sodium monofluorophosphate in oral hygiene products
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1 j. Cosmet. Sci., 52, (November/December 2001) Simultaneous quantitative determination of fluorine and sodium monofluorophosphate in oral hygiene products LAI-HAO WANG, Department of Applied Chemistry, Chia Nan University of Pharmacy and Science, Tainan, Taiwan 7171 O, R.O.C. Accepted for publication August 15, Synopsis An ion chromatographic method for simultaneous quantitative determination of fluorine and sodium monofluorophosphate in oral hygiene products is described. The liquid chromatographic system consisted of an IC A1 polymethacrylate-based anion exchanger and carbonate buffer (ph 9.85) as the mobile phase with a conductive detector. Various excipient ions were investigated with respecto their interference with the determination of fluoride. Comparison with results obtained from a fluoride-ion electrode technique show good agreement. INTRODUCTION Epidemiologic and experimental evidence concerned with the relationship of fluoride to dental caries suggests that fluoride solutions applied to the external surfaces of teeth may decrease their susceptibility to caries. The first two papers on the effect of topical application of fluorides were made by Bibby in 1942 (1) and Chenye in 1946 (2). Fluorine derivatives including sodium fluoride, stannous fluoride, and sodium monofluorophosphate (MFP, Na2PO3F) are incorporated into dentifrices or mouthwashes chemotherapeutic agents (3). However, fluoride is absorbed into the blood from the gastrointestinal tract. It is then mostly deposited in bone or excreted in urine (4). The acute and chronic toxicity values (LD5o values in the rat and mouse) of sodium fluoride, stannous fluoride, and MFP have been investigated. The toxicity of stannous fluoride is similar to that of sodium fluoride. Stannous fluoride and sodium fluoride are more toxic than MFP by factors of (4). The Food and Drug Administration (FDA) has promulgated regulations for safe and effective oral hygiene products. Active anticaries agents in dentifrices were 0.22%, 0.40% and 0.76% for sodium fluoride, stannous fluoride, and MFP, respectively. The final product, containing 0.02% fluoride ion in oral rinses, was based on in vitro data as well as on clinical trials (4). A mutagenicity study on three fluorine derivative selected from the cosmetic guidelines 399
2 400 JOURNAL OF COSMETIC SCIENCE of the Council of the European Communities (27 July 1976) was published (5). Stannous fluoride was slightly mutagenic in the Ames test. Today, over 90% of the dentifrices sold in Taiwan contain fluoride from one of two major sources (sodium fluoride and MFP). There are reports that a mixture of sodium fluoride and MFP is superior in efficacy to MFP alone in a dentifrice base (4). Dentifrices with mixed fluoride systems been marked. Quantitative determination of fluoride and MFP are important for quality control and stability evaluation of these products. Gas chromatographic (6-8), fluoride-ion electrode (6,8,9) high-performance liquid chromatographic (10), and ion chromatographic (11,12) techniques have been used to determine fluoride derivatives in toothpastes. Gas chromatographic methods involve the chromatographic analysis of trimethyl fluorosilane resulting from the reaction of trimethyl chlorosilane with fluoride ions in the toothpaste. A fluoride-ion electrode technique was used only for the determination of soluble fluoride derivatives. However, MFP is soluble in water to the extent of 42% saturation and is slowly hydrolyzed in the presence of hydrochloric acid. These methods are laborious and time-consuming. The use of high-performance liquid chromatography required a postcolumn detection system in order to analyze orthophosphate, diphosphate, triphosphate, and cyclotriphosphate in MFP samples (10). Ion chromatographic methods for direct determination of MFP in toothpaste have been reported (11,12). However, oral hygiene formulations commonly contain abrasives (phosphate), an antibacterial agent (cetylpyridium chloride), astringent salts (zinc chloride), and a surfactant (sodium lauryl sulfate). These excipient anion ions were found not to be ideal for resolving fluoride and phosphate at ph This present paper describes the application of various ph to separate rapidly and efficiently the peaks of interest. The results were compared with those obtained with the fluoride- ion electrode method. EXPERIMENTAL ION CHROMATOGRAPHY The liquid chromatograph was the Shimadzu LC-10 AD chromatography module containing the pump and conductivity detector (Shimadzu CDD-6A). The analytical column used a Shim-Pack IC-A1 (4.6 mm ID x 10 cm) polymethacrylate-based anion exchanger (10 l m). Chromatographic data were collected and analyzed with a Chromatopac C-R6A. The following chromatographic conditions were used: eluent flow rate, 1.5 ml min - ' conductivity temperature, 40øC; sensitivity, 1.0 l s cm - ' injection volume, 50 lal; recorder chart speed, 5 mm/min - All reagents used were analytical grade. The MFP was obtained from Aldich Chemical Corporation and had an assay value of 95%. The ph of the eluent, containing 0.94 m mol 1 - sodium carbonate, was adjusted to 9.85 and with 0.31 m mol 1 - sodium hydrogen carbonate and 1.0 mol 1-1 sodium hydroxide, respectively. The standard fluoride (1000 mg 1- ), chloride (1000 mg l-i), sulfate (1000 mg 1- ), and MFP (0.1%) solutions were prepared by dissolving the appropriate amounts of sodium salt in double deionized water. A 500-mg amount of toothpastes or mouthwashes was accurately weighed into a beaker, and 20 ml of deionized water was added and stirred until the sample was fully dispersed.
3 FLUORIDES IN ORAL HYGIENE PRODUCTS 401 After centrifugation, the supernatant liquid was transferred into a 50-ml volumetric flask and made up to volume with double aleionized water. An aliquot of the dispersion was filtered through a 0.2-pro (Gelman Sciences, Super Acrodisc) filter for analysis. FLUORIDE-ION ELECTRODE TECHNIQUE An amount of 2.0 g of the sample toothpastes or mouthwashes was weighed into a 50-ml beaker, and 6 ml of deionzied water was added and stirred until the sample was fully dispersed. After centrifugation, the supernatant liquid, after the treatment with hydro- chloric acid to hydrolyze the FPO3 2- ions, was made up to volume with deionized water. A 1.0-ml aliquot of the acid liquid (soluble fluoride) was neutralized with 4 tool 1-1 sodium hydroxide. The neutralized solution of sample was mixed with sodium acetate buffer (ph 4.82) containing 0.1% CDTA [(+)trans-l,2-diaminocyclohexane- N,N,N',N'-tetraacetic acid monohydrate] or TISAB I (0.50 tool 1 - sodium chloride, 0.25 mol 1-1 trisodium citrate, and 0.50 tool 1-1 acetic acid) or TISAB II (1.83 mol 1-1 sodium acetate and tool 1 - citric acid). The soluble fluoride concentration was determined with an Orion Research model fluoride-ion electrode and an Orion model 420 mv meter. Calibration graphs were constructed using a standard solution of sodium fluoride. RESULTS AND DISCUSSION OPTIMIZATION OF THE MOBILE PHASE Dentifrices or mouthwashes containing anions include the fluoride, MFP, phosphate, and chloride in many commercial ingredients, and sulfate as an impurity in some anionic surfactants. The separation mixture of the anions of the short run times is difficult. Separation and quantification of fluoride and monofluorophosphate (FPO3 2-) was interference-free in the wide range of separation conditions selected. At the elevated mobile phase ph value, an HPO42- peak was shifted away from the FPO32- peak. Carbonate buffer with ph gave good separations of F- and FPO32-, but the overall analysis time was longer than with ph The carbonate buffer (ph 9.85) contained a mixture of 0.94 m mole Na2CO 3 and 0.31 m mole NaHCO 3. The CO32- concentration helped achieve a largeresolution between FPO32- and SO42-. Chloride was also eluted in a chromatographic region free of interference from other ion peaks. The retention times were 2.683, 5.448, 8.473, and rain for F-, CI-, FPO3 2-, and SO42-, respectively. Optimization of CO32- concentration and the eluent ph for maximum resolution at minimum total run time resulted in the eluent described in the Experimental section. A chromatogram of a standard solution mixture produced with this eluent is shown in Figure 1. Under these condition separation was fast, reproducible, and free from interference from other components of the sample. REPRODUCIBILITY AND ACCURACY Determination of the concentratin of the various chemotherapeutic agents was accomplished by means of a calibration graph. The calibration graphs were linear for two chemotherapeutic agents over the range of concentration used ( mg l-p). The correlation coefficients were within the range of Recovery tests were
4 402 JOURNAL OF COSMETIC SCIENCE I ) i..j Time( min ) Figure 1. Chromatogram of standard solution using sodium carbonate buffer (ph 9.85). Mobile phase. Peaks: a = fluoride (25 mg/1); b = monofluorophosphate (30 mg/1). carried out on oral hygiene products to evaluate the reproducibility and accuracy of the proposed ion chromatographic (IC) method. Four toothpastes and mouthwashes were spiked with the amounts reported in Table I and subjected to the whole procedure. As
5 FLUORIDES IN ORAL HYGIENE PRODUCTS 403 Table 1. Recovery and Reproducibility in the Analysis of Toothpastes and Mouthwashes by Ion Chromatography (5 determinations for each aspect of procedure) Sodium fluoride Sodium monofluorophosphate Added Found Recovery Added Found Recovery (mg 1 - ) (mg 1 ) (%) (mg 1 t) (mg 1 - ) (%) Toothpaste A (1.4) a (2.3) Toothpaste B (5.0) Mouthwashes A (3.3) Mouthwashes B (1.3) Standard deviation. shown in Table I, excellent recoveries and precision were observed (recoveries ranging from 97.3% to 103.5%). APPLICATION TO ORAL HYGIENE PRODUCTS The proposed IC method was applied to the determination of chemotherapeutic agents in oral hygiene products (toothpastes and mouthwashes). A representative chromatogram of a commercial toothpaste is shown in Figure 2. Analytical results are given in Time( min ) Figure 2. Chromatogram of typical toothpaste using sodium carbonate buffer (ph 9.85). Mobile phase. Peaks: a = fluoride; b = rnonofluorophosphate.
6 404 JOURNAL OF COSMETIC SCIENCE Table Analytical Results for the Determination of Fluoride and Monofiuorophosphate in Commercial Toothpastes and Mouthwashes by Ion Chromatography (IC) and Fluoride-Ion Selective Electrode Concentration (w/w, %) II IC (n = 5 ) F- electrode (n = 6 ) F- FPO32 Total F- Toothpaste (4.3%) b (2.5%) Toothpaste (1.1%) (1.4%) (5.0%) Toothpaste (0.7%) (2.4%) (2.2%) Toothpaste (3.3%) (4.1%) (2.3%) Toothpaste (5.0%) (4.0%) (0.5%) Toothpaste (3.7%) (4.4%) (4.4%) Toothpaste (2.0%) (4.4%) (0.76%) Toothpaste 8 0, (0.9%) (5.1%) (0.7%) Toothpaste (2.4%) (3.7%) (1.8%) Toothpaste (2.5%) (3.9%) Toothpaste (3.6%) (5.6%) Toothpaste (5.3%) (5.5%) (1.6%) Mouthwash (1.2%) (1.5%) Mouthwash (2.8%) (2.6%) Number of determinations. Relative standard deviation. Table II. These results agree with those obtained by a fluoride-ion electrode method, performed with a TISAB II solution, after treatment with hydrochloride acid hydrolysis. CONCLUSIONS The IC procedures described here are applied directly to the analysis of cosmetic samples without the need for MFP acid hydrolysis. The direct determination of fluoride not only offers more precision than indirect determination but also saves more time and is applicable to actual samples. ACKNOWLEDGMENT Financial support for this work by the National Science Council of the Republic of China is gratefully acknowledged (no. NSC M ).
7 FLUORIDES IN ORAL HYGIENE PRODUCTS 405 REFERENCES (1) B.G. Bibby, A new approach to caries prophylaxis: A preliminary report on the use of fluoride application, Tufts Dental Outlook, 6, 4 (May 1942). (2) V. D. Cheyne, Human dental caries and topically applied fluorine: A preliminary report,j. Am. Dent. Assoc., 29, 1683 (1946). (3) M. V. Verling, "Dentifrice," in The Chemistry and Manufacture of Cosmetics, 2nd ed., M. G. alenavarre, Ed. (Continental Press, Orlando, FL, 1975), Vol. 2, pp (4) M. Pader, Oral Hygiene Prod/acts and Practice (Marcel Dekker, New York, 1988), pp (5) E. Gocke, M. T. King, K. Eckhardt, and D. Wild, Mutagenicity of cosmetics ingredients licensed by the European Communities, Mutat. Res., 90, (1981). (6) D.C. Wayne, An investigation of methods proposed for the analysis of the various forms of fluoride in toothpaste, Aust. Dent. J, 26, (1981). (7) M. S. Ma and W.J. Chen, Determiantion of active fluoride content in toothpaste by gas chromatography, Riyong Huaxue Gongh. 2, (1988). (8) S. Emil, P. Anica, K. Adolf, and P. Mirko, Quantitative determination of fluoride in toothpastes, J. High. Resoh#. Chromatogr., 16, (1993). (9) D. Y. Liu, Determination of fluoride content in toothpaste by ion-selectivelectrode, Riyong Huaxue Gongh, 3, (1992). (10) Y. Norimass, N. Sachiko, N. Tetsuya, U. Nobuyuki, K. Hiroki, and T. Akira, Determination of monofluorophosphate, orthophosphate, and polyphosphates by high-performance liquid chromatography with a photodiode array detector, Anal. Chem., 64, (1992). (11) J.P. Jeffrey, E. H Audrey, and J.B. Geoffrey, Determination of fluoride and monofluorophosphate in toothpaste by ion chromatography, J. Chromatogr., 367, (1986). (12) H. T. Jeseph and A. B. Thomas, Determination of potassium nitrate and sodium monofluorophosphate in the presence of phosphate and sulfate by high-resolution ion chromatography. J. Chromatogr., 410, (1987).
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