and Other Analytical Parameters for Three Different
Concentrations of Benzocaine in Bulk Form Using the Presented Method at Room Temperature
Relative Sandell's
[Benzocaine] εsb Mean Standard Relative
standard Index
(M) (l/mole.cm) recovery%* deviation error %
deviation % (g/cm2)
2.0x10-6 48500 99.63 1x10-4 -0.369 ±0.103 0.00340
5.0x10-6 43600 99.248 1.11x10-4 -0.752 ±0.052 0.00378
2.5x10-5 41112 99.312 2x10-4 -0.688 ±0.0199 0.0040
*Mean of four determinations
The effect of the presence of some common pharmaceutical additives such as starch, glucose, lactose, dextrose,
gum acacia, glycerol, and vanillin in three different amounts (200, 400, and 800µg) has been explored. The results of
recovery% indicate there is no significance interferences produced by these foreign substances [15]. Applications of
the presented method for determination of benzocaine in two pharmaceutical preparations, including throat lozenges
and lozenges of benzocaine in three different concentrations, were carried out (Table 10).
Table 10. Application of the Presented Method for Determination of Three Different Concentrations of
Benzocaine in Some Pharmaceutical Formulations at Room Temperature
Product Conc. (M) Recovery %*
3x10-6 100.99
-6
Throat lozenges 6x10 99.495
-5
1.8x10 98.656
-6
3x10 101.648
Lozenges of -6
6x10 99.30
Benzocaine
-5
1.8x10 98.549
* Mean of four determinations
Indeed, it is apparent that the proposed analytical procedure possesses many advantages in contrast to the official
or standard method of British Pharmacopoeia [18]. The latter method, based on potentiometric titration with sodium
nitrate, gives a limit of detection equal to 50mg(5000µg)-over, recovery% 100.629, standard deviation 5.77x10-2,
relative error% +0.629, and relative standard deviation% equal to ± 0.789 . On the other hand, application of the
BP method to pharmaceutical preparations gives recovery% values for throat lozenges and lozenges of benzocaine
66.13 and 99.14, respectively. Thus, the proposed procedure is more sensitive than the official method, in addition to
the relative simplicity of the reagent and apparatus, in contrast to BP procedure.
4. CONCLUSIONS
In general, the results of this paper indicate the possibility of replacing organic solvents by aqueous surfactants
with considerable advantages in addition to that belonging to green chemistry. The latter branch of chemistry
recommends avoiding the use of organic solvents for environmental considerations. The present results suggest
replacing the organic co-solvents by aqueous surfactant solution in analytical applications; noting that the suitable
type of surfactant can be selected in such a way that it produces a charge transfer complex. Another advantage of
using surfactants in the last treatments, beyond sensitivity and accuracy, is that the organic co-solvents may cause a
leaching of plasticizer belonging to the automation analytical procedures. It was proved in the present study that the
substantial enhancement of absorption band of the product is due to the charge transfer complex. In order to give
recommendations about the use of aqueous surfactant instead of organic solvents for organic synthesis, we have to
do a next step concerning the isolation of the formed Schiff base. Recently, aqueous surfactant solutions were used
in the extraction of alkaloids from plant instead of organic solvents [19]. The results prove that the solution of SDS
is better and more efficient than organic solvents. Finally, the used surfactant compound (SDS) in this study is quite
green so it is non-volatile and commonly used as a detergent in toothpaste.

July 2010 The Arabian Journal for Science and Engineering, Volume 35, Number 2A 65
 Rabah A. Khalil and Sana A. Hussain

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66 The Arabian Journal for Science and Engineering, Volume 35, Number 2A July 2010

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