ANALYTICAL SCIENCES MAY 2003, VOL.

19 691
2003 © The Japan Society for Analytical Chemistry

Determination of Dipyrone in Pharmaceutical Products by

Flow Injection Analysis with Potentiometric Detection
Jackson S. ALBUQUERQUE,* Valdinete L. SILVA,*† Fernando LIMA,* Alberto N. ARAÚJO,** and
Maria Conceição B. S. M. MONTENEGRO**

*Department of Chemical Engineering/Environmental Engineering and Quality Laboratory-UFPE,

50740-521-Recife, Brazil
**CEQUP/Department of Physical Chemistry, Faculty of Pharmacy,
Rua Aníbal Cunha 164, 4050-Porto, Portugal

This work describes an FIA potentiometric procedure for the quantification of dipyrone in pharmaceutical products. For
the detector, a tubular electrode comprising a polymeric membrane containing tetraoctylammonium as an electroactive
material (5% w/w), dibutylphtalate as a mediator solvent (65% w/w) and PVC (30% w/w) directly applied above a
graphite conductor support was used. This unit was incorporated into a monochannel FI-system with a 0.1 mol/L
phosphate buffer solution (pH = 5.2) as the carrier solution. The electrode showed a linear response from 8.0 × 10–4 to
10–1 mol/L dipyrone, a slope of 62.1 ± 0.2 mV/dec in pH 5.2 units, an injection volume of 500 µL and a carrier flow-rate
of 6 mL/min. This procedure was applied to the analysis of pharmaceutical formulations (oral and injectable) containing
dipyrone; the obtained results gave a relative error of less than 3.9% and coefficients of variation less than 1% and 5%,
respectively, for the FIA and classical iodometric methods.

(Received May 21, 2002; Accepted December 12, 2002)

which can be applied to the on-line analysis of pharmaceutical

Introduction products.21,22 To the best of our knowledge, the only electrode
selective to dipyrone described in the literature to date23 is made
The widespread dosification and/or adulteration of up of a polymeric membrane, and containing a solution and
commercially available pharmaceutical preparations demand internal reference electrode. However, no analytical application
reliable methods for drug determination, that are preferably is mentioned concerning this unit, probably because it exhibits a
selective, rapid and can be undertaken with simple equipment. high electrical resistance (2 × 105 S), which contributes to a
This is the case of dipyrone[(2,3-dihydro)1,5-dimethyl-3-oxo-2- slow response time (about 1 min). This makes it inadequate for
phenyl-1H-pyrazol-4-yl)methylamino]methanesulfonate mono- discrete utilizations, especially in continuous flow. In an effort
hydrate, (Fig. 1), which is widely used in therapeutics as an to prevent these drawbacks, the construction and evaluation of a
analgesic, antipyretic and antispasmodic drug. Due to its tubular detector sensitive to dipyrone was proposed in this
dangerous human side effects, it is neither available in most work, and whose characteristics favor the development of a
Northern European markets nor in the U.S., although it methodology based on the FIA technique with potentiometric
constitutes a standard analgesic drug in many other parts of the detection for routine applications in the analysis of
world, namely Brazil. Hence, a large number of methods for its pharmaceutical formulations.
quantitative analysis have been proposed, including titrimetry in
aqueous1–3 and non-aqueous media,4–6 chromatography,5–10
luminescence,11,12 spectrophotometry,13–16 atomic-absorption Experimental
spectrometry,17 near-infrared spectrometry,18 amperometry and
polarography.19,20 Reagents
Potentiometry with ion-selective electrodes is now a well- All prepared solutions were made up of analytical-grade
established method that, when applied to the analysis of
pharmaceutical products, can be considered to be an
advantageous alternative to others normally used, owing to its
selectivity, reduced implementation costs and versatility of
application. Beyond this, the possibility of conferring a tubular
form to the detection units has permitted its utilization in
continuous-flow analysis systems (FIA), giving rise to simple,
easily operated manifolds involving economic equipment,
† To whom correspondence should be addressed.

E-mail: vlins@ufpe.br Fig. 1 Chemical structure of dipyrone.

692
Fig. 2 FI-system for dipyrone determination in pharmaceutical
products: C, carrier (sodium phosphate buffer, 0.1 mol/L pH 5.2); S,
sample (dipyrone); V, injection valve; P, peristaltic pump; GE,
grounding electrode; ESI, tubular electrode; RE, reference electrode;
D, detector; R, recorder; W, waste.
chemicals, and double-distilled water was used. Dipyrone
sodium salt, dibutylphthalate and tetraoctylammonium bromide
were obtained from Sigma Chemical Co. and pharmaceutical
samples were obtained from local drugstores.
Two different commercial 0.1 mol/L phosphate buffer
solutions of pH 6.7 and 5.2 (Merck) were used to prepare the
ionic sensor and in the flow system as a carrier solution,
respectively.
A stock solution containing 0.1 mol/L of dipyrone was
prepared by rigorous weighing of the solid and dilution in a
phosphate buffer solution. The less-concentrated calibration
solutions were prepared from this by rigorous dilution of the
same buffer solution.
The solutions from solid samples with a dipyrone
concentration near 5 × 10–3 mol/L were prepared by the
pulverization of 10 tablets of each formulation, weighing the
quantity of resulting powder and diluting in a phosphate buffer
solution of pH = 5.2. Injectable samples (2.0 mL = 500 mg of
dipyrone) were prepared by dilution of a rigorous product
volume in phosphate buffer in order to obtain a solution with an
amount equivalent to 5 × 10–3 mol/L dipyrone.
To analyze the samples by the classical idometric method, the
preparation procedure recommended by Brazilian
Pharmacopoeia24 was followed. For this propose, the samples
were prepared by weighing an equivalent of 0.2 g of the product
(tablet), or 2.0 mL of the injectable before diluting this with
water up to a final volume of 5.00 mL.
For an analysis by the conventional method, a 0.1mol/L
iodine solution, previously standardized (each mL of the
iodometric solution was equivalent to 17.5 mg of dipyrone, after
standardization with sulfur trioxide), was used.
Equipment
A schematic diagram of the flow-injection set-up is given in
Fig. 2, which comprises an Ismatec IPC-8 peristaltic pump to
impel the solutions, and a home-made proportional injector25 to
introduce samples into the flow system. A coiled PTFE tubular
reactor (100 cm long and 0.8 mm internal diameter), connected
the injection device to the detector. The flow-through electrode
supports and grounding electrode were also home made.26
Potentiometric measurements were performed with a Radelkis
208/1 pH meter (sensitivity of ± 0.1 mV) equipped with an SCE
reference electrode of the same brand Ref. OP-0830P. A single
pen Graphic 1000 Lloyd Instruments recorder was coupled to a
pH meter to record the obtained analytical signals.
Membrane preparation and electrode assembly
To prepare the ionic sensor, 0.5468 g of tetra-octylammonium
bromide dissolved in about 10.0 mL of acetone was added to
10.0 mL of 0.1 mol/L dipyrone in a 0.1 mol/L sodium
ANALYTICAL SCIENCES MAY 2003, VOL. 19
Fig. 3 Effect of the buffer pH (pH 4.5 to 7) on the potentiometric
response of the detector. The flow-rate was 4 mL/min and the
sample volume was 500 µL dipyrone.
phosphate buffer (pH 6.7) for 15 min in an inert atmosphere
(N2). The resulting precipitate, after the evaporation of acetone,
was filtered and washed with double-distilled water, and dried
in a dessicator at room temperature, protected from the light.
Approximately 0.020 g of the precipitate was dissolved in 0.26
g of dibutylphthalate. This solution was mixed with 0.12 g of
PVC previously dissolved in 6.0 mL of tetrahydrofuran. The
resulting membrane was applied directly to a conductive
support composed by a mixture of epoxy resin and graphite,
following a previously described methodology.27 After drying
the membrane, the electrode was inserted into the FIA system,
and the conditioning of the membrane was made with a 0.02
mol/L dipyrone solution flowing at a rate of 1 mL/min for about
1 h.
Results and Discussion
To evaluate the working characteristics of the constructed
tubular detectors, and to make the applicability of the developed
system viable for the determination of dipyrone in
pharmaceutical formulations, the FI manifold depicted in Fig. 2
was developed. Following this, the optimization of several
physicochemical and hydrodynamic parameters of the system
was conducted using an univariated method, namely the pH of
the carrier solution, the injection volume and the flow rate. The
influence of the pH of the carrier solution on the potentiometric
response of the electrodes was evaluated over the interval of 4.5
– 7.0 units by injecting different solutions of dipyrone in the
range of 1 × 10–3 to 10–1 mol/L into a carrier adjusted to a
convenient pH. As depicted in Fig. 3, a rapid increase was
observed in the slope of the obtained calibration lines, from
about 28 mV/dec to 60 mV/dec, when the solution pH increased
from 4.5 to 5.2, followed by an equally accentuated fall for
greater pH values. Under these conditions, the use of a carrier
buffer solution with the pH adjusted to 5.2 units was chosen.
The influence of the injection volume on the performance of the
detector response was assessed by proceeding to intercalation of
volumes between 100 and 1000 µL of the dipyrone calibration
solutions, fixing the flow rate at 4 mL/min and using as carrier a
0.1 mol/L phosphate buffer solution with the pH adjusted to 5.2.
An obtained progressive increase in the intensity of the
analytical signals was verified, but for volumes greater than 500
µL they remained constant due to a lower sample dispersion
during transport up to the detector. A study of the influence of
the flow rate was carried out over a range between 1 and 8
mL/min (Fig. 4) and for two dipyrone concentration levels. A
ANALYTICAL SCIENCES MAY 2003, VOL. 19 693

Table 1 Potentiometric selectivity coefficients (log Kpot) for a

tubular dipyrone electrode
Interferent 1 × 10–3 mol/L 1 × 10–2 mol/L

Lactose –0.41 ± 0.05 –1.14 ± 0.09

Sulfate –0.33 ± 0.08 –1.14 ± 0.13
Chloride 0.08 ± 0.21 –0.42 ± 0.08
Citric acid 0.59 ± 0.23 0.05 ± 0.11
Nitrate 1.16 ± 0.14 1.14 ± 0.24
Bisulfite –0.40 ± 0.18 –10.30 ± 0.22
Phosphate –0.40 ± 0.08 –1.3 ± 0.14

Table 2 Determination of dipyrone in pharmaceutical

Fig. 4 Effect of the flow rate of the stream. Sodium phosphate preparations using the proposed system
buffer (0.1 mol/L pH 5.2) was used the carrier solution and the
sample volume was 500 µL. Obtained value/mg
Commercial
product Iodometric Relative
Stated value FI-method
method error
Conmel 320 mg/tablet 315 ± 3 306 ± 16 2.9%
Novalgina 500 mg/tablet 490 ± 4 489 ± 4 0.2%
Dipirona
LAFEPE 500 mg/2 mL 478 ± 5 497 ± 5 –3.9%

The results compared with those obtained by the iodometric method.

presents high selectivity for the great majority of species

studied, except for nitrate ions. However, these are not present
in the formulations. For this reason, they do not prejudice the
quality of the determinations.

Fig. 5 Calibration curve for a dipyrone electrode obtained after
system optimization.
flow rate of 6 mL/min was chosen as being optimum, reflecting
a compromise between the analytical sensitivity and the
sampling rate. Under these conditions, the system permitted
analyses to be carried out at a rate of 108 samples/h. The
electrodes presented a practical detection limit equal to 9.0 ×
10–4 mol/L, a linear response limit of 8.0 × 10–4 mol/L and a
slope of 62.1 ± 0.2 mV/decade with an intercept of 200.42 ±
1.24 mV and a correlation coefficient of 0.9997 (Fig. 5). In
general, these values were similar to those presented by the
authors for a previously described conventional electrode.23
Nevertheless, it should be pointed out the very low response
time for the developed tubular electrode, compared with the
conventional one23 was in-turn reflected in the elevated
analytical rates that could be obtained (of about 250 samples/h).
The reproducibility of the units potential was also high, giving
rise to analytical signals throughout the working day that did not
differ by more than 0.3 mV/day.
Study of interferences
The influence of some organic and inorganic species, namely
nitrate, chloride, sulfate, phosphate, citrate and lactose on the
response of the detector was evaluated through the coefficients
of the potentiometric selectivity (log Kpot) by the separated
solutions method28 for concentrations of dipyrone and the
interferent equal to 1 × 10–3 mol/L and 1 × 10–2 mol/L. The
values indicated in Table 1 demonstrate that the electrode
Analytical applications
With the FIA system was implemented, different commercial
product samples was analyzed, including tablets and injectables.
Table 2 gives the obtained values, and compares them with
results provided from an analysis of the same products by the
classical idometric method.24 This was based on the addition of
5.0 mL of 0.02 mol/L chloridic acid to the samples, followed by
titration at a temperature lower than 20˚C, with a 0.1 mol/L
iodine solution. An analysis of the results concludes a good
agreement between the two methods (relative error lower than
3.9%) with the coefficients of variation being lower than 1%
and 5%, respectively, for the FIA and idometric methods.
Conclusions
The obtained results show that it is possible to use a
potentiometric unit of good working characteristics in a tubular
form that, when inserted into a simple flow system, makes
possible an automatic analysis of pharmaceutical formulations
containing dipyrone with increased accuracy and analytical
precision. The developed method is quickly carried out, simple
to operate, does not require specific sample treatments and can
be implemented in industrial pharmaceutical laboratories for
routine laboratorial analysis.
Acknowledgements
To the program for bilateral cooperation ICCTI/CAPES, CNPq,
LAFEPE and FACEPE.
694
References
1. J. Zeng, Yaoxue Xuebao, 1982, 17, 841.
2. K. Fukamachi, R. Nakagawa, M. Morimoto, and N.
Ishibashi, Bunseki Kagaku, 1975, 24, 428.
3. B. A. El-Zeany, M. M. Amer, A. M. Taha, and A. O. El-
Sawy, Analyst, 1982, 107, 1054.
4. S. S. M. Hassan, Arab. Gulf. J. Sci. Res., 1986, 4, 127.
5. M. K. Srivastava, V. P. Verma, and I. C. Shukla, Natl.
Acad. Sci. Lett., 1986, 9, 229.
6. B. Marciniec, J. Dorszeweska, and B. Wojciechowska,
Farm. Pol., 1990, 46, 11.
7. T. Aburjai, B. I. Amro, K. Aiedeh, M. Abuirjeie, and S. Al-
Khalil, Pharmazie, 2000, 55, 751.
8. D. Agbaba, O. Grozdanovic, L. Popovic, S. Vladimirov,
and D. Zivanov-Stakic, J. Planar Chromatogr. Mod. TLC,
1996, 9, 116.
9. H. L. Rau, A. R. Aroor, and P. Gundu-Rao, Indian Drugs,
1991, 29, 94.
10. G. Magyar-Pinter and Z. Elek-Voros, Acta Pharm. Hung.,
1990, 60, 77.
11. Y. Huang, C. Zhang, X. R. Zhang, and Z. J. Zhang, J.
Pharm. Biomed. Anal., 1999, 21, 817.
12. T. Perez-Ruiz, C. Martinez-Lozano, V. Tomas, and J.
Carpena, Microchem. J., 1993, 47, 296.
13. A. G. Tininis, A. Leandro, H. R. Pezza, C. B. Melios, and
L. Pezza, Anal. Lett., 2000, 33, 2901.
ANALYTICAL SCIENCES MAY 2003, VOL. 19
14. F. L. Zhao, B. Z. Xu, Z. Q. Zhang, and S. Y. Tong, J.
Pharm. Biomed. Anal., 1999, 21, 355.
15. E. Dinc and F. Onur, Anal. Chim. Acta, 1998, 359, 93.
16. K. K. Verma, A. Jain, and K. K. Stewart, Anal. Chim. Acta,
1992, 261, 261.
17. L. Lahuerta-Zamora and J. Martinez-Calatayud, Talanta,
1993, 40, 1067.
18. Y. L. Ren, Y. H. Gou, Z. Tang, P. Yiu, and Y. Guo, Anal.
Lett., 2000, 33, 69.
19. T. Perez-Ruiz, C. Martinez-losano, and V. Tomas, J.
Pharm. Biomed. Anal., 1994, 12, 1109.
20. F. Belal, Electroanalysis, 1992, 4, 589.
21. M. N. M. P. Alçada, J. F. C. Lima, and M. B. S. M.
Montenegro, Analyst, 1994, 119, 2327.
22. J. L. F. C. Lima, M. N. M. P. Alçada, and M. C. B. S. M.
Montenegro, Anal. Chim. Acta, 1993, 283, 657.
23. L. Cai, J. Pan, and S. Cui, Anal. Proc., 1987, 24, 339.
24. Brazilian Pharmacopea, 3rd ed., 1977, Organização Andrei
Editora S. A, São Paulo, Brazil, 408.
25. F. J. Krug, H. Bergamim, and E. A. G. Zagatto, Anal.
Chim. Acta, 1986, 179, 103.
26. S. Alegret, J. Alonso, J. Bartroli, A. A. S. C. Machado, J. L.
F. C. Lima, and J. M. Paulis, Quim. Anal., 1987, 6, 278.
27. J. L. F. C. Lima, M. C. B. S. M. Montenegro, and A. M. R.
Silva, J. Flow Injection Anal., 1990, 7, 19.
28. IUPAC, Analytical Chemistry Division Commission on
Analytical Nomenclature, Pure Appl. Chem., 1981, 53,
1907.