COLLAGEN EVALUATION 341 (9) Standard Methods for the Examination of Water and Wastewater (American Public Health Association, American Water Works Association and Water Environment Federation, Washington, D.C., 1995). (10) E. J. Miller and R. K. Rhodes, Preparation and characterization of the different types of collagen, Meth. Enzymol., 82, 33-64 (1982). (11) R. L. Trelstad, V. M. Catanese, and D. F. Rubin, Collagen fractionation: Separation of native types I, II and III by differential precipitation, Anal. Biochem., 71, 114-118 (1976). (12) U. K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227, 680-685 (1970). (13) B. Sykes, B. Puddle, M. Francis, and R. Smith, The estimation of two collagens from human dermis by interrupted gel electrophoresis, Biochem. Biophys. Res. Commun., 72, 1472-1480 (1976). (14) J. A. M. Ramshaw and J. A. Werkmeister, Electrophoresis and electroblotting of native collagens, Anal. Biochem., 168, 82-87 (1988). (15) T. Mclellan, Electrophoresis buffers for polyacrylamide gels at various pH, Anal. Biochem, 126, 94-99 (1982). (16) A.G. Gornall, C. J. Bordawill, and M. M. David, Determination of serum proteins by means of the biuret re�ction,J. Biol. Chem., 177, 751-766 (1949). (1 7) K. Kadlec, Extracellular matrix. 1. Fibril-forming collagens, Protein Profile, 5, 519-638 (1994). (18) G. Huszar, J. Maiocco, and F. Naftolin, Monitoring of collagen and collagen fragments in chroma­ tography of protein mixtures, Anal. Biochem., 105, 424-429 (1980). (19) K. J. Payne and A. Veis, Fourier transform IR spectroscopy of collagen and gelatin solutions: Decon­ volution of the amide I band for conformational studies, Biopolymers, 27, 1749-1760 (1988). (20) M. Nagelschmidt and B. Viell, Polarimetric assay for the determination of the native collagen content of soluble collagen,]. Biomed. Mater. Res., 21, 201-209 (1987). (21) T. Hyashi, S. Curran-Patel, and D. J. Prockop, Thermal stability of the triple helix of type I procol­ lagen and collagen: Precautions for minimizing ultraviolet damage to proteins during circular dichro­ ism studies, Biochemistry, 18, 4182-4187 (1979). (22) J. A. M. Ramshaw, Distribution of type III collagen in bovine skin of various ages, Connective Tissue Res., 14, 307-314 (1986). (23) R. A. Condell, V. P. Hanko, E. A. Larenas, G. Wallace, and K. A. McCullough, Analysis of native collagen monomers and oligomers by size-exclusion high-performance liquid chromatography and its application, Anal. Biochem., 212, 436-445 (1993). (24) J. E. Eastoe and A. A. Leach, "Chemical Composition of Gelatin," in The Science and Technology of Gelatin, A.G. Ward and A. Courts, Eds. (Academic Press, London, 1977), pp. 73-107. (25) C. C. Danielsen, Precision method to determine denaturation temperature of collagen using ultraviolet difference spectroscopy, Coll. Relat. Res., 2, 143-150 (1982). (26) P. Bruckner, and D. J. Prockop, Proteolytic enzymes as probes for the triple-helical conformation of procollagen, Anal. Biochem., 110, 360-368 (1981). (27) W. Friess and G. Lee, Basic thermoanalytical studies of insoluble collagen matrices, Biomaterials, 17, 2289-2294 (1996). (28) D. G. Wallace, R. A. Condell, J. W. Donovan, A. Paivinen, W. M. Rhee, and S. B. Wade, Multiple denaturational transitions in fibrillar collagen, Biopolymers, 25, 1875-1895 (1986). (29) J. A. M. Ramshaw, J. A. Werkmeister, and H. A. Bremner, Characterization of type I collagen from the skin of blue grenadier (Macruronus novaezelandiae), Arch. Biochem. Biophys., 267, 497-202 ( 1988). (30) H. C. Wang, J. D. Chen, G. C. Li, and F. H. Yew, Characterization of a heat-resistant strain of Tilapia ovary cells,]. Cell Sci., 92, 353-359 (1989). (31) N. K. Shah, M. Sharma, A. Kirkpatrick, J. A. M. Ramshaw, and B. Brodsky, Gly-Gly-containing triplets of low stability adjacent to a type III collagen epitope, Biochemistry, 36, 5878-5883 (1997). (32) M. Chvapil, Collagen in cosmetics: Misconception of its mode of action, Cosmet. Toiletr., 97, 35-36 (1982). (33) S. D. Coapman, J. L. Lichtin, A. Sakr, and J. R. Schiltz, Studies of the penetration of native collagen, collagen alpha chains, and collagen cyanogen bromide peptides through hairless mouse skin in vitro, J. Soc. Cosmet. Chem., 39, 275-281 (1988).

J. Cosmet. Sci., 55, 343-350 CTuly/August 2004) Quantitative determination of formaldehyde in cosmetics using combined headspace-solid-phase microextraction­ gas chromatography RENE THOMAS RIVERO and VINOD TOPIW ALA, Coty Research and Development Center, 410 American Road, Morris Plains, NJ 07950-2451. Accepted for publication May 21, 2004. Synopsis The objective of this research was the application of headspace (HS)-solid-phase microextraction (SPME) for the quantitation of formaldehyde present in raw materials and cosmetic formulations. The formaldehyde was derivatized in situ first with pentafluorophenylhydrazine (PFPH), to form a derivative hydrazone. The formed hydrozone was adsorbed on a SPME fiber during headspace extraction under controlled conditions (time, temperature, volume, etc.). After the adsorption step, the SPME fiber was directly transferred into the gas chromatography (GC) injection port in which the analytes were thermally desorbed. Deuterated acetone was used as an internal standard (IS) in order to quantitate the formaldehyde content. For the experiment, a gas chromatograph equipped with a flame ionization detector (GC/FID) was employed. A gas chromato­ graph/mass spectrometer (GC/MS) was used for the qualitative confirmation of results in this work. INTRODUCTION For decades, formaldehyde was one of the most widely used preservatives in personal care products due to its versatility, cost, and efficiency. In recent years, the potential for carcinogenic and respiratory sensitization from formaldehyde has become widely under­ stood. This has led to a movement in the industry that has imposed regulations, restrictions, and formaldehyde's usage being banned. In surfactant and cosmetic industries several analytical procedures have been developed for qualitative as well as quantitative analysis. One of the most well known procedures for qualitative analysis is the phloroglucinol test, in which formaldehyde reacts in an alkaline medium with phloroglucinol to produce a reddish-brown color complex. For the quantitation of formaldehyde, several methods have been employed, one of which is based on Nash reagent. In this determination, formaldehyde is condensed with ammonia and acetylacetone to form the lutidine derivative 3,5-diacetyl-1,4-dihydrolutidine (1,2). Nash reagent is sensitive not only to formaldehyde, but also to other aldehydes. If chromophore compounds are present in the product, they could interfere with the spectrophotometric determination. Formaldehyde can also be determined colorimetri- 343