Abstract
Objectives
The objective of this study was to develop and validate an automated solid-phase extraction (SPE) coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the detection of sodium pentachlorophenolate (PCP-Na) residues on cutting boards. Given the potential hazards and environmental persistence of PCP-Na, a sensitive and reliable method is crucial for monitoring its residues in food contact materials to ensure consumer safety.
Methods
Wood shavings from cutting boards were extracted using 10% methanol in water, followed by purification using an automated SPE system. The eluent was concentrated, reconstituted, and analyzed by UPLC-MS/MS. An isotope-labeled internal standard was used to mitigate matrix effects, enhancing detection sensitivity. The method was validated by assessing linearity, limit of detection (LOD), limit of quantification (LOQ), recovery rates, and relative standard deviations (RSDs) across various concentration levels.
Results
The method demonstrated excellent linearity over a concentration range of 0 to 100 μg/L with a regression equation of Y = 1.035X−0.7771 and an R² of 0.9996. The LOD and LOQ were determined to be 0.4 and 1.0 μg/kg, respectively. Recovery rates ranged from 71.75% to 96.50% with RSDs between 5.19% and 16.66%. When applied to 30 market cutting board samples, PCP-Na residues were detected in 50% of the samples, with concentrations ranging from 0 to 83,990 µg/kg.
Conclusion
This study presents a robust UPLC-MS/MS method for the detection of PCP-Na on cutting boards, offering improved sensitivity and simplified sample preparation. The high detection rate in commercial samples underscores the need for stringent monitoring and regulatory measures to mitigate the exposure risk to consumers.
Keywords: Sodium pentachlorophenolate, UPLC-MS/MS, solid-phase extraction, food safety, cutting boards, method validation.
Introduction
Sodium pentachlorophenolate (PCP-Na) is a chlorinated aromatic compound and a sodium salt derivative of pentachlorophenol. It has been extensively used as a wood preservative, herbicide, and molluscicide due to its broad-spectrum effectiveness against fungi and insects. However, PCP-Na is known for its persistence in the environment and resistance to biodegradation, raising concerns about its potential for bioaccumulation and long-term environmental and health risks. This compound has historically been employed in various industrial applications to protect wood products from decay and insect damag. 1 The widespread use of PCP-Na has raised serious environmental and health concerns due to its toxicity, persistence, and potential for bioaccumulation. Chronic exposure to even low concentrations of PCP-Na has been associated with carcinogenicity, endocrine disruption, and other severe toxic effects. As a result, regulatory restrictions have been implemented in many countries to minimize its use and prevent further contamination of environmental and food contact materials.2–5 These concerns have prompted regulatory restrictions in many countries, limiting its use to prevent further contamination. 6 Despite these regulations, PCP-Na residues continue to be detected in various environmental matrices, including soil, water, and food contact surfaces, such as cutting boards.7,8 This ongoing presence poses a risk to human health, as chronic exposure to PCP-Na can lead to severe toxic effects, including carcinogenicity and endocrine disruption. 9 Therefore, monitoring PCP-Na residues, particularly on surfaces that come into direct contact with food, is crucial for mitigating these risks and ensuring food safety.
Current analytical techniques for detecting PCP-Na residues, such as gas chromatography-mass spectrometry (GC-MS/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), are widely used for the detection of similar organic pollutants, including pentachlorophenol (PCP) and other environmental contaminants. For instance, GC-MS/MS often requires complex sample preparation steps, such as derivatization, which can introduce variability and reduce sensitivity. Additionally, detecting low concentrations of PCP-Na in complex matrices, like cutting boards, presents a challenge due to potential matrix interference. Additionally, GC-MS/MS may struggle with detecting low concentrations of PCP-Na in complex matrices. In contrast, LC-MS/MS, especially when combined with automated solid-phase extraction (SPE), offers several advantages, including higher sensitivity, specificity, and the ability to detect low concentrations of PCP-Na even in complex matrices. The use of automated SPE not only simplifies sample preparation but also reduces human error and improves reproducibility, making it a reliable and efficient method for routine monitoring of PCP-Na residues. LC-MS/MS has been widely applied in detecting various contaminants in food and environmental samples, demonstrating its effectiveness and robustness. For example, LC-MS/MS has been used for the detection of pharmaceuticals in water samples, providing precise and reliable results. 10 Similarly, it has been employed to monitor pesticide residues in food, ensuring compliance with safety regulations. 11 The use of automated solid-phase extraction (SPE) coupled with LC-MS/MS has further enhanced the detection capabilities by streamlining sample preparation, reducing analysis time, and improving reproducibility. 12 This combination has been validated in various studies, demonstrating its effectiveness in detecting residues in different environmental and food-related matrices. However, further validation is needed to specifically assess its applicability for ensuring the safety of food contact materials.13,14
Given that current LC-MS/MS methods have limitations in detecting PCP-Na residues at very low concentrations, our research aims to develop and validate a more sensitive and reliable LC-MS/MS method tailored for PCP-Na. In addition, previous studies have demonstrated the successful application of high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for the multi-residue screening of pesticides, including in complex food matrices, which highlights the versatility and potential of this approach for analyzing contaminants such as PCP-Na. 15 This study provides a significant advancement in the detection of PCP-Na residues on food contact materials. By offering a more sensitive and streamlined method for monitoring this contaminant, we contribute to safer food handling practices and help mitigate the health risks associated with prolonged exposure to PCP-Na. This method has broad applications in regulatory monitoring and public health research.
Materials and methods
Sample preparation and solid-phase extraction
Cutting board samples were collected by placing aluminum foil underneath the board and drilling holes to collect wood shavings according to the nine-point pattern shown in Figure 1. The wood shavings were collected on the aluminum foil, further ground using a grinder, and stored in polyethylene plastic bags with sample identification labels. The samples were stored at room temperature in a dry environment for future analysis. A minimum sample amount of 10 g was required; if the total weight was less than this, additional sampling points outside the nine-point pattern were randomly added.
Figure 1.
Schematic diagram of the nine-point sampling method used for collecting wood shavings from cutting boards. The sampling follows a regular and reverse plum blossom pattern, as shown in the circular and rectangular layouts. Aluminum foil is placed beneath the board to collect the samples, which are further processed for PCP-Na analysis.
A total of 30 cutting board samples were randomly purchased from various markets in Shenzhen in 2023 for the analysis of sodium pentachlorophenolate (PCP-Na) residues. The sample size of 30 was chosen to capture potential variability in PCP-Na contamination across different vendors and to provide statistically meaningful results. This study was designed as a cross-sectional analysis to evaluate the presence of PCP-Na in food contact materials.
For each sample, 1.0 g of the prepared sample was placed into a centrifuge tube, and 10 mL of 10% methanol in water (1:9) was added. The mixture was vortexed for 30 min and then soaked overnight at 4°C. The mixture was centrifuged at 5000 rpm for 10 min, and 5 mL of the supernatant was collected and spiked with 20 μL of an internal standard solution (PCP-13C, 1.0 mg/L, obtained from Shanghai Anpel Laboratory Technologies Inc.).
The sample was then purified using an automated solid-phase extraction (SPE) system-ASPEC GX-274 automated solid-phase extraction system (Gilson, USA). The SPE column used was a mixed anion exchange solid-phase extraction column (MAX, 3 mL, 60 mg, Waters, USA). The SPE conditions were as follows:
Activation with 4 mL methanol and 4 mL water.
Loading the sample.
Washing with 4 mL water and 4 mL methanol.
Rinsing with 3 mL of 2% formic acid in a 50:50 methanol-water solution.
Drying under vacuum for 10 min.
Elution with 4 mL of 4% formic acid in methanol solution.
The eluent was collected and concentrated to approximately 0.5 mL under a nitrogen stream. The final volume was adjusted to 2 mL with water, mixed thoroughly, and filtered through a 0.22 μm membrane filter before LC-MS/MS analysis.
Liquid chromatography conditions
The liquid chromatography (Shimadzu LC-40A, JP) analysis was performed using a DiKMA Eudeavorsil column (1.8 μm × 2.1 mm × 50 mm). The column temperature was maintained at 40°C. The mobile phase consisted of 5 mmol/L ammonium acetate in water (solvent A) and pure methanol (solvent B). The flow rate was set to 0.4 mL/min with the following gradient elution program:
0 to 0.1 min: 10% B
to 2 min: 10% to 50% B
to 5 min: 50% to 90% B
5 to 8 min: 90% B
8 to 8.1 min: 90% to 10% B
8.1 to 10 min: 10% B
The injection volume for the samples was 10 μL.
Mass spectrometry conditions
The mass spectrometry (ABSciex 6500+ Triple Quadrupole Tandem Mass Spectrometer (Applied Biosystems, USA) analysis was conducted using an electrospray ionization (ESI) source operating in negative ion mode with multiple reaction monitoring (MRM). The specific conditions were as follows:
Ion spray voltage (IS): −4500 V
Ion source temperature (TEM): 450°C
Curtain gas (CUR): 30 psi
Nebulizing gas (GS1): 55 psi
Auxiliary gas (GS2): 55 psi
Collision gas (CAD): 8 psi
Entrance potential (EP): 10 V
Collision cell exit potential (CXP): 10 V
Dwell time: 100 ms
The MRM parameters for sodium pentachlorophenolate and its internal standard are detailed in Table 1.
Table 1.
MRM ion pair parameters for sodium pentachlorophenolate and internal standard.
| Compound Name | Precursor Ion (m/z) | Product Ion (m/z) | Declustering Voltage (V) | Collision Energy (eV) | Retention Time (min) |
|---|---|---|---|---|---|
| (PCP-Na) | 262.7 | 262.7 | −60 | −13 | 5.03 |
| 264.7 | 201.8 | −60 | −43 | 5.03 | |
| 264.7 | 264.7 | −60 | −13 | 5.03 | |
| 266.7 | 266.7 | −60 | −13 | 5.03 | |
| PCP-C13 | 271.0 | 271.0 | −60 | −13 | 5.03 |
| 271.0 | 207.0 | −60 | −43 | 5.03 |
Quantification method
The quantification of sodium pentachlorophenolate (PCP-Na) was performed using the internal standard method. To prepare the working curve, 1.0 g of blank cutting board samples, which were previously confirmed by laboratory testing to be free of PCP-Na, were selected and processed following the sample preparation method described earlier. These blank samples were then spiked with known concentrations of PCP-Na standard solutions to create matrix-matched working standards. The working curve was generated based on these matrix-matched standards to ensure accurate quantification of PCP-Na in the cutting board samples. The blank matrix solution was then spiked with varying concentrations of PCP standard solution to obtain matrix-matched calibration standards with concentrations of 0, 1, 2, 5, 10, 20, 40, and 100 μg/L. Each calibration standard was spiked with 20 μL of the internal standard (PCP-13C).
These calibration standards were analyzed using the LC-MS/MS system. A calibration curve was constructed by plotting the spiked concentration of the standards (x-axis) against the measured concentration (y-axis), ensuring accurate quantification of the analyte in relation to the internal standard. Linear regression analysis was used to determine the relationship between the concentration and the peak area ratio, which was then used for quantifying the PCP-Na in the sample extracts.
The quantification of sodium pentachlorophenolate (PCP-Na) was performed using the internal standard method. To prepare the working curve, 1.0 g of blank cutting board samples which were previously confirmed by laboratory testing to be free of PCP-Na were selected and processed following the sample preparation method described earlier. For samples with PCP-Na concentrations exceeding the upper limit of the calibration curve, an appropriate dilution was performed to bring the concentration within the calibration range before analysis. This ensures accurate quantification of the analyte, and the final concentrations were adjusted accordingly to reflect the dilution factor.
Results
Linearity and detection limits
The experimental results indicated that the method exhibited excellent linearity over the concentration range of 0 to 100 μg/L. The regression equation was y = 1.035*x −0.7771, with a correlation coefficient (R2) of 0.9996, as shown in Figure 2. The limit of detection (LOD) was determined as the concentration corresponding to a signal-to-noise ratio (S/N) of 3, and the limit of quantification (LOQ) was defined as the concentration corresponding to an S/N of 10. Based on a sample weight of 1.0 g and a final volume of 2.0 mL, the LOD of the method was found to be 0.4 μg/kg, and the LOQ was 1.0 μg/kg.
Figure 2.
Calibration curve for sodium pentachlorophenolate.
Recovery and precision
Eighteen cutting board wood shavings samples (1.0 g each) without the target analyte were selected. The spiked samples were prepared at three concentration levels of PCP-Na in the extract (1, 5, and 20 µg/L), which, considering the sample extraction and final concentration, correspond to 4, 20, and 80 µg/kg in the cutting board samples, respectively. The concentration levels of PCP-Na in the extract (µg/L) were converted to concentrations in the cutting board samples (µg/kg) by considering the extraction and concentration steps. Specifically, the final concentration of PCP-Na in the sample is calculated based on the sample weight in the final extract volume. Six replicate samples were prepared for each concentration level. The samples were processed according to the sample preparation method described in section 2.1 and analyzed using the established LC-MS/MS method.
The relative standard deviation (RSD) of the measurements ranged from 5.19% to 16.66%, indicating good precision. The recovery rates ranged from 71.75% to 96.50%, demonstrating satisfactory accuracy. The detailed results are presented in Table 2.
Table 2.
Recovery and precision of the method for sodium pentachlorophenolate.
| Spiked Concentration | Measured Concentration | Average Concentration | Relative Standard Deviation (RSD) | Recovery Rate | |||||
|---|---|---|---|---|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 | 6 | ||||
| 4.00 μg/kg | 4.08 | 3.88 | 3.72 | 4.12 | 3.68 | 3.68 | 3.84 | 5.19% | 96.50% |
| 20.00 μg/kg | 12.04 | 13.56 | 15 | 14.72 | 18.8 | 12.68 | 14.48 | 16.66% | 72.33% |
| 80.00 μg/kg | 59.20 | 52.00 | 54.80 | 61.60 | 56.80 | 60.00 | 557.40 | 6.23% | 71.75% |
The results demonstrate that the method provides reliable recovery and precision for the quantification of sodium pentachlorophenolate in cutting board samples.
Determination of actual samples
The selection of 30 samples is supported by the Central Limit Theorem, which suggests that a sample size of around 30 is typically sufficient for reliable statistical inference and for approximating a normal distribution of sample means. This sample size is commonly used in exploratory studies to balance the need for sufficient data while accounting for resource limitations.
The samples were prepared and analyzed using a validated LC-MS/MS method. The results are summarized in Table 3. The concentration of PCP-Na detected in the 30 samples ranged from ND to 83,990 µg/kg, with PCP-Na being detected in 50% of the samples. Samples reported as ND indicate concentrations below the limit of detection (LOD). These results indicate significant variability in the presence of PCP-Na across the sampled cutting boards, highlighting the need for further investigation into contamination sources and distribution.
Table 3.
Detection of sodium pentachlorophenolate in market cutting board samples.
| Sample ID | Detection Result (µg/kg) |
|---|---|
| 01 | ND |
| 02 | ND |
| 03 | 6150 |
| 04 | 530 |
| 05 | 59,960 |
| 06 | 34,220 |
| 07 | 83,990 |
| 08 | 590 |
| 09 | 110 |
| 10 | ND |
| 11 | 50 |
| 12 | ND |
| 13 | ND |
| 14 | ND |
| 15 | 13,510 |
| 16 | 9360 |
| 17 | ND |
| 18 | 100 |
| 19 | ND |
| 20 | ND |
| 21 | 3400 |
| 22 | ND |
| 23 | ND |
| 24 | 11,610 |
| 25 | 110 |
| 26 | ND |
| 27 | ND |
| 28 | ND |
| 29 | ND |
| 30 | 4790 |
Discussion
This study successfully established an automated solid-phase extraction-UPLC-MS/MS method for the determination of sodium pentachlorophenolate (PCP-Na) in cutting boards. The method involves sample extraction with 10% methanol in water, followed by centrifugation and purification using an automated solid-phase extraction system. The eluent is then concentrated, reconstituted in the initial mobile phase, filtered, and analyzed by UPLC-MS/MS. The use of an isotope-labeled internal standard for quantification effectively reduces matrix effects commonly encountered in liquid chromatography-mass spectrometry, thereby improving detection sensitivity.
The necessity of detecting PCP-Na residues on cutting boards arises from the potential use of this compound in the wood preservation process to prevent decay, mold, and color loss, which might lead to residues on food contact surfaces. In our study, 50% of the cutting board samples purchased from the market were found to contain detectable levels of PCP-Na, with concentrations ranging from ND to 83,990 µg/kg. This high detection rate, along with the wide concentration range, suggests significant variability in contamination levels across different cutting boards. The presence of such high concentrations of PCP-Na raises concerns about potential consumer exposure through food contact surfaces. These findings indicate the need for further investigation into the factors contributing to this variability, such as differences in manufacturing processes, wood preservation techniques, and market sources.
Compared to existing methods, our UPLC-MS/MS method demonstrates a lower limit of quantification (LOQ) and improved recovery rates with narrower RSD ranges. For instance, a recent study by Jia et al. developed a method for detecting PCP-Na and its metabolites in animal-origin foods with LOQs of 1–2 μg/kg, recovery rates between 60.5% and 119.9%, and RSDs ranging from 0.70% to 12.06%. In contrast, our method achieves an LOQ of 1.0 μg/kg, recovery rates between 71.75% and 96.50%, and RSDs ranging from 5.19% to 16.66%, indicating a more precise and reliable quantification process. For instance, a recent study by Jia et al. developed a method for detecting PCP-Na and its metabolites in animal-origin foods with LOQs of 1–2 μg/kg, recovery rates between 60.5% and 119.9%, and RSDs ranging from 0.70% to 12.06%. 8
Further research is needed to understand the migration of PCP-Na from cutting boards to food, which could provide insights into the actual exposure levels for consumers. Such studies could help quantify how much of the detected PCP-Na on cutting boards transfers to food during normal use, thereby refining the assessment of health risks. Given the high detection rate of PCP-Na in commercial cutting boards, coupled with the significant concentration levels observed in some samples, it is crucial for regulatory authorities to enhance surveillance and implement stricter control measures to prevent the unauthorized use of this compound in food contact materials. Moreover, setting lower permissible limits for PCP-Na residues on food contact surfaces could further protect consumers from potential long-term exposure.
These findings highlight the urgent need for both consumers and regulatory bodies to take action in mitigating the risks associated with PCP-Na residues on food contact surfaces. Enhanced monitoring programs and stricter regulatory measures are crucial for preventing the presence of harmful contaminants in consumer products. In addition, more comprehensive research into the migration behavior of PCP-Na, combined with long-term exposure assessments, will be essential for developing more effective strategies to ensure food safety and protect public health. Enhanced monitoring and stricter regulations are essential to ensure food safety and protect public health.
This study has several limitations. First, while the sample size of 30 cutting boards provides statistically meaningful data, the geographic coverage was limited to markets in Shenzhen, and the results may not reflect contamination levels in other regions. Additionally, the study only examined the presence of PCP-Na residues on cutting boards, without investigating the actual migration of PCP-Na to food during normal use, which limits the ability to fully assess consumer exposure. Another limitation is that all samples were collected in 2023, and it is unclear whether PCP-Na contamination levels might vary over time. Lastly, the analysis was based on a specific method (UPLC-MS/MS) using solid-phase extraction, and further validation with alternative methods could enhance the generalizability of the findings.
Conclusions
This study successfully established an automated solid-phase extraction-UPLC-MS/MS method for the detection of sodium pentachlorophenolate (PCP-Na) in cutting boards. The method demonstrated high sensitivity and precision, making it suitable for routine monitoring. PCP-Na residues were detected in 50% of the samples, with concentrations ranging from ND to 83,990 µg/kg, indicating significant variability in contamination levels. These findings highlight potential risks for consumer exposure, emphasizing the need for further research on PCP-Na migration to food. Additionally, stricter regulatory control and enhanced surveillance are crucial to mitigate the risks posed by PCP-Na residues in food contact materials.
Acknowledgements
The authors would like to thank all individuals who contributed to this study.
Footnotes
Author contribution: Shicheng Liao was responsible for drafting the initial version of the manuscript and involved in the acquisition and analysis of data. Wanna Xu, Jie Jiang, and Honghe Liu contributed to discussions, revisions of the manuscript, and interpretation of the data. Zhuoying Zeng provided the overall conceptual framework, supported the research funding, and finalized the overall logic of the article.
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Funding: The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Shenzhen Key Medical Discipline (No. SZXK066); Shenzhen Postdoctoral Research Fund for Staying in Shenzhen; Guangdong Basic and Applied Basic Research Foundation (No. 2022A151511119); Shenzhen Science and Technology Program (No. JCY20220530150402004).
ORCID iD: Zhuoying Zeng https://orcid.org/0000-0001-9727-7994
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