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Talanta 74 (2008) 13711377

On-line pretreatment and determination of parabens in cosmetic products by combination of ow injection analysis, solid-phase extraction and micellar electrokinetic chromatography
Fang Han, You-Zhao He , Chang-Zhu Yu
Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, China Received 2 July 2007; received in revised form 12 September 2007; accepted 13 September 2007 Available online 19 September 2007

Abstract A convenient and automated method for on-line pretreatment and determination of three parabens (i.e. methyl, ethyl and propyl phydroxybenzoate) in cosmetic products is proposed by using ow injection analysis (FIA), solid-phase extraction (SPE) and micellar electrokinetic chromatography (MEKC). An improved splitow interface is used to couple SPE on C8 -bonded silica with MEKC separation, which can avoid running buffer contamination and reduce buffer consumption, especially, containing expensive reagents. The analytes are loaded onto a C8 column at 0.6 mL/min for 60 s and eluted with a mixed eluent of 40% (v/v) 10 mmol/L sodium tetraborate buffer (pH 9.3) and 60% (v/v) ethanol at 0.75 mL/min. The MEKC separation is accomplished with a running buffer of 20 mmol/L sodium tetraborate (pH 9.3) containing 100 mmol/L sodium dodecyl sulfate (SDS) at 15 kV. For butyl p-hydroxybenzoate did not be detected in the cosmetic products, it was used as an internal standard (IS) added into the real samples. This FIASPEMEKC method using IS allows the sample separation within 12 min and the sample throughput of ve samples per hour with the relative standard deviation (R.S.D.) less than 2.3% (n = 5). The limits of detection (LOD) are in the range from 0.07 to 0.1 g/mL (S/N = 3 and n = 11). The proposed method has been used to determine three parabens in real cosmetic products satisfactorily. 2007 Elsevier B.V. All rights reserved.
Keywords: Flow injection analysis; Solid-phase extraction; Micellar electrokinetic chromatography; Splitow interface; Parabens

1. Introduction Parabens, viz., alkyl p-hydroxybenzoates, are widely used as preservatives in cosmetic products due to their broad spectrum of antimicrobial activity and ideal physicochemical properties [13]. Several animal and human studies on the safety of parabens appeared in recently published papers indicate that the exposure to parabens may modulate and disrupt the endocrine system, and may have harmful effects on human health [4,5]. Therefore, the highest concentration of single paraben allowed in a cosmetic product is 0.4% (w/w), and the highest total concentration for paraben mixtures is 0.8% (w/w) [6]. Consequently, the development of analytical methods of parabens in cosmetic products has practically demanded for consumer health.

Corresponding author. Tel.: +86 551 3607072; fax: +86 551 3603388. E-mail address: yzhe@ustc.edu.cn (Y.-Z. He).

For paraben separation and determination, a large number of chromatographic methods have been reported. High performance liquid chromatography (HPLC) is the most common method used for detecting these compounds, which often combines a pretreatment procedure to remove non-polar matrices and uses a gradient elution to improve the resolution typically [710]. In recent years, capillary electrophoresis (CE) including micellar electrokinetic chromatography (MEKC) was perceived to be an attractive separation technique in paraben analysis due to its high efciency, low reagent consumption and high analysis speed [1115]. In spite of its versatility, some limitations, such as discontinuous sample injection, limited analysis throughput, low sensitivity and large matrix effects, still call for the improvement of CE further. In addition, off-line sample pretreatment is time consuming and often requires considerable skill to maintain good precision. Flow injection analysis (FIA) has already been employed in paraben analysis during past 5 years [1619]. The combination of FIA with CE has shown to be of great value and practical applicability as demonstrated by several papers

0039-9140/$ see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.talanta.2007.09.007

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[2028]. The advantages of FIACE system include outstanding reproducibility in peak response and retention time and enhanced sample throughout. These systems have been applied to on-line sample pretreatment, including sample ltration [26], dialysis [22], gas diffusion [24], ion exchange preconcentration [28], derivation [25] and solid-phase extraction [27], etc. The sample pretreatment prior to CE separation for the determination of parabens in cosmetic products includes liquidliquid extraction, ion exchange clean up, protein precipitation, etc. which are laborious, time consuming and insufcient for the removal of interference substances [29]. Solid-phase extraction (SPE) [30] can avoid or minimize the shortcomings of liquidliquid extraction, particularly, for its large organic solvent consumption. To remove oil matrices in oil-based lotions and creams, and eliminate the interference in the paraben analysis, solid-phase extraction is used as the sample clean up and preconcentration procedure in this paper. Focusing on the interface techniques in the combination of FIA with CE, several designs of on-line interfaces are reported [21,31,32]. In those systems, the sample solutions passed through the reservoir of running buffer, which was able to cause the buffer contamination and increase its consumption for injecting the sample solution and removing the polluted buffer. In this paper, an improved interface is adopted to protect the running buffer from contamination and reduce the buffer consumption, especially, containing expensive reagents, and is used for coupling continuous on-line SPE pretreatment with MEKC separation to determine three parabens in cosmetic products.

2.2. FIASPEMEKC system A 1229-HPCE Analyzer (Institute New Tech. Appl., Beijing, China) detecting at 254 nm and an N-2000 double-channel chromatography processor (Institute Info. Engineering, Zhejiang Univ., Zhejiang, China) were used throughout the MEKC separation. An uncoated fused silica capillary with 50 m i.d., 70 cm total length and 40 cm effective length was purchased from Yongnian Chromatogr. Components Ltd. (Hebei, China). A power supply of negative high voltage was employed for the operation safety of the interface. An XW-80A vortex mixer (Jingke, Shanghai, China) was used for mixing sample solutions. An FIA system (IFIS-C, Ruimai Electr. Tech. Ltd., Xian, China) equipped with two peristaltic pumps, one eight-way injection valve and one 90 L sample loop was employed for the automated FIA operation. 0.5 mm i.d. PTFE tube and 1.2 mm i.d. Viton tubing (Cole-Parmer Instr. Co., USA) was used as the conduits and pump tubing of the FIA system, respectively. A homemade split ow interface of the FIASPEMEKC system is shown in Fig. 1. The schematic diagram of the system is illustrated in Fig. 2. The interface was prepared from a 1 mL plastic pipette, of which the upper part 25 mm and the lower part 3 mm from the pipette tip were cut off and the latter formed a hole of 1.6 mm diameter. To introduce the running buffer solution into the interface, a short silicone tube was connected with the side of the pipette through a 5 mm diameter hole. A Tee connector (TC) was used to joint a 0.5 mm i.d. and 1.5 mm o.d. PTFE tube, a separation capillary (SC) and a sample loop (SL), where the pretreated sample solution was lled and delivered by the pumps. The PTFE tube with a slope end and a grounded Pt electrode were xed on the top of the pipette by a silicone rubber seal (SRS1), and the PTFE tube was inserted into the pipette tip connected with an injection valve (V) by a 1.2 mm i.d. Viton tubing (VT) to expel waste solutions from the interface. Before

2. Experimental 2.1. Reagents and solutions Methyl p-hydroxybenzoate (MP), ethyl p-hydroxybenzoate (EP), propyl p-hydroxybenzoate (PP), butyl p-hydroxybenzoate (BP), sodium tetraborate, sodium dodecyl sulfate (SDS), sodium hydroxide, hydrochloric acid, ethanol, methanol, acetonitrile (ACN), etc. were of analytical grade and obtained from Chemical Reagent Co. (Shanghai, China). Tri-distilled water was used to prepare solutions from an SZ-3 distilled water system (Huxi Anal. Instr. Factory, Shanghai, China). Each paraben stock solution of 2.00 mg/mL was prepared in acetonitrile. The calibration solutions with different concentrations were prepared by diluting the stock solutions with 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 20% acetonitrile (v/v). A 20 mmol/L sodium tetraborate (pH 9.3), the sodium tetraborate solution containing 100 mmol/L SDS and a mixed solution of 40% (v/v), 10 mmol/L sodium tetraborate buffer (pH 9.3) and 60% (v/v) ethanol were used as the carrier solution for FIA, the running buffer in MEKC and the eluent of SPE, respectively. BP did not be detected in the cosmetic samples, it was used as an internal standard (IS) added into each standard and real sample at a concentration of 50 g/mL. All the solutions were degassed for 15 min by an S-2200 ultrasonic bath (120 W, 35 kHz, J&L Ltd., Shanghai, China) before use.

Fig. 1. Schematic diagram of splitow interface: (a) injecting sample solution; (b) aspirating separation buffer. PTFE: polytetrauoroethylene tube; Pt: platinum electrode; SB: separation buffer; SC: separation capillary; S/C: sample/carrier solution; SRS: silicone rubber seal; TC: Tee connector; VT: Viton tubing; SRT: silicone rubber tubing; V: injection valve; W: waste.

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taining expensive reagents, such as chiral selection reagent or low conductivity electrolytes. A homemade SPE microcolumn was prepared from a plastic pipette tip with two ends of 5 mm i.d. and 1.5 mm i.d and packed tightly with 50 mg C8 -bonded silica (50 m particle size, Daisogel, Japan). A small amount of quartz wool was placed at both ends of the microcolumn to avoid loss of the silica particles. Each end of the column was connected with PTFE tubes by Viton tubing. To minimize dispersion, the column was arranged to introduce the sample stream from the narrow end, while to enter the eluent from the wider end [27,33]. 2.3. Sample pretreatment For water-based lotion, 1.00 g lotion was mixed with 2.0 mL, 50 mmol/L sodium tetraborate buffer and 2.0 mL acetonitrile, diluted to 10 mL with water, and then degassed by the ultrasonic bath for 5 min. For oil-based cream, gel and lotion samples, 1.00 g sample was stirred with 5.0 mL ethanol by the vortex mixer for 3 min, ltered through 0.45 m nylon membrane, washed with a mixed solution of 12.5 mL, 20 mmol/L sodium tetraborate buffer and 5.0 mL acetonitrile, diluted to 25.0 mL with water, and then degassed by the ultrasonic bath for 5 min. The analytical recovery was examined by spiking the standard solutions of three parabens into real samples. 2.4. Analytical procedure the sample analysis, the silicone rubber seal (SRS1) with the TC and PTFE tube was removed from the pipette and the running buffer solution was lled into the pipette until the liquid level reached the side hole of the pipette. Then the assembled part was replaced onto the pipette and the PTFE tube should be inserted into the pipette tip about 1.2 mm. With the interface, the running buffer in the pipette did not be polluted by the sample solution passed through, but the electric current was conducted between the running buffer and separation capillary inside the PTFE tube. On the other hand, the running buffer solution was able to be aspirated into the PTFE tube by P2 after injecting the sample solution into the separation capillary electrokinetically. It is benecial to reduce the consumption of running buffer conTable 1 Operation processes of on-line FIASPEMEKC Step Figure Duration (s) Flow rate (mL/min) Pump 1 1 2(a) 60 1.0 Pump 2 1.0 Injection Manifold washing and column equilibrating Sample loading Sample eluting Sample transporting Sample injection Buffer aspirating MEKC analysis Valve positon Function

Fig. 2. Schematic diagram of FIASPEMEKC system: (a) injection position; (b) ll position. P1 and P2: peristaltic pumps; V: injection valve; CL: microcolumn packed with C8 sorbent; SC: separation capillary; HV: negative high voltage; IF: interface; R: buffer reservoir; D: detector; SL: sample loop; BF: blind tting; C: carrier solution; S: sample solution; EQ: equilibrating solution; E: eluent; W: waste.

A daily start-up of FIASPEMEKC was carried out by successively rinsing the separation capillary with 1.0 mol/L HCl (5 min), deioned water (1 min), 1.0 mol/L NaOH (10 min), deioned water (1 min) and the running buffer (10 min) via the capillary outlet by pressure, and washing the SPE column with 80% (v/v) acetonitrile by Pump 1 (P1) at a ow rate of 1.0 mL/min for 3 min. The analytical processes of the FIASPEMEKC system are listed in Table 1. Before sample loading, the SPE column packed with C8 -bonded silica should be equilibrated with the sample medium of 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 20% (v/v) acetonitrile by Pump 1 (P1) at a ow rate

2 3 4 5 6 7
a b c

2(a) 2(b) 2(a) 2(a) 2(b) 2(b)

60 20 10 20 10 420b /540c

0.6 0.75 Off Off Off Off

Off Off 1.0 Off 1.0a Off

Injection Fill Injection Injection Fill Fill

The ow rate of Pump 2 reversed and the separation buffer was aspirated into the PTFE tube of the interface. Without using IS. With BP as IS.

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of 1.0 mL/min for 60 s; meanwhile, the manifold was washed with the carrier solution, as shown in Fig. 2(a). During sample loading, the sample solution was introduced into the SPE column at a ow rate of 0.6 mL/min for 60 s by P1. In elution step, the valve was actuated to ll position, as shown in Fig. 2(b), and the retained analytes were eluted by the eluent of 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 60% (v/v) ethanol at a ow rate of 0.75 mL/min for 20 s. In transporting step, the valve returned to injection position again and the eluted sample solution was transported from the sample loop (SL) to the PTFE tube inside the interface by the carrier stream with a ow rate of 1.0 mL/min for 10 s. With both the pumps stopped, a fraction of sample solution was electrokinetically injected into the separation capillary (SC) by its electroosmotic ow (EOF) at 6 kV for 20 s. Then the valve returned to ll position and Pump 2 (P2) reversed its ow direction to aspirate separation buffer containing SDS, which was supplied from the side hole of the pipette. Although SDS is not an expensive reagent, it is proved that the interface for saving reagent can be performed in this FIASPEMEKC system. Subsequently, the MEKC analysis was carried out at 15 kV for 7 min without using IS or 9 min using IS. The next analysis cycle started from the rst step for washing FIA conduit and equilibrating SPE column again. 3. Results and discussion 3.1. Selection of FIAMEKC conditions For simultaneous determination of MP, EP and PP by FIASPEMEKC, the analytical conditions were investigated in the order of MEKC separation and SPE pretreatment. In this section, the separation conditions were optimized in accordance with the separation parameters, including peak area, migration time, separation efciency and resolution. 3.1.1. Inuence of separation buffer components Four parabens could be separated with 20 mmol/L sodium tetraborate buffer (pH 9.3) by CZE. However, a baseline separation of MP and EP could not be achieved. Then FIAMEKC was investigated for the analyte separation. By examining different sodium tetraborate concentration from 5 to 50 mmol/L (pH 9.3) containing 50 mmol/L SDS, 20 mmol/L sodium tetraborate was used to prepare the separation buffer, because the migration time of the analytes increased and the separation efciency decreased with the sodium tetraborate concentration higher than 20 mmol/L, whereas the resolution decreased with the concentration lower than 20 mmol/L. The inuence of buffer pH was examined from 8.5 to 10.0 with 20 mmol/L sodium tetraborate buffer containing 50 mmol/L SDS. It was found that the pH value of the separation buffer did not inuence the paraben resolution obviously. However, the migration time increased with the pH value because the pKa values of the parabens were around 8.4 and the hydroxy group of parabens dissociated higher than pH 8.4 [13]. By considering the migration time, the pH value of the separation buffer was chosen at 9.3.

Fig. 3. Effect of SDS concentration on resolution of MP, EP, PP and BP. ( ) Resolution of MP and EP, ( ) resolution of EP and PP and ( ) resolution of PP and BP. Standard solution: 100 g/mL MP, EP, PP and 50 g/mL BP in 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 20% (v/v) ACN; running buffer: 20 mmol/L sodium tetraborate buffer (pH 9.3) containing different concentrations of SDS; carrier solution: 20 mmol/L sodium tetraborate buffer (pH 9.3); sample loop volume: 90 L; working voltage: 15 kV; injection time: 20 s; detection wavelength: 254 nm.

SDS concentration had a strong effect on the resolution between MP and EP, which was examined from 20 to 150 mmol/L, as shown in Fig. 3. These two compounds could not be separated on baseline with the concentration lower than 100 mmol/L, and with the concentration higher than 100 mmol/L of the migration time and electric current increased, which could enhance the thermal effect and peak broadening. By compromising the resolution and thermal effect, a nal concentration of 100 mmol/L SDS was chosen. So 20 mmol/L sodium tetraborate running buffer (pH 9.3) containing 100 mmol/L SDS was used in the paraben separation. 3.1.2. Inuence of separation voltage For the separation of the four parabens, the effect of the separation voltage was investigated in the range from 8 to 20 kV. As being expected, the migration time was reduced by increasing the separation voltage. However, the resolution decreased with the separation voltage higher than 15 kV owing to the increase of Joule heating. Moreover, the current increased to 86 A and enhanced baseline noise with the separation voltage of 20 kV. For the concurrent consideration of migration time and resolution, the separation voltage was established at 15 kV in the separation. 3.1.3. Introduction of sample zone into separation capillary To obtain optimal sensitivity and separation efciency by the FIASPEMEKC system, the sample loop volume and transporting ow rate were investigated. For transporting the pretreated sample to the PTFE tube in the interface and injecting into the separation capillary, sample loop volume and transporting ow rate determined the intercepted part of the eluted sample solution and the injecting position of the sample zone, which could inuence the sample amount introduced into the separation capillary. First, the sample loop volume was examined in the range of 30140 L with an eluting ow rate of 0.8 mL/min

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for 20 s. The peak area increased with the sample loop volume, meanwhile, the peak width was broadened and separation efciency was reduced because of the sample zone dispersion. A sample loop of 90 L was selected for compromising the sensitivity and separation efciency. The volume of connecting conduit between the sample loop and interface was about 30 L. The transporting ow rate ranged from 0.6 to 1.4 mL/min with the operating time of 10 s was also investigated in this experiment. It was found that the peak areas of the parabens were reduced with the ow rate lower than 0.8 mL/min or higher than 1.0 mL/min. The transporting ow rate between 0.8 and 1.0 mL/min was the best to inject sample solution into the separation capillary. So a transporting ow rate of 1.0 mL/min was chosen nally. 3.2. Selection of SPE conditions Before separating the parabens, the samples should be pretreated to remove the sample matrices and eliminate the analytical interference. In the proposed method, a pretreatment process of SPE was investigated for selecting an appropriate eluent of SPE elution and MEKC separation, concentrating the parabens and removing the sample matrices. C8 - and C18 bonded silica was investigated as the SPE sorbent. The former was selected because C18 -bonded silica could result in a lot of bubbles during eluting process. With the sorbent of C8 -bonded silica in SPE, we expected that the parabens could be retained on the SPE column and eluted by an eluent containing organic solvent, whereas oil matrices were not retained on the C8 column during sample loading. To achieve the effective pretreatment, the eluents containing three water-soluble solvents were investigated in the experiment, viz., 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 50% (v/v) methanol, ethanol and acetonitrile, respectively. The elution was examined by using a mixed standard solution containing 50 g/mL MP, EP, PP and BP, respectively, which passed through the SPE column at 0.6 ml/min for 60 s and were eluted with different eluent and volume from 50 to 500 L at a ow rate of 0.6 mL/min. The eluted parabens were transported to the interface and separated by MEKC. It was found that the eluent containing ethanol was the best among three eluents for the parabens. The eluting results of the parabens were similar. Taking MP for example, the peak area was 1.2 and 1.7 times of those containing methanol and acetonitrile, respectively. The eluent volume lower than 150 L could reduce the peak area due to the incomplete eluTable 2 Analytical characteristics of FIASPEMEKC MPa LOD (S/N = 3) (g/mL) Peak area R.S.D. (%) Regressive equations Correlative coefcient Linear range (g/mL)
a b

tion, whereas higher than 150 L could result in the analyte dispersion. Therefore, 150 L eluent volume was chosen in this work. With 150 L eluent, the inuence of eluting ow rate on the peak area of three parabens was investigated. The peak areas of the parabens increased with the eluting ow rate from 0.3 to 0.75 mL/min and decreased rapidly higher than 0.75 mL/min. It manifested that different ow rate could inuence the intercepted part of the eluted sample zone and high ow rate lead to an incomplete elution. In accordance with the experimental results mentioned above, 0.75 mL/min was selected as the eluting ow rate in this work. The inuence of different ethanol concentration on the peak area of the parabens was also investigated. When the ethanol concentration increased from 20 to 60% (v/v), the peak area of parabens was enhanced. However, when the concentration was higher than 60%, the peak area kept constant almost and the eluted solution became cloudy. So 20 mmol/L sodium tetraborate buffer (pH 9.3) containing 60% (v/v) ethanol was adopted as the eluent for the parabens in SPE pretreatment. 3.3. Analytical characteristics of FIASPEMEKC The detection limits, reproducibility, regressive equations, correlative coefcients and linear ranges were investigated for the proposed method. The calibration curves and corresponding regressive equations were obtained with ve concentration levels of mixed standard solutions. Each point on the calibration curves corresponded to the mean value obtained from three times of individual measurement. The regressive equations, correlative coefcients and linear ranges for MP, EP and PP with and without using IS are compared in Table 2. The peak area R.S.D. and correlative coefcient using IS are better than those without using IS. The electropherogram of a mixed standard solution containing 50 g/mL of each paraben is shown with three consecutive injections in Fig. 4(a). The R.S.D. (n = 5) of peak area for three parabens was <2.3% using IS and <2.8% without using IS. The day-to-day precision using IS was 3.1, 3.8 and 3.3% for MP, EP and PP, respectively. The analytical period using IS was 12 min by the FIASPEMEKC method. The LODs were calculated with three times R.S.D. of baseline noises (n = 11). The breakthrough volume of the SPE column at a ow of 0.6 mL/min was found to be 2.0 mL with the three parabens solution of 50 g/mL each. By loading the analyte solution at 0.6 mL/min for 3 min, the enrichment factors of 65,

EPa 0.1 2.7 Y = 4.35X + 7.50 0.9997 3500

PPa 0.07 2.8 Y = 9.56X 3.38 0.9974 2500

MPb 0.09 1.7 Y = 0.442X + 0.204 0.9996 2500

EPb 0.1 2.3 Y = 0.911X + 0.073 0.9998 3500

PPb 0.07 2.2 Y = 1.24X 0.130 0.9992 2500

0.1 2.0 Y = 3.20X + 1.34 0.9991 3500

Without using IS.Y, peak area (mV s); X, standard concentration (g/mL). With BP as IS.Y, peak area ratio analyte/IS; X, concentration ratio analyte/IS.

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Table 3 Concentrations of parabens determined in commercial cosmetic samples Sample Parabens Detected concentration (g/mL)a 42 72 125 46 83 189 67 112 31 129 86 28 105 158 Percentage (% w/w)a 0.105 0.180 0.312 0.115 0.208 0.189 0.067 0.280 0.076 0.322 0.215 0.070 0.262 0.395 Recovery (%)a 93.6 92.3 98.6 90.4 94.7 101 93.7 91.3 98.2 95.2 91.6 94.7 90.4 95.3 Detected concentration (g/mL)b 42 74 122 42 85 192 70 111 33 128 85 30 110 150 Percentage (% w/w)b 0.105 0.185 0.305 0.105 0.212 0.192 0.070 0.278 0.082 0.320 0.212 0.075 0.275 0.375 Recovery (%)b 94.4 94.1 99.8 92.7 95.5 98.0 96.9 93.2 101 102 95.8 96.4 92.2 97.8

Cream a Cream b Cream c Water-based lotion a Water-based lotion b Oil-based lotion a Oil-based lotion b Gel a

MP PP PP MP PP PP PP MP PP PP MP EP PP MP

Gel b
a b

Without using IS. With BP as IS.

68 and 72 for MP, EP and PP, can be obtained, respectively, compared with the FIAMEKC without the SPE pretreatment. However, the sample loading time of 1 min was used because the amounts of parebens in cosmetic products could be detected even without the preconcentration. In this work, the SPE pretreatment was used for the clean up of the sample matrices and the enrichment factors of the parabens were only higher than 10.

3.4. Determination of parabens in cosmetic products The proposed FIASPEMEKC system was applied to determine three parabens in commercial cosmetic products, including water-based lotions and oil-based lotions, gels and creams. After off-line preliminary pretreatment described in Section 2.3, the sample solution was loaded on and eluted from the SPE column, and analyzed by MEKC. The peaks were identied by their relative retention time compared with IS. The analytical recovery of the proposed method, including extraction and separation procedure, was based on spiking experiments. The recovery with and without using IS are compared in Table 3, ranging from 92.2 to 102% using IS and 90.4 to 101% without using IS. The concentrations of three parabens found in nine cosmetic products are also listed in Table 3. The electropherograms of an oil-based lotion using BP as IS by three consecutive injections is shown in Fig. 4(b). It shows that the matrix interference was not observed owing to the high selectivity of the SPE pretreatment. The determination results of the three parabens were lower than the maximum addition levels established by China [6]. 4. Conclusion

Fig. 4. Electrochromatograms of three consecutive injections with (a) a mixed standard solution of three parabens and (b) a sample of oil-based lotion using IS by FIASPEMEKC. Peak identication: 1: MP; 2: EP; 3: PP; IS, PP. Standard solution: 50 g/mL MP, EP and PP in 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 20% ACN (v/v); running buffer: 20 mmol/L sodium tetraborate buffer (pH 9.3) containing 100 mmol/L SDS; carrier solution: 20 mmol/L sodium tetraborate buffer (pH 9.3); sample loading: 0.6 mL/min for 60 s; sample elution: 0.75 mL/min for 20 s with 10 mmol/L sodium tetraborate buffer (pH 9.3) containing 60% (v/v) ethanol; sample loop volume: 90 L; working voltage: 15 kV; injection time: 20 s; detection wavelength: 254 nm. Fifty microgram per millilitre of BP was added as IS into the standard and real sample solutions.

The on-line pretreatment and separation method with the FIASPEMEKC system has been used to analyze three parabens of MP, EP and PP successfully. In the combination system of FIASPEMEKC, the modied splitow interface with low reagent consumption was introduced for coupling on-line sample pretreatment with MEKC separation. The throughput of ve samples per hour was achieved for standard and sample solutions with the baseline separation of the parabens. With the proposed method, the analytical sensitivity and selectivity

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were improved in the paraben determination and sample matrix removal. Acknowledgements The authors acknowledge the National Science Foundation of China (nos. 20675075 and 20275035) for nancial support to carry out this work. References
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