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7chnru. 28. 377to381. Vol. pp.

1981 Prtnted Great Britain in

Pcrgamon Press Ltd


Institute of Chemical and Pharmaceutical Research, 74351 SOS.Vitan 112, Bucharest, Romania
(Received 20 October 1980. Accepted 31 December 1980)

Summary-Liquid-membrane ion-selective electrodes made with the ion-association complexes of novocaine with tetraphenylborate or dipi~~mine are proposed for use in the determination of novocaine by direct potentiometry or by potentiometric titration with sodium tetraphenylborate. The titration method is preferred, and the mean relative error for 4-12 mg of novocaine hydrochloride is 0.5%.

The control of drug quality is a branch of analytical chemistry that has a wide impact on public health, so

the development of reliable, quick and accurate methods for determination of the active ingredient is welcomed. In recent years, ion-selective membrane electrodes have been used more and more in drug-quality control, but no pharmacopoeia has yet introduced their use for assays, though this will probably be done in the next few years. An important advantage of ion-selective membrane electrodes is that they can, in principle, be designed for any ionic species, even for organic ions with complicated structures and high molecular weight.- Novocaine (neocaine, procaine, ethocaine, syncaine, kerocaine, etc.-2diethylaminoethyl p-aminobenzoate hydr~hlorid~is a short-acting local anaesthetic. It is less toxic than cocaine, but because of its poor penetration of intact mucous membranes is useless for surface application. A rapid specific method applicable to the determination of novocaine in the presence of the basic or-

ganic or inorganic species often accompanying it in pharmaceutical preparations is needed because most of the available methods are susceptible to environmental reaction conditions and suffer from lack of specificity, sensitivity and stoichiometry. Most pharmacopoeias- recommend determination of the pure substance by diazotization of the primary Gino-coup by titration with O.lN sodium nitrite. Determination of the active ingredient in pharmaceutical preparations requires a preliminary extraction followed by ultraviolet spectrophotometry or densitometry, but the method is tedious and rather inaccurate. We describe here a simple, rapid and accurate method for the determination of novocaine in some pharmaceutical preparations, based on the use of a novocaine-selective membrane electrode. Construction of other nov~ine-~nsitive electrodes has been described in the literature.1.12 An Hg+ or Ag+selective membrane electrode has also been used for evaluating the purity of this drug.i3*i4 Our novocaine-selective membrane electrodes are based on liquid membranes consisting of the ion-association complexes I and Ii.
1 r -1



I2 J Lcc,~~ I4 B

Membrane for electrode A



12] bO,N -4

0 N4



Membrane for electrode B

* National Institute of Chemistry, 77208 SpI. Independentei 202, Bucharest, Romania. 377


D. NEGOIU.M.S.IONESCU V.V. C~~FRE-( and EXPERIMENTAL tration. Determine the unknown concentration from the calibration curve. Potentiometric titration. Place the electrodes in the sample solution (3&40 ml, concentration Cc. 10m3M, pH 5.0). then titrate with O.OlM sodium tetraphenylborate. The end-point corresponds to the maximum slope on the E cs. volume titration curve. (I ml of O.OlM tetraphenylborate is equivalent to 2.73 mg of novocaine hydrochloride).
Injectable aqueous solutions (Gerouital and Aslaoital).

Direct potentiometric measurements were made at room temperature in mechanically stirred solutions, with a digital pH/mV-meter (Pracitronic) and the novocaine-selective electrode and a saturated calomel electrode (SCE) (type K401, Radiometer). The SCE was connected to the sample solution by a saturated sodium nitrate bridge. The pH measurements were made with a G 202 C glass electrode (Radiometer) and an SCE. The potentiometric titration curves were recorded with an automatic outfit composed of a TTT2 titrator. ABU I2 autoburette and SBR2c titrigraph recorder (Radiometer). with an SCE and the two novoCaine-selective electrodes. An Ag+/S--crystal membrane electrode (Model 94-16, Orion) was used for argentimetric titration of the sodium tetraphenylborate solution (O.OlM) for standardization purposes.

All reagents used were of analytical-reagent grade. Sodium tetraphenylborate solution. 0.01M. Dissolve 0.85 g of sodium tetraphenylborate in the minimum of distilled water, filter, and dilute to 500 ml with distilled water. Standardize by potentiometric titration with O.OlM silver nitrate (with a silver-selective electrode and SCE).
Preparation of the liquid membranes

Transfer the contents of an ampoule with distilled water into a 25-ml standard flask and dilute to volume. Take 3.0 ml of this solution in a SO-ml beaker, add 20 ml of distilled water and adjust the pH to 5.0 with hydrochloric acid and/or sodium hydroxide solution. Titrate potentiometrically with O.OlM tetraphenylborate as above. The weight of novocaine hydrochloride in the ampoule is V x f x 25 x 273/3 mg where V is the volume of sodium tetraphenylborate solution used and S is the molarity of the titrant. Tablets (Gerocital H, and Aslacital). Weigh (W m ) and finely powder 20 tablets. Transfer about a quarter 7accurately weighed, w mg) to a IO@ml beaker..extract with 25 ml of distilled water. filter into a 200-ml standard flask and dilute to volume with distilled water. Take 5.0 ml from this solution in a 50-ml beaker, add 20 ml of distilled water. adjust to pH 5.0 and titrate potentiometrically as above. The average weight of novocaine hydrochloride per tablet = V x f x 546 x W/w mg.

Mix 10 ml of O.OlM novocaine hydrochloride solution with 20 ml of sodium tetraphenylborate solution. Filter off the precipitate on a porosity-4 sintered-glass crucible, wash with distilled water and dry at 100 for I hr. Dissolve the precipitate in 25 ml of nitrobenzene. Dry the solution by passage through filter paper and anhydrous sodium sulphate, then dilute to 100 ml with nitrobenzene. This is the liquid membrane (10e3M) for electrode A. (b) Nouocaine-dipicrylaminate. To obtain the liquid membrane for electrode B. mix IO ml of O.OlM notocaine hydrochloride in a separating funnel with IO ml of O.OlM dipicrylamine (dissolved in dilute sodium carbonate solution by heating). The novocainedipicrylaminate ionassociation complex is precipitated. Add 25 ml of nitrobenzene and shake the two phases for I5 min. Separate the phases and dry the organic layer by passage through filter paper and anhydrous sodium sulphate. Transfer to a 10%ml flask and dilute to volume with nitrobenzene. Nitrobenzene is a suitable solvent for liquid membranes, because of its high partition coefficient, and its high dielectric constant which gives high conductance to the membrane. Also. it is of low volatility, which is reflected in the maintainance of a constant concentration of the membrane and consequently of the conditional standard potentials,
(a) Nococaine-tetraphenylborate. EL. Constructiorl of the nooockine-selective membrane electrodes


The electrode function

The emf measurements were made with the electrochemical cell: novocaineselective $~c~~~~ (A or B) novocaine . HCl satd. (C) I = 0.1 (NaNO,) ; ~~l~~
(PH 5.0)

* SCE (1)

where C is the novocaine concentration, 10-l to 10m6M. The emf is given by:
E ICAl %(A,

ranging from

f 0.047 log[novocaine+] and

E I(B) + Km,

(for electrode A) (1)



(for electrode B) (2)

The construction of the electrodes has been described previously 5.6 and consists of impregnating the support material (a graphite rod I5 mm long and 6.5 mm in diameter. made water-repellent) with the electrode material dissolved in nitrobenzene. The internal reference electrode is eliminated by using a stainless steel wire introduced into the graphite rod. The electrodes are stored in the relevant liquid membrane solutions and are washed with distilled water between measurements,
Recommended Direct procedures

Prepare lo-, 10-j and 10e4A4 standards (I = 0.1, NaNOa, pH 5.0) by serial dilution of O.lM novocaine hydrochloride. Place the novocaine and reference electrodes in the stirred standard solutions in the order IO--. 10m4, IO-M. Plot E against log concenpotentiometry.

where the Eb values are the conditional standard potentials for electrodes A and B, under the conditions of use of cell (I). The measurements were carried out at constant ionic strength (I = 0.1, NaNO,) and junction potentials were kept constant. Figure 1 shows the electrode functions for the two electrodes. The responses are linear over the range 10-5-10-M with sub-Nernstian slopes (47 mV/ decade for electrode A and 48 mV/decade for electrode B). The slopes of the curved parts are only about 25 mV/decade for 10-5-10-6M solutions. Extrapolating to p[novocaine] = 0 gives 273 mV for E&&, and 292 mV for I$,,,, (both values LS. SCE).

Determination of novocaine


Table 1. Selectivity coefficients for the novocaine-selective membrane electrodes (CNorocrinc Cln,crler.nlL+= O.OlM; = pH = 5.0; I = 0.1 (NaNO,) Selectivity coefficient, log L.,j Electrode Electrode A B


Substances tested Leucine Ephedrine Benzoic acid Nicotinic acid Triethanolamine Piperazine Pyramidone Codeine Caffeine Atropine Urea Sodium nitrate (I = O.OlM) Disodium phosphate Potassium nitrate Potassium sulphate




1.70 4.53 4.32 3.15 4.06 1.17 1.25 2.91 0.51 4.51 5.72 5.55 3.89 3.68

4.54 2.04 4.27 3.75 2.79 3.56 1.00 1.13 2.70 0.78 4.15 4.48 3.70 3.92 3.56


data in Table 1 show that the K,,,,j for both electrodes.

Injhence of pH

values are similar

p CNovocainel
Fig. 1. The electrode functions of the novocaine-selective membrane electrodes: (a) electrode A; (b) electrode B (I = 0.1, NaN03 ; pH 5.0). Selectivity

Novocaine very often has to be determined in preparations in which it is mixed with organic and inorganic substances. These substances constitute a serious problem in the various existing methods of analysis and have to be adequately removed before the novocaine determination. The effect of some of these substances on the responses of our electrodes was estimated by measuring the selectivity coeffiby the method of Srinivasan and Rechcients, nitz17 and the equation:

The effect of pH on the potentials of the electrodes was examined by measuring the emf of the cell in novocaine hydrochloride solutions in which the pH was varied by adding appropriate amounts of hydrochloric acid and/or sodium hydroxide solution. At pH values between 2 and 6, no significant change in the membrane potential was observed (for the two electrodes and for different concentrations of novocaine hydrochloride). At pH values higher than 66.5, the novocaine base in the aqueous test solutions precipitates, so the emf values are shifted towards more negative values (Fig. 2).

log KnoV., j

-5, - 4,*

or 8)

+ log [novocaine+] (3)

- log [interferent+]l

where EItA,,rs) is the emf of cell (I) for c novocaine = O.OlM, E,t is the emf of an electrochemical cell of type (II) and S is the electrode slope. novocaineinterferent+ selective (C = O.OlM) membrane I = o,l (NaNo3) electrode (PI-I 5.0) (A or B) 1

satd. NaNO, , SCE solution I I



The results obtained with commonly met concomitants (Table 1) indicate that only atropine, codeine and pyramidone are likely to cause interference. The
TAL. 2816-c

Fig. 2. Influence of pH on the response of electrode A for different novocaine hydrochloride concentrations.



were about 3 min in the 10m6 and lo- M solutions, and about 30 set in 10m4M solutions. In more concentrated solutions the response order of a few seconds. times were of the


KIf2 M lPB Fig. 3. Potentiometric titration of 30 ml of lo-M novoCaine hydrochloride with O.OlM tetraphenylborate at pH 5.0: (a) electrode A; (h) electrode B. Response time

The electrode response times were measured for novocaine hydrochloride solutions of different concentrations. For both electrodes the response times

Both electrodes can be used in the potentiometric determination of novocaine both by direct potentiometry and by potentiometric titration. We found titration to be advantageous, especially in the case of determination of novocaine in complex phannaceutical preparations, because it is difficult to keep the constant ionic strength required for direct measurements. The most suitable titration reagent was found to be sodium tetraphenylborate (O.OlM aqueous solution or more dilute) which is stable for periods of up to a month. Figure 3 shows the potentiometric titration curves of 10e3M novocaine hydrochloride (pH 5.0, adjusted with hydrochloric acid and/or sodium hydroxide solution) obtained with the two electrodes. The obvious jumps of potential in both cases allow accurate evaluation of the equivalence volumes. In the determination of the active principle in the pharmaceutical preparations such as Gerovital and Aslavital (ampoules or tablets, composition given in Tables 2 and 3) both electrodes had almost similar behaviour. None of the additional ingredients affected the results of the determinations. In all cases, reproducibility was good. The mean relative error for 4-12 mg of novocaine hydrochloride does not exceed f 0.5%.

Table 2. Determination of novocaine hydrochloride in aqueous injectable solutions with a novocaine-selective membrane electrode* Pharmaceutical product Nominal amount,t W 4 8 12 12 8 12 12 12 12 12 12 i 12 12 12 F 12 12 12 12 Found. my 3.92 7.97 11.92 11.97 7.78 11.84 11.79 11.99 12.07 12.26 12.38 12.40 12.35 12.35 11.79 11.82 11.94 11.94 Found,

Composition 1 ampoule (5 ml) contains 100 mg of novocaine. HCI: 6 mg of benzoic acid; 5 mg of potassium metabisulphite; 1 mg of disodium hydrogen phosphate; distilled water to 5 ml 1 ampoule (5 ml) contains 100 mg of novocaine.HCI: 64 mg of glutamic acid: 6 mg of benzoic acid: 24 mg of potassium hydroxide: 5 mg of potassium metabisulphite; 0.5 mg of disodium hydrogen phosphate; distilled water to 5 ml


Gero HJ


98.0 99.7 99.4 99.5 97.3 98.7 98.3 99.9 100.6 102.8 103.3 103.4 103.1 103.1 98.3 98.5 99.5 99.5


II 111 IV

* Electrode A was used. t Assuming ampoule conforms exactly to specification,

Determination of novocaine Table 3. Determination of novocaine hydrochloride in tablets with a novocainet-selective membrane electrode* Nominal amount taken,t mg


Pharmaceutical product

Composition (per tablet) 100 mg of novocaine.HCI; 6 mg of benzoic acid; 5 mg of potassium metabisulphite; 0.5 mg disodium hydrogen phosphate; excipiens q.s. 100 mg of novocaine. HCI: 6 mg of benzoic acid; 5 mg of potassium metabisulphite: 0.5 mg of disodium hydrogen phosphate; 30 mg of vitamin B,,; 200 mg of meso-inositol ; excipiens q.s.

Found, mg 5.02 5.02 7.45 7.49 7.49 10.00 4.92 4.95 7.40 1.42 7.31 9.87

Found, mgl tablet 100.3 100.3 99.2 99.9 99.9 100.0 98.4 99.0 98.6 99.0 98.3 98.7

Gerovital Hs

5.0 1.5 7.5 7.5 10.0 5.0 5.0 7.5 7.5 7.5



* Electrode A was used. t Assuming tablets conform exactly to specification.

REFERENCES M. lonescu and V. V. Cosofret, Rec. Chim. (Bucharest). 1980, 31, 1005. Idem ibid., 1980, 31, 1088. G. E. Baiulescu and V. V. Cbsofret, Applications of
Ion-Selective Membrane Electrodes in Organic Analysis,

1975, XIX p. 409. 9. VS. Pharmacopeia. IO. Rom. Pharmacopeia, 1976, IX, p. 569. Il. E. Hopirtean and F. CSrmos, Stud. Univ. Babe?-Bolyai, Ser. Chem., 1977,22, 35. 12. C. Luca, C. Baloescu, G. Semenescu, T. Tolea and E.

Semenescu, Rev. Chim. (Bucharest),



Horwood, Chichester, 1977. _ 4. W. Selig, Ion-Selective Electrodes in Organic Elemental and Functional Group Analysis, Lawrence Livermore Laboratory, UCRL-52393, 1977 and 1978. 5. V. V. CosofreI, Ion Selective Electrode Rev., 1980, 2
(No. 4).


Cosofrel, P. G. Zug&escu and G. E. Baiulescu, Talanta, 1977, 24, 461. 14. V. V. Cogofrel, Reo. Roum. Chim., 1978, 23, 1489. 15. G. E. Baiulescu, V. V. Cogofrel and C. Christescu, Rev.
V. V. Chim., (Bucharest), 1975, 26, 429.

6. R. P. Buck, Anal. Chem., 1978, SO, l7R. 7. G. H. Fricke, ibid., 1980, 52, 259R. Vol. I, p. 367. Her Majestys 8. British Pharmacopoeia, Stationary Office London, 1980.

16. M. Ionescu, St. Cilianu, A. A. Bunaciu and V. V. Cogofret, Talanta, in the press. Il. K. Srinivasan and G. A. Rechnitz, Anal. Chem., 1969,
41, 1203.