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Surfactant Properties and Biodegradation of Polyethoxylates from Phenolic Lipids

John H.P. Tyman* and Ian E. Bruce

Department of Chemistry, Brunel University, Uxbridge, Middlesex, UB8 3PH United Kingdom

ABSTRACT: The surface tension of aqueous solutions of cardanol and cardol polyethoxylates derived from technical cashew nut-shell liquid were examined to determine their rel- ative surfactancy. The compositions with the greatest surface tension reduction, or optimal surfactancy, were selected for biodegradation testing, in terms of total organic carbon, using a soil inoculum. The experimental surfactants in this study were compared with commercial t-nonyl polyethoxylate and glucose as reference materials. Over a period of 28 d, car- danol polyethoxylate and its saturated analog were degraded to 17 and 25%, respectively, and cardol polyethoxylate and its saturated analog were degraded approximately half that ex- tent (37 and 46%). t-Nonylphenyl polyethoxylate remained substantially undegraded (77%), whereas glucose was almost completely degraded (5%). Paper no. S1373 in JSD 7, 169–173 (April 2004).

KEY WORDS:

of phenols, surface tension reduction, surfactancy.

Alkylphenol polyethoxylates, biodegradation

Technical cashew nut-shell liquid (CNSL), obtained by the industrial processing of phenolic lipids in the natural mate- rial Anacardium occidentale, which contains principally ana- cardic acid 1, cardol 2, and 2-methylcardol 3. Decarboxyla- tion gives cardanol 4, together with unchanged cardol, as shown in Scheme 1. The polyethoxylates 5 and 6, depicted in Scheme 2, were obtained by the reaction of cardanol and cardol with ethylene oxide (1). t-Nonylphenol polyethoxy- late 7 is shown for comparison. In each case, they contain a profile of oligomers in which m ranges from 1 to about 48. The ethoxylates were deemed ideal candidates for further surfactant and biodegradation studies, specifically, by first determining the optimal number of ethoxylate (EO) groups required for surfactancy, as judged by their relative reduction of the surface tension of water. With this infor- mation, biodegradation studies were carried out on the sample in each series exhibiting the highest surfactancy. Studies on the environmental occurrence of alkylphenols and alkylphenol ethoxylates (2) and their persistence in

*To whom correspondence should be addressed. E-mail: jhptyman@hotmail.com Abbreviations: CNSL, cashew nut-shell liquid; DOC, dissolved organic carbon; EO, ethoxylate; OECD, Organization for Economic Coopera- tion and Development; TOC, total organic carbon.

COPYRIGHT © 2004 BY AOCS PRESS

aquatic environments have both been reviewed extensively (3). Commercial alkylphenol polyethoxylates have received criticism on environmental and toxicological grounds (4), and although subsequent studies have suggested their accu- mulation is not significant (5), there also have been indica- tions that they could be gradually phased out (6). Recent work (7) has indicated the continued existence of marine pollution with alkylphenol polyethoxylates. Studies on the response of activated sludge to variations in the composition of feed effluent (8) have indicated that alkyl ethoxylate- adapted biomass was more capable of adaptation to alkylphenol ethoxylate than the reverse. Overall, there are conflicting views on the fate and effect of the present class of alkylphenol polyethoxylates in the environment. Since approximately 90% of the alkylphenol ethoxylates in the European Union are based on the petrochemical in- termediate t-nonylphenol, it is of interest to examine the use of natural alkylphenols, based on the supposition that a biosynthesized compound might prove to be more biodegradable and environmentally acceptable than t-nonylphenol and its polyethoxylates. Also, an ethoxylate from a natural alkylphenol would necessarily require useful surfactant properties, and in this regard, there has been lit- tle research on the surfactant activity of such products. As a result, both the surfactancy and biodegradability of the polyethoxylates from the phenolic lipids cardanol 4 and car- dol 2 were studied in the present work. The choice of biological medium—namely, whether to choose one based on activated sludge or one of soil origin— in the several test methods used for determination of the

in the several test methods used for determination of the SCHEME 1 JOURNAL OF SURFACTANTS AND

SCHEME 1

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170 J.H.P. TYMAN AND I.E. BRUCE SCHEME 2 biodegradability of polyethoxylates is important. In our work,

SCHEME 2

biodegradability of polyethoxylates is important. In our work, we elected to adopt a European Economic Commu- nity procedure based on a “shake flask” system rather than

to use activated sludge, as is generally applied in the United States. Some parameters of the Organization for Economic Cooperation and Development (OECD) screening test for the biodegradation of surfactants with regard to inoculum, aeration, feed, and media were examined; we concluded that no inoculum was needed to begin the biodegradation process and that it was possible to use standard mineral mi- croelements (9). Although the dilute bacterial media pres- ent in the shake flask systems do not simulate sewage treat- ment plant conditions, they are useful for the initial screening of a variety of substances and are adaptable to procedures involving radiolabeled materials.

A variety of chemical and spectroscopic techniques have

been used for the analytical determination of polyethoxy- lates. One of the many procedures in biodegradation stud- ies has been to determine the rate and extent of reduction of dissolved organic carbon (DOC), which involves a filtra-

tion step, on samples for analysis or those of total organic carbon (TOC), in which samples are not filtered. The most reliable and accurate method is probably the use of a car- bon analyzer such as an inductively coupled plasma spec- trometer. However, we found that, at the very low concen- trations used in our experiments, the errors encountered were too great. An alternative and simpler chemical method for determining TOC—one consisting of oxidation of the carbon present with acidic dichromate and absorption of the resulting carbon dioxide in barium hydroxide—was found to be satisfactory in this study. To determine biodegradability, a low concentration of microorganisms and a maximum duration of 28 d (de- signed to screen substances) was used. Next, TOC analyses were carried out on samples withdrawn periodically. Sub- stances passing this test with greater than 70% removal of DOC were assumed to be extensively degraded during sewage treatment in practice and were likely to be removed rapidly from bulk water.

It was of interest to compare the susceptibility of differ-

ent types of aromatic substitution in phenols described in re- cent biodegradation studies—often with a wide variety of specific organisms—with the phenolic structures of the poly- ethoxylates in the present work. Such work is of interest in charting the relationship between organic structure and the

biological activity of different classes of organisms. Thus, biodegradation studies with organic pollutants have been mainly concerned with bacteria, fungi, and yeasts. In an ex-

amination of other types of organic polluntants, namely, eu- karyotic algae such as Ochromonas danica (10), phenol was found to degrade more readily than p-cresol, although higher homologs were not examined. By contrast, with ther- mophilic microorganisms such as Bacillus thermoglucosidasius, nonoxidized aromatic compounds were not biodegraded, and phenol and m-cresol exhibited the highest degradation. This occurred to a much greater extent than the biodegra- dation of o- and p-cresol, (11). Even though the studies with this group of microorganisms have not included higher- chain-length compounds, the effect of m-substitution in the present phenolic polyethoxylates among other biological or- ganisms could nevertheless be of potential interest. In a study on the biodegradation of different phenols in river water (12) at mild temperatures, at neutral pH, and with excess oxygen, the relative order of biodegradation was found to be resorcinol > phenol > o-cresol > o-chlorophenol. The higher rate for 1,3-substitution may again have signifi- cance for longer-chain compounds. The 1,3,4, namely, partly m-substitution in lignans, was found to be susceptible (13) to a basidomycete, white rot fungus (Lentinula edodes). More generally, enzymatic systems have been investi- gated. Thus, the removal of phenolic pollutants in waste- waters by soybean peroxidase has been studied (14), and the advantages of enzyme-based, compared with physico- chemical, treatment have been indicated. The biodegrada- tion of phenol in coke wastewater has been examined with enzymes from Pseudomonas sp. immobilized on flat polyamide membranes (15). In studies reminiscent of the use of specific organisms to biotransform dedicated com- pounds to organic intermediates, the use of a recombinant strain of E. coli P260 has been examined for the controlled biodegradation of aromatic compounds and subsequent synthesis (16) of a variety of organic compounds. Some sources (17) have commented on the occurrence of phenolic and nonphenolic estrogenic mimics in the en- vironment. Indeed, the long-term effects upon the living world and the environment of the great range of present- day chemicals—very largely derived from petrochemical sources—remain relatively unknown. In this connection, the greater use of natural resources in chemical production (18) would appear to be timely.

EXPERIMENTAL PROCEDURES

Materials. Technical CNSL was obtained from 3M Research Ltd. (Harlow Essex, United Kingdom). The separation of car- danol and cardol from technical CNSL, their polyethoxylation

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with EO, and their characterization as surfactants have been described (1). t-Nonylphenol polyethoxylate, obtained from S.C. Johnson and Son, Inc. (Racine, WI) was of commercial origin. Reaction of the phenolic lipids, cardanol, and cardol (shown in Scheme 2) with EO was carried out in each case until an average of m = 48 EO groups/mol was added. In the characterization studies of these polyethoxylates, 16 samples were collected in the course of the polyethoxylation reactions and submitted to high-performance liquid chromatography and nuclear magnetic resonance spectroscopic examination. In each case, these same 16 samples were then also examined for their surfactant and biodegradation properties. The five polyethoxylates, namely, cardanol polyethoxylate 5 and its sat- urated analog 5 (n = 0), and cardol polyethoxylate 6 and its sat- urated analog 6 (n = 0), were first used to determine the species yielding the greatest reduction in the surface tension of water. Then, in each case, this optimal sample was submit- ted to biodegradation studies and compared with t-nonylphe- nol polyethoxylate 7. Surfactant property determinations. The reduction in the surface tension of distilled water (measured as Nm 1 ), by 1% aqueous solutions of cardanol and cardol polyethoxy- lates and by saturated versions of both polyethoxylates and of t-nonylphenol polyethoxylate, was determined at 24°C by the use of a du Noüy tensiometer (19). In this method a cir- cular ring of radius r attached by a single wire at diametri- cally opposite points rests horizontally and is completely wetted on the surface of the solutes. The force, f, required to detach it from the opposing vertical action of the surface tension, γ (at zero contact angle), is given by f = 4πr γ. The equipment is operated manually. Biodegradation experiments. The method used was based on a modified OECD screening test for biodegradability (20), an alternative approach to using activated sludge. The test compound, in a standard nutrient medium at a concentra- tion of 100 mg/L, was inoculated with a low concentration of microorganisms from a mixed population and aerated at constant temperature for no more than 28 d. Biodegradation was measured by TOC analysis. Control flasks were run in parallel to determine TOC blanks, and the procedure was checked in duplicate with a reference compound of known biodegradability, namely, glucose at 100 mg/L. (i) Inoculation. Soil (100 g) (fertile, not sterilized, and free of herbicides and fertilizers) was suspended in water (1000 mL) and stirred, and the suspension was allowed to settle for 30 min. The supernatant liquid was filtered through coarse filter paper, the first 200 mL was discarded, and the remainder was aerated immediately until used. The inoculum was used on the day of collection. Materials and reference substances were evaluated in duplicate. The nutrient solution (200 cm 3 ) containing the required TOC concentration of the test compound (100 mg/L) was treated with the inoculum (0.1 cm 3 ). Controls with no test substance but containing the inoculum were also run. During the experiment, a suitable volume (10 cm 3 ) of

PROPERTIES AND BIODEGRADATION OF POLYETHOXYLATES

the test material, the reference, and the control were with- drawn in each case, but not filtered, and the TOC was de- termined in duplicate from samples in two reaction flasks fitted with loose metal caps that did not impede airflow. The flasks were gently agitated in a shaker shielded from light for 28 d at 25°C, and samples were withdrawn for TOC determination every 4 d. (ii) TOC analysis. The acidic dichromate method was used (21). The carbon dioxide that evolved was trapped by standard aqueous barium hydroxide, which was then titrated with standard acid.

RESULTS AND DISCUSSION

Surfactancy. Although surfactancy may be assessed in a vari- ety of ways, such as by measuring the foaming ability of aqueous solutions, or their oil-solubilization characteristics, the reduction in surface tension (Nm 1 ) of water by dilute solutions of polyethoxylates was used in this study to deter- mine the optimal average ethoxylate chain length of the var- ious samples. The particular ethoxylate sample in each of the series yielding the greatest reduction of surface tension was then considered the best candidate for a biodegrada- tion study. The reductions in surface tension of 1% solutions of car- danol, cardol, and 3-pentadecylphenol polyethoxylates, re- spectively, are shown in Figure 1. For purposes of compari- son, a 1% solution of common commercial t-nonylphenol polyethoxylate 7, with an average of 8 EO groups, gave a reduction in surface tension of 33 Nm 1 . From these results, the samples of cardanol polyethoxy- late 5, cardol polyethoxylate 6, and 3-pentadecyl polyethoxy- late 5 (n = 0), with an average of 13.4 EO, 10.3 EO, and 14.8 EO groups, respectively, exhibited the greatest reductions in surface tension among the 16 samples examined in each class. Cardanol polyethoxylate and its saturated analog ap- peared to be similar in surfactancy, as expected, with side- chain unsaturation conferring slightly improved properties.

chain unsaturation conferring slightly improved properties. FIG. 1. Variation in the reduction in surface tension of

FIG. 1. Variation in the reduction in surface tension of water by car- danol, 3-pentadecylphenol, and cardol polyethoxylates with the aver- age number of ethylene oxide (EO) groups/mole.

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By contrast, cardol polyethoxylate 5 exhibited a relatively smaller reduction in surface tension and, consequently, would be expected to be a less effective surfactant. Although technical CNSL itself might prove a useful surfactant upon polyethoxylation, the removal of cardol appears to be bene- ficial for achieving the highest potential surfactancy. A comparison of the three compounds depicted in Fig- ure 1 revealed that t-nonylphenol polyethoxylate 7 was com- parable in surfactancy with cardanol polyethoxylate and that the sample with an average of 8 EO groups was consid- ered to be an appropriate reference material in biodegra- dation work. Biodegradation studies. As shown in Figures 2 and 3, the per- centages of initial TOC of cardanol polyethoxylate and 3-pen- tadecylphenol polyethoxylate remaining after 28 d were 17 and 25%, respectively, compared with 77% for t-nonyl poly- ethoxylate. The biodegradation of cardanol polyethoxylate was comparable to that of the reference material glucose. In contrast, the corresponding results for cardol poly- ethoxylate and saturated cardol polyethoxylate were 37 and

ethoxylate and saturated cardol polyethoxylate were 37 and FIG. 2. Biodegradation of cardanol, 3-pentadecyl phenol,

FIG. 2. Biodegradation of cardanol, 3-pentadecyl phenol, and t- nonylphenol polyethoxylates. TOC, total organic carbon; for other ab- breviation see Figure 1.

organic carbon; for other ab- breviation see Figure 1. FIG. 3. Biodegradation of cardol and saturated

FIG. 3. Biodegradation of cardol and saturated cardol polyethoxy- lates and glucose. For abbreviations see Figures 1 and 2.

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46%, respectively, confirming that linear hydrophobic chains are highly beneficial to biodegradation and that cer- tain types of unsaturation appear advantageous. Our results are in agreement with earlier studies showing that certain types of branching in the chain of alkyl polyethoxylates have a negative effect on their biodegradability. Also, the pres- ence of two ethoxylate side chains in cardol polyethoxylate might partly explain its slower biodegradation compared with that of cardanol polyethoxylate. The inferior surfac- tancy and biodegradability of cardol polyethoxylate, as com- pared with cardanol polyethoxylate, would seem to justify separation of the component phenols in technical CNSL prior to ethoxylation. A cost/benefit study covering the full range of uses of these polyethoxylates could prove useful in this regard, bearing in mind that technical CNSL from dif- ferent regional sources contains approximately 20% cardol. The nature of the organic material constituting the TOC after 28 d has not been identified. By analogy with other work (22), it seems probable that it constitutes the car- boxymethyl derivative of cardanol, cardanol itself, or fur- ther metabolic degradation products. For t-nonylphenol polyethoxylate, biological degradation of the side chain by both aerobic and anaerobic pathways has been described (3, p. 41), leading to t-nonylphenoxyacetic acid from 2-t- nonylphenoxy ethanol. One could reasonably conclude that natural phenolic lipids behave similarly. Although it would indeed be of interest to identify the metabolites by high-performance liquid chromatography–mass spectrome- try or other techniques, this analysis was not performed in the present work. Concern has been expressed regarding the use of t-nonyl polyethoxylates because of their slow and incomplete biodegradation. The surfactants derived from natural cashew phenols, having improved biological compatibility as described in the present work, would appear to be an at- tractive alternative class of surfactants.

ACKNOWLEDGMENTS

We thank S.C. Johnson and Son Inc. for financial support. Dr. S. Smith, Department of Biology, is thanked for help with the biodegra- dation experiments. This paper is part 37b of the Long-Chain Phe- nols series. Part 37a appeared as “Synthesis and Characterization of Polyethoxylate Surfactants Derived from Phenolic Lipids” (JSD 6, 291–297, 2003).

REFERENCES

1. Tyman, J.H.P., and I.E. Bruce, Synthesis and Characterization of Polyethoxylate Surfactants Derived from Phenolic Lipids, J. Surfact. Deterg. 6:291 (2003).

2. Bennie, D.T., Review of the Environmental Occurrence of Alkylphenols and Alkylphenol Ethoxylates, Water Qual. Res. J. Canada 34:79 (1999).

3. Maguire, R.J., Review of the Persistence of Nonylphenol and Nonylphenol Ethoxylates in Aquatic Environments, Water Qual. Res. J. Canada 34:37 (1999).

4. Marcomini, A., B. Pavoni, A. Sfriso, and A.A. Orio, Persistent Metabolites of Alkylphenol Polyethoxylates in the Marine En- vironment, Chemosphere 17:853 (1988).

173

PROPERTIES AND BIODEGRADATION OF POLYETHOXYLATES

5.

Naylor, C.G., Environmental Fate of Alkylphenol Ethoxylates, Soap Cosmet. Chem. Spec. (August 27, 1992).

15. Bohdziewicz, J., Biodegradation of Phenols by Enzymes from Pseudomonas sp. Immobilised onto Ultracentrifuge Mem-

16. Ferror Munoz, E., and A. Gibello Prieto, Process for the

6.

Birtwistle, J.S., The Global Outlook for Surfactants from a U.S. Perspective, paper presented at the Fourth World Surfactants Congress (CESIO), Barcelona, Spain (June 1996).

branes, Process Biochem. (Oxford) 33:811 (1998).

Biodegradation of Aromatic Compounds and Synthesis of Pig-

7.

Zoller, U., and M. Hushan, The Non-ionic Surfactant Pollution Profile of Israel Mediterranean Sea Coastal Water, Water Sci. Technol. 43:245 (2001).

ments, Colorants, Alcaloids, and Polymers with the Use of the Recombinant Strain Escherichia coli P260, European Patent 0857779 (1998).

8.

Carvalho, G., E. Paul, J.M. Novais, and H.M. Pinheiro, Studies on Activated Sludge Response to Variations in the Composi- tion of a Synthetic Surfactant-Containing Feed Effluent, Water

Bryja, M., N. Howaniec, S. Nocon, I. Szwach, and M. Janosz-Ra-

Semple, R.T., Biodegradation of Phenols by a Eukaryotic Alga,

17. Sumpter, J.P., S. Jobling, and C.R. Tyler, Estrogenic Substances in the Aquatic Environment and Their Potential Impact on An- imals, Particularly Fish, Soc. Exp. Biol. Semin. Ser. 57:205 (1996).

9.

Sci. Technol. 42:135 (2000).

18. Tyman, J.H.P., The Role of Biological Raw Materials in Synthe- sis, in Studies in Natural Products Chemistry, edited by Atta-ur-

19. Taylor, H.S., and A.T. Taylor, Elementary Physical Chemistry, 3rd

20. Council Directive of March 31st, 1982, on the Approximation

10.

jczyk, Biodegradation of Surfactants According to the OECD Screening Test, Inz. Ochr. Srodowiska 1:215 (1998).

Res. Microbiol. 148:365 (1997).

Rahman, Elsevier, New York, 1995, Vol. 17, p. 601.

edn., Van Nostrand, New York, 1944, p. 140.

11.

Bauer, M.P., F.M. Duffrer, and R. Mueller, Biodegradation and Biotransformations of Cresols and Phenols by Thermophilic Microorganisms, DECHEMA–Monogr. Biodeterior. Biodegrad. 133 :575 (1996).

of the Laws of the Member States Relating to Methods of Test- ing the Biodegradability of Nonionic Surfactants and Amend- ing Directive 73/404/EEC/82/242.EEC, Official J. Eur. Com- mun. 109 (1982).

12.

Zhao, J., K. Zhang, C. Li, and P. Luo, Biodegradation of Phe- nols in River Water, Huaxi Yike Daxue Xuebao 31:367 (2000).

21. Huddleston, R.L., and R.C. Alldred, Evaluation of Detergent by Using Activated Sludge, J. Am. Oil Chem. Soc. 41:732 (1964).

13.

Giovannozzi-Sermani, G., A. D’Annibale, C. Crestini, and R.S. Stazi, Biodegradation of Aromatic Compounds by a White Rot Fungus, Recent Res. Dev. Biotechnol. Bioeng. 1:191 (1998).

22. Watson, J.S., D.M. Jones, and R.P.J. Swannell, Formation of Carboxylic Acids During Biodegradation of Crude Oil, In situ Bioremediation of Petroleum Hydrocarbons and Other Organic Com-

[Received May 21, 2003; accepted November 12, 2003]

14.

Wright, H., and J.A. Nicell, Removal of Phenolic Compounds

pounds, edited by B.C. Allerman and A. Leeson, Batelle Press,

from Waste Waters Using Soyabean Peroxidase, in Proceedings of the 50th Industrial Waste Conference, May 8–10, 1995, edited by R.F. Wukasch and C.S. Dalton, Lewis Publishers, Chelsea, MI, 1997, p. 495.

Columbus, OH, 1999, p. 239.

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