Journal of Chromatography A, 1146 (2007) 6166

Analysis of abietic acid and dehydroabietic acid in food samples by in-tube solid-phase microextraction coupled with liquid chromatographymass spectrometry
K. Mitani, M. Fujioka, A. Uchida, H. Kataoka
School of Pharmacy, Shujitsu University, Nishigawara, Okayama 703-8516, Japan Received 30 November 2006; received in revised form 20 January 2007; accepted 25 January 2007 Available online 5 February 2007

Abstract A simple and sensitive method for the determination of abietic acid and dehydroabietic acid in food samples was developed using a fully automated method consisting of in-tube solid-phase microextraction (SPME) coupled with liquid chromatographymass spectrometry (LC/MS). These compounds were separated within 5 min by HPLC using an ODS-3 column and 5 mM ammonium formate/acetonitrile (10/90, v/v). Electrospray ionization conditions in the negative ion mode were optimized for MS detection of abietic acid and dehydroabietic acid. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 L of sample using a Supel Q PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. Using the in-tube SPME LC/MS method, good linearity of the calibration curve (r > 0.9998) was obtained in the concentration range from 0 to 50 ng/mL, and the detection limits (S/N = 3) of abietic acid and dehydroabietic acid were 2.9 and 2.1 pg/mL, respectively. The in-tube SPME method showed above 75-fold greater sensitivity than the direct injection method (5 L injection). This method was applied successfully to analysis of food samples without interference peaks. The recoveries of abietic acid and dehydroabietic acid spiked into liquid samples were above 79%, and the relative standard deviations were below 6.6%. These compounds were detected at ng/mL or ng/g levels in various liquid or solid food samples contacted with paper. 2007 Elsevier B.V. All rights reserved.
Keywords: In-tube solid-phase microextraction; Automated sample preparation; Liquid chromatographymass spectrometry; Abietic acid; Dehydroabietic acid; Food samples; Paper container

1. Introduction Abietic acid and dehydroabietic acid are tricyclic diterpene carboxylic acids (Fig. 1), and are the main components of colophony (rosin). These compounds are commonly used as uxing agents in solder, paper sizing agent to make paper more water-resistant, and in printing inks, adhesives and plasticizers. Abietic acid and dehydroabietic acid are well-known pulmonary and skin sensitizers and cause occupational asthma and contact allergy [13]. There have been few cytotoxicological studies of these compounds, although dehydroabietic acid has been reported to increase cell death in human epithelia and broblast cells [4], and to exert toxic effects on human erythrocytes [5] and on polymorphonuclear leukocytes [6]. Abietic acid has

Corresponding author. Tel.: +81 86 271 8342; fax: +81 86 271 8342. E-mail address: hkataoka@shujitsu.ac.jp (H. Kataoka).

been reported to cause lysis of human alveolar epithelial cells [7]. Recently, these compounds have also been reported to exhibit DNA-damaging activity in the rec-assay [8]. Abietic acid and dehydroabietic acid were detected in airborne [2,9] and aquatic environments [1012], and are the main toxicants in pulp and paper mill efuent [13]. These compounds were also found in many consumer products, such as medicaments, cosmetics, and industrial materials [14,15]. Moreover, these compounds were detected in paper and board food packaging [8], and reported about migration from these food container and packaging materials into food-simulating solvents and Tenax TA [9]. However, it has not been investigated whether abietic acid and dehydroabietic acid in the food supply can cause adverse biological effects in humans. Therefore, a sensitive, selective, and simple method to determine the presence and contents of abietic acid and dehydroabietic acid in food samples is required. Analyses of abietic acid and dehydroabietic acid have been carried out mainly by gas chromatography (GC) [1,11],

0021-9673/$ see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.chroma.2007.01.118

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K. Mitani et al. / J. Chromatogr. A 1146 (2007) 6166

Fig. 1. Chemical structures of abietic acid and dehydroabietic acid.

GC/mass spectrometry (GC/MS) [2,3,8], high performance liquid chromatography (HPLC) [10,11,15,16], and LC/MS [8,9]. GC and GC/MS methods require derivatization of these compounds prior to analysis, and HPLC methods with UV and uorometric detection are less sensitive. Most of the above methods require sample preparation steps, such as extraction, concentration, fractionation and isolation, because the analysis of these compounds is complicated by the sample matrix. Solvent extraction under reux, liquidliquid extraction, and solid-phase extraction have been used as sample preparation techniques. However, most of these techniques are complicated and time-consuming. Complicated pretreatment methods may introduce errors, and the use of large volumes of organic solvent poses a health hazard to those performing the analyses and contributes to environmental pollution. Therefore, it is important to develop an efcient sample pretreatment method, and automation will reduce labor and cost. A routine analysis method will also facilitate the processing of large numbers of samples. In-tube solid-phase microextraction (SPME) [17], using an open tubular fused-silica capillary with an inner surface coating as the SPME device, is simple and can be easily coupled on-line with HPLC and LC/MS. In-tube SPME allows convenient automation of the extraction process, which not only reduces the analysis time, but also provides better accuracy, precision, and sensitivity than manual off-line techniques. We have developed an in-tube SPME method for determination of various compounds, such as drugs and environmental contaminants, by coupling with HPLC [1821], LC/MS [2225], and LC/MS/MS [26,27]. The details of the in-tube SPME technique and its applications have been summarized in a number of reviews [2833]. Here, we report a fully automated on-line in-tube SPME LC/MS method for determination of abietic acid and dehydroabietic acid in food samples. 2. Experimental 2.1. Materials Abietic acid and dehydroabietic acid were purchased from Alfa Aesar (Word Hill, MA, USA) and Wako Pure Chemicals (Tokyo, Japan), respectively. Each compound was dissolved in methanol to make a stock solution at a concentration of 1 mg/mL. The solutions were stored at 4 C and diluted to the required concentrations with pure water prior to use. HPLC grade acetonitrile

and water used as mobile phases were purchased from Nacalai Tesque (Kyoto, Japan). All other chemicals were of analytical grade. 2.2. Instrument and analytical conditions The LC/MS system was a Model 1100 series LC coupled with an atmospheric pressure (AP) electrospray ionization (ESI) MS (Agilent Technologies, Boeblingen, Germany). An ODS-3 column (100 mm 2.0 mm, particle size of 5 m; GL Science Inc., Tokyo, Japan) was used for LC separation under the following conditions: column temperature, 40 C; mobile phase, 5 mM ammonium formate/acetonitrile (10/90, v/v); and ow rate, program from 0.2 to 0.35 mL/min for 6 min run. ESI-MS conditions were as follows: nebulizer gas N2 (35 psi); drying gas, N2 (12 L/min, 350 C); fragmentor voltage, 130 V; capillary voltage, 4000 V; ionization mode, negative mode; mass scan range, 100350 amu, 0.94 s/cycle; selected ion monitoring (SIM), m/z 301 (abietic acid) and 299 (dehydroabietic acid); and dwell-times for the ions in SIM, 289 ms. LC/MS data were processed with an HP ChemStation. 2.3. In-tube solid-phase microextraction As shown in Fig. 2, a GC capillary column (60 cm 0.32 mm i.d.) was used as the in-tube SPME device, and placed between the injection loop and injection needle of the autosampler. The injection loop was retained in the system to avoid fouling of the metering pump. Capillary connections were facilitated by use of a 2.5-cm sleeve of 1/16-in. polyetheretherketone (PEEK) tubing at each end of the capillary (1 in. = 2.54 cm). PEEK tubing with an internal diameter of 330 m was suitable to accommodate the capillary used. Standard 1/16-in. stainless steel nuts, ferrules, and connectors were used to complete the connections. CP-Sil 5CB, CP-Sil 19CB, CP-Wax 52CB (Varian Inc., Lake Forest, CA, USA), Carboxen 1010 PLOT and Supel Q PLOT (Supelco, Bellefonte, PA, USA) were tested for comparison of extraction efciency. The autosampler software was programmed to control the in-tube SPME extraction, desorption, and injection. Vials (2 mL) were lled with 1 mL of sample for extraction, and set into the autosampler programmed to control SPME extraction and desorption. In addition, 2-mL autosampler vials with a septum, one containing 1.5 mL of methanol and another containing 1.5 mL of water, were set into the autosampler.

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63

Fig. 2. Schematic diagram of the on-line in-tube SPME LC/MS system. (A) Load position (extraction) and (B) injection position (desorption). (From ref. [26], reproduced with minor modication).

The capillary column was washed and conditioned by two repeated draw/eject cycles (40 L each) of these solvents, and then a 50- L air plug was drawn prior to the extraction step. The extraction of ve estrogens onto the capillary coating was performed by 20 repeated draw/eject cycles of 40 L of sample at a ow rate of 150 L/min with the six-port valve in the LOAD position (Fig. 2A). After washing the tip of the injection needle by one draw/eject cycle of 2 L of methanol, the extracted compounds were desorbed from the capillary coating and transported to the LC column by switching the six-port valve to the INJECT position (Fig. 2B), and detected by the MS system. During the analysis, the SPME capillary was washed and conditioned with mobile phase for the next extraction. 2.4. Preparation of food samples Food samples were purchased from a local supermarket. All samples were stored in their original packaging under recommended conditions (either refrigerated or at room temperature) until use. Liquid samples were used directly after ltration (syringe microlter, 0.45 m, Gelman Science), if necessary. Semi-solid and solid samples were dissolved in aliquots of hot water. The mixtures were centrifuged at 3000 g for 10 min, and the supernatants used for the following in-tube SPME/LCMS analysis. Aliquots of these samples were pipetted into 2-mL screw-cap autosampler vials equipped with silicon/PTFE septa, and 0.1 mL of methanol was added. After the total volume was made up to 1 mL with distilled water, the vials were set onto the sample tray in the autosampler. 3. Results and discussion 3.1. LC/MS analysis of abietic acid and dehydroabietic acid For MS operation, AP and ESI were evaluated for determination of abietic acid and dehydroabietic acid in both positive

and negative ion modes. The ESI negative ion mode was most effective for ionization of these compounds. To select the monitoring ion for each compound, ESI mass spectra were initially analyzed by LC/MS with direct liquid injection into the column. As shown in Fig. 3, each compound gave a very simple spectrum in scan mode for the mass range m/z 100350. Both compounds gave [MH] ions as base ions. The optimum fragmentor voltage was 130 V. Below this level [MH] ion areas decreased. LC separation of abietic acid and dehydroabietic acid was performed using an ODS-3 column. As shown in Fig. 4, these compounds were well separated using 5 mM ammonium formate/acetonitrile (10/90, v/v) as the mobile phase, at linear increase of ow rate from 0.2 to 0.35 mL/min for 6 min. Each compound could be detected selectively by SIM. 3.2. Optimization of in-tube solid-phase microextraction and desorption To optimize the extraction of abietic acid and dehydroabietic acid by in-tube SPME, several parameters, such as the stationary phase of the in-tube SPME capillary column and number and

Fig. 3. ESI-spectra of abietic acid and dehydroabietic acid obtained by direct injection.

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K. Mitani et al. / J. Chromatogr. A 1146 (2007) 6166

Fig. 6. Effects of draw/eject cycles on the in-tube SPME of abietic acid and dehydroabietic acid. Each compound was extracted by draw/eject cycles of 40 L of standard solution (50 ng/mL of abietic acid and 50 ng/mL dehydroabietic acid) using a Supel Q PLOT capillary.

Fig. 4. Typical total ion and selected ion chromatograms obtained from standard abietic acid (50 ng/mL) and dehydroabietic acid (50 ng/mL) by direct injection and in-tube SPME LC/MS in negative ion mode. Peaks: (1) dehydroabietic acid and (2) abietic acid. LC/MS conditions: see Section 2.

volume of draw/eject cycles, were investigated. Five different capillary columns (CP-Sil 5CB, CP-Sil 19CB, CP-Wax 52CB, Carboxen 1010 PLOT, and Supel Q PLOT) were evaluated for extraction efciency. As shown in Fig. 5, the extraction efciency of the porous polymer-type capillary column was better than those of the other columns. As the Supel Q PLOT column has a large adsorption surface area, the extracted amount was greater than that obtained with liquid-phase type columns. The sample load and the amounts of compounds extracted increased with increasing internal diameter and lm thickness of the column.

In in-tube SPME, the extraction time, and ow rate are related to the amounts of compounds extracted. To monitor the extraction time proles of abietic acid and dehydroabietic acid by in-tube SPME, the number of draw/eject cycles was varied from 0 to 25 using a Supel Q PLOT capillary column. As shown in Fig. 6, the extraction reached almost equilibrium after 20 draw/eject cycles of 40 L of sample at a draw/eject rate of 150 L/min. Addition of 10% methanol was effective for extraction. The absolute amounts of abietic acid and dehydroabietic acid extracted by the SPME capillary column were calculated by comparing peak area counts with the corresponding direct injection of the sample solution onto the LC column. At sample concentrations of 100 ng/mL, 36.2 ng (36.2%) of abietic acid and 47.1 ng (47.1%) of dehydroabietic acid were extracted onto the Supel Q PLOT column by in-tube SPME. Although the extraction yields of these compounds were relatively low, reproducibility was good due to the use of an autosampler. The mobile phase was found to be suitable for desorption of abietic acid and dehydroabietic acid extracted into the stationary phase of a capillary column. Dynamic desorption of these compounds from the capillary was readily achieved by switching the six-port valve (Fig. 2B). The desorbed abietic acid and dehydroabietic acid were transported to the LC column by ow of the mobile phase. No carryover was observed because the capillary column was washed and conditioned by draw/eject cycles of methanol and mobile-phase prior to extraction. The extraction and desorption of abietic acid and dehydroabietic acid by the in-tube SPME method were accomplished automatically within 25 min, and automated analysis of about 56 samples per day was possible by overnight operation. 3.3. Limits of detection and calibration curves Abietic acid and dehydroabietic acid provided excellent responses in SIM and the detection limits to give signal-tonoise ratios of 3 under our LC/MS conditions were 2.9 and 2.1 pg/mL (Table 1), respectively. The in-tube SPME method showed about 85 and 75-fold higher sensitivity for each compound than the direct injection method (5 L), because these

Fig. 5. Effects of capillary coatings on the in-tube SPME of abietic acid and dehydroabietic acid. Each compound was extracted by 20 draw/eject cycles of 40 L of standard solution (50 ng/mL of abietic acid and 50 ng/mL dehydroabietic acid) at a ow rate of 150 L/min.

K. Mitani et al. / J. Chromatogr. A 1146 (2007) 6166 Table 1 Linear regression data, detection limits and reproducibilities of abietic acid and dehydroabietic acid by in-tube SPME LC/MS Compound Range (ng/mL) Regression linea Slope (a) Abietic acid Dehydroabietic acid
a b

65

Intercept (b) 3499 7062

Correlation coefcient 0.9999 0.9998

Detection limits (pg/mL) Direct injection (5 L) 248 153 In-tube SPME 2.9 2.1

Intra-day RSD 4.5 5.9 (%)b

Inter-day RSD (%)b 9.9 8.3

0.550 0.550

11,842 16,808

y = ax + b; y is the peak area count and x is the concentration (ng/mL) of each compound. (n = 21). Standard solution containing 10 ng/mL of abietic acid and 10 ng/mL dehydroabietic acid was analyzed. RSD: relative standard deviation (n = 5).

Table 2 Recoveries of abietic acid and dehydroabietic acid spiked into green tea Compound Abietic acid Dehydroabietic acid
a

Spiked (ng/mL) 0.5 0.5

Recovery (%) mean SDa 93.8 6.2 86.9 4.5

Spiked (ng/mL) 5 5

Recovery (%) mean SDa 93.1 2.3 79.3 2.1

SD: standard deviation (n = 3).

compounds in the sample solution were concentrated in the capillary column during draw/eject cycles. To test the linearity of the calibration curve, various concentrations of abietic acid and dehydroabietic acid ranging from 0 to 50 ng/mL were analyzed. Calibration curves were constructed from the peak area counts. As shown in Table 1, a linear relationship was obtained for each compound in this range (seven-point calibration). The correlation coefcients of abietic acid and dehydroabietic acid were 0.9999 and 0.9998, respectively. The within-day and betweenday relative standard deviations (RSDs) were in the range of 4.55.9% and 8.39.9% (n = 5). 3.4. Application to the analysis of food samples Using the in-tube SPME LC/MS method, the following food samples were analyzed: glass bottled drinks, canned drinks, foods packed in plastic containers, and foods packed in paper containers. Liquid samples could be directly analyzed by dilution of the sample without any other pretreatment. Semi-solid

and solid samples were analyzed after extraction with boiling water. Quantication limits of abietic acid and dehydroabietic acid in food samples were 30 and 20 pg/mL, respectively. To conrm the validity of this method, known amounts of these compounds were spiked into green tea, and their recoveries were calculated. As shown in Table 2, the overall recoveries of these compounds were above 79%, and the relative standard deviations were below 6.6%. Fig. 7 shows typical chromatograms obtained from food samples. Abietic acid and dehydroabietic acid were detected without interference peaks by SIM mode detection. As shown in Table 3, abietic acid and dehydroabietic acid were detected in all samples packed in paper containers. However, these compounds were not detected from several food samples packaged in glass bottles, aluminum cans, steel cans or plastic containers, except soft drinks C. In particular, high concentrations of abietic acid and dehydroabietic acid were detected in wheat our and stick sugar, but their concentrations in tea, soft drinks and sake were low. The paper containers used in contact with these foods are laminated with polyethylene lm

Fig. 7. Total ion and selected ion chromatograms obtained from food samples by in-tube SPME LC/MS. (A) Sugar in paper packaging: 3 g of sample dissolved in 100 mL of water; (B) green tea in paper container: sample was used directly for analysis. Peaks: (1) dehydroabietic acid and (2) abietic acid. LC/MS conditions: see Section 2.

66 Table 3 Contents of abietic acid and dehydroabietic acid Sample Package material

K. Mitani et al. / J. Chromatogr. A 1146 (2007) 6166

Contenta /mean SD (n = 3) Abietic acid Dehydroabietic acid ND ND ND 51.2 3.6 100 10 ND ND 503 39 3.4 0.1 ND 1.13 0.09 6.76 0.06 ND 10.0 0.5 62.6 2.8 64.7 4.9 ND ND 0.32 0.01 4.0 0.1 43.8 0.3 10.4 0.1

acid from paper containers into foods was proved. The method described here will be a useful tool for pollution monitoring and determination of abietic acid and dehydroabietic acid in food samples. Acknowledgments This work was supported by grants from The Japan Food Chemical Research Foundation, Japanese Society for Food Science and Technology, and Japan Food Industrial Center, Grants-in-Aid for Basic Scientic Research (B (2), No. 14370279), and Grants-in-Aid for Exploratory Research (No. 16659014). References
[1] S. Sadhra, I.S. Foulds, C.N. Gray, Brit. J. Dermatol. 134 (1996) 662. [2] P.A. Smith, D.R. Gardner, D.B. Drown, G. Downs, W.W. Jederberg, K. Still, Am. Ind. Hyg. Assoc. J. 58 (1997) 868. [3] K. Eriksson, L. Wiklund, C. Larsson, Ann. Occup. Hyg. 48 (2004) 267. [4] T.A. S derberg, A. Johansson, R. Toxicol. 107 (1996) 99. o [5] D.M. Toivola, B. Isomaa, Chem. Biol. Interact. 79 (1991) 65. [6] B. Sunzel, T.A. S derberg, C.O. Reuterving, G. Hallmans, S.E. Holm, L. o H nsr m, Biol. Trace Elem. Res. 31 (1991) 33. a o [7] G.H. Ayars, L.C. Altman, C.E. Frazier, E.Y. Chi, J. Allergy Clin. Immunol. 83 (1989) 610. [8] A. Ozaki, Y. Yamaguchi, T. Fujita, K. Kuroda, G. Endo, Food Addit. Contam. 22 (2005) 1053. [9] A. Ozaki, T. Ooshima, Y. Mori, Food Addit. Contam. 23 (2006) 854. [10] P.A. Smith, P.S. Son, P.M. Callaghan, W.W. Jederberg, K. Kuhlmann, K.R. Still, Toxicology 111 (1996) 225. [11] J.K. Volkman, D.G. Holdsworth, D.E. Richardson, J. Chromatogr. 643 (1993) 209. [12] J.A. Zender, T.R. Stuthridge, A.G. Langdon, A.L. Wilkins, K.L. Mackie, P.N. McFarlane, Water Sci. Technol. 29 (1994) 105. [13] S.N. Liss, P.A. Bicho, J.N. Saddler, Can. J. Microbiol. 75 (1997) 599. [14] Y. Kamaya, N. Tokita, K. Suzuki, Ecotoxicol. Environ. Saf. 61 (2005) 83. [15] B.L. Lee, D. Koh, H.Y. Ong, C.N. Ong, J. Chromatogr. A 763 (1997) 221. [16] K. Hrobo ov , J. Lehotay, I. Ska ani, J. Liq. Chromatogr. Related Technol. n a c 28 (2005) 1725. [17] R. Eisert, J. Pawliszyn, Anal. Chem. 69 (1997) 3140. [18] H. Kataoka, M. Ise, S. Narimatsu, J. Sep. Sci. 25 (2002) 77. [19] K. Mitani, S. Narimatsu, H. Kataoka, J. Chromatogr. A 986 (2003) 169. [20] K. Mitani, S. Narimatsu, F. Izushi, H. Kataoka, J. Pharm. Biomed. Anal. 32 (2003) 469. [21] K. Mitani, F. Izushi, H. Kataoka, J. Anal. Toxicol. 28 (2004) 575. [22] H. Kataoka, H.L. Lord, J. Pawliszyn, J. Chromatogr. Biomed. Sci. Appl. 731 (1999) 353. [23] H. Kataoka, H.L. Lord, J. Pawliszyn, Anal. Chem. 71 (1999) 4237. [24] H. Kataoka, J. Pawliszyn, Chromatographia 50 (1999) 532. [25] H. Kataoka, H.L. Lord, J. Pawliszyn, J. Anal. Toxicol. 24 (2000) 257. [26] K. Mitani, M. Fujioka, H. Kataoka, J. Chromatogr. A 1081 (2005) 218. [27] K. Mitani, H. Kataoka, Anal. Chim. Acta 562 (2006) 16. [28] H. Kataoka, H.L. Lord, J. Pawliszyn, J. Chromatogr. A 880 (2000) 35. [29] H.L. Lord, J. Pawliszyn, J. Chromatogr. A 885 (2000) 153. [30] H.L. Lord, J. Pawliszyn, J. Chromatogr. A 902 (2000) 17. [31] H. Kataoka, Anal. Bioanal. Chem. 373 (2002) 31. [32] H. Kataoka, Trends Anal. Chem. 22 (2003) 232. [33] H. Kataoka, Curr. Pharm. Anal. 1 (2005) 65.

Wine Cup noodle soup Sugar A Sugar B Sugar C Mushroom (broth) Mushroom (broth) Wheat our Black tea Green tea A Green tea B Green tea C Soft drinks A Soft drinks B Soft drinks C Soft drinks C Beer Sake A Sake B Sake C Paper cupc Coffee lterc
a b c

Glass Polystyrene Polyethylene Paper Paper Polyethylene Steel Paper Paper PET Paper Paper PET Paper PET Steel Aluminum Glass Paper Paper Paper Paper

NDb ND ND 262 4 443 12 ND ND 935 86 3.2 0.1 ND 0.58 0.04 0.72 0.04 ND 15.3 2.4 933 33 625 37 ND ND 1.21 0.07 14.1 1.1 75.4 2.9 40.7 0.9

Liquid sample (ng/mL); semi-solid and solid samples (ng/g). Not detectable. Eluate from the sample into 100 mL of boiling water.

to prevent moisture, and their lm thicknesses are generally thicker in liquid foods than solid foods. Therefore, the difference of lm thickness may effect the contents of abietic acid and dehydroabietic acid in foods. On the other hand, remarkably high concentrations of these compounds were detected in steel can and PET bottle in the soft drinks C. It is considered to be polluted from paper, because raw materials powder of the soft drinks was kept in paper container before lling into the steel can and PET bottle as solution. Moreover, these compounds migrated easily from paper cups (laminated with polyethylene lm) and coffee lter paper (100% virgin pulp) into water. Abietic acid and dehydroabietic acid were included 280 and 750 g/g in paper cup, and 15 and 25 g/g in coffee lter, respectively. These results indicate that contamination by these compounds is present in various foods in contact with paper. 4. Conclusions The on-line in-tube SPME LC/MS method developed in the present study can continuously extract abietic acid and dehydroabietic acid from aqueous samples with no requirements for any other pretreatments, which can then be analyzed by LC/MS. This method is automatic, simple, rapid, selective and sensitive, and can be applied easily to the analysis of food samples. Using this method, the migration of abietic acid and dehydroabietic