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User Benefits Application News Introduction Foods can contain various ingredients, including preservatives that inhibit microbial growth. In January 2026, the U.S. federal government recommended limiting intake of ultra-processed foods (UPF), identifying 11 target compounds including propylparaben 1) . Around the same time in Japan, there was a reported recall of confections containing methylparaben, which is not approved domestically. Given the circumstances, more rapid and effective quality-control systems are required to ensure product safety. Organic acids, including those employed as acidulants, are typically analyzed using dedicated systems based on ion- exclusion chromatography. However, the application of reversed- phase chromatography enables the simultaneous determination of preservatives and other additives within a single analytical run. This approach enhances analytical throughput and improves workflow efficiency for compounds with diverse physicochemical properties. In this study, an integrated HPLC system (Fig. 1) is utilized for the rapid and simultaneous analysis of food additives, including preservatives, acidulants, and a bittering agent. Preservatives, acidulants, and other compounds with differing polarities can be quickly analyzed using a general-purpose HPLC system, even without using a dedicated instrument. The photodiode array (PDA) detector built into the integrated HPLC system enables acquisition of chromatograms at any desired wavelength from a single analysis. Analytical Strategy for Food Safety Control Using i-Series HPLC—High-Speed Determination of Additives Associated with Ultra-Processed Food Consumption— Natsuki Iwata High Performance Liquid Chromatograph Analysis of Mixed Standard Solution Twelve food-additive compounds were targeted for analysis: methylparaben, ethylparaben, isopropylparaben, propylparaben, isobutylparaben, butylparaben, benzoic acid, sorbic acid, dehydroacetic acid, citric acid, propionic acid, and caffeine. Table 1 shows the analytical conditions and Fig. 2 shows the chromatograms from analyzing the mixed standard solution (with citric acid and propionic acid at 100 mg/L each and the other ten compounds at 1 mg/L each). Fig. 1 Integrated LC System “i-Series” (LC-2080C 3D) Fig. 2 Chromatograms for Mixed Standard Solution 1. Citric acid, 2. Propionic acid, 3. Caffeine, 4. Sorbic acid, 5. Benzoic acid, 6. Methylparaben, 7. Dehydroacetic acid, 8. Ethylparaben, 9. Isopropylparaben, 10. Propylparaben, 11. Isobutylparaben, 12. Butylparaben 1 2 4 5 6 7 8 9 10 11 12 3 210 nm 230 nm 256 nm 260 nm 272 nm 310 nm 0.0 2.5 5.0 7.5 min 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 mV The PDA detector wavelength was set near the maximum absorption wavelength for each target compound. Using a simultaneous analysis method, all 12 target compounds were eluted within 10 minutes.
Application News Repeatability Table 2 shows the repeatability (%RSD) of retention time and peak area values in six repeated analyses of the mixed standard solution (with citric acid and propionic acid at 100 mg/L each and the other ten compounds at 1 mg/L each). Repeatability of both retention time and peak area values was satisfactory for all compounds. Calibration Curves The calibration curves for the 12 target compounds were highly linear, with coefficients of determination (r 2 ) of 0.9999 or greater. Fig. 3 shows the calibration curves of citric acid and sorbic acid. Table 3 shows the concentration ranges of the calibration curves and coefficients of determination for the target compounds. Compound Retention time Peak area Citric acid 0.08 0.08 Propionic acid 0.04 0.34 Caffeine 0.03 0.15 Sorbic acid 0.03 0.08 Benzoic acid 0.02 0.22 Methylparaben 0.02 0.08 Dehydroacetic acid 0.02 0.10 Ethylparaben 0.02 0.11 Isopropylparaben 0.03 0.06 Propylparaben 0.03 0.13 Isobutylparaben 0.03 0.31 Butylparaben 0.03 0.14 Compound Conc. Range (mg/L) r 2 Citric acid 10 to 1000 0.9999 Propionic acid 10 to 1000 0.9999 Caffeine 0.1 to 10 0.9999 Sorbic acid 0.1 to 10 0.9999 Benzoic acid 0.1 to 10 0.9999 Methylparaben 0.1 to 10 0.9999 Dehydroacetic acid 0.1 to 10 0.9999 Ethylparaben 0.1 to 10 0.9999 Isopropylparaben 0.1 to 10 0.9999 Propylparaben 0.1 to 10 0.9999 Isobutylparaben 0.1 to 10 0.9999 Butylparaben 0.1 to 10 0.9999 0 500 0 1.0 2.0 3.0 4.0 Peak area (x100,000) (mg/L) Conc. Citric acid 0.0 5.0 0 2.5 5.0 7.5 Peak area (x100,000) (mg/L) Conc. Sorbic acid Fig. 3 Calibration Curves (Citric Acid and Sorbic Acid) Table 2 Repeatability (%RSD) in Six Repeated Analyses Table 3 Concentration Ranges of Calibration Curves and Coefficients of Determination (r 2 ) System: i-Series LC-2080C 3D Column: Shim-pack Scepter C18-120 *1 (100 mm × 3.0 mm I.D., 1.9 µm) Flowrate: 0.8 mL/min Mobile Phase: A) 20 mmol/L (sodium) phosphate buffer (pH 2.4) B) Acetonitrile Time Program: 2 % B (0 min) → 25 % B (2.50 min) → 32 % B (4.50 min) → 60 % B (9.00 - 10.00 min) → 2 % B (10.01 - 15.00 min) Column Temp.: 30 °C Injection Volume: 5 µL Vial: Shim-vial H glass *2 Detection: Ch 1: 210 nm, Ch 2: 230 nm, Ch 3: 256 nm, Ch 4: 260 nm, Ch 5: 272 nm, Ch 6: 310 nm *1 P/N: 227-31013-03, *2 P/N: 227-34500-01 Table 1 Analytical Conditions Analysis of Energy Drink Chromatograms of a commercial energy drink are shown in Fig. 4. The sample was diluted 100-fold with ultrapure water and filtered through a 0.2 µm membrane filter before HPLC analysis. Citric acid, caffeine, benzoic acid and ethylparaben were detected from the energy drink. Table 4 (1) shows the concentrations of each compound in the energy drink. Note that the concentrations shown in Table 4 are average concentrations obtained from six repeated analyses of samples after pretreatment.
Application News www.shimadzu.com/an/ Shimadzu Corporation © Shimadzu Corporation, 2026 For Research Use Only. Not for use in diagnostic procedures. This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. See https://www.shimadzu.com/about/trademarks/index.html for details. Third party trademarks and trade names may be used in this publication to refer to either the entities or their products/services, whether or not they are used with trademark symbol “TM” or “”. Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice. First Edition: Jun. 2026 01-01163-EN Shim-pack Scepter and shim-vial are trademarks of Shimadzu Corporation or its affiliated companies in Japan and/or other countries. References 1) Dietary Guidelines for Americans, 2025–2030 (https://cdn.realfood.gov/DGA.pdf ) 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min 0 10 20 30 40 50 60 70 80 mAU ■Peak 4. Sorbic acid 4 260 nm Compound (1) Energy drink (2) Wine Conc. (mg/L) %RSD Conc. (mg/L) %RSD Citric acid 25.5 0.61 N.D. Caffeine 4.89 0.17 N.D. Sorbic acid N.D. 1.50 0.07 Benzoic acid 5.31 0.11 N.D. Ethylparaben 0.56 0.18 N.D. Analysis of Wine A chromatogram of a commercial wine is shown in Fig. 5. The sample was diluted 100-fold with ultrapure water and filtered through a 0.2 µm membrane filter before HPLC analysis. Sorbic acid was detected from the wine. Table 4 (2) shows the concentrations of each compound in the wine. Note that the concentrations shown in Table 4 are average concentrations obtained from six repeated analyses of samples after pretreatment. Conclusion A rapid reversed-phase chromatographic method for the analysis of multiple food additives was developed using the i- Series LC-2080C 3D. While organic acids such as citric acid are conventionally determined using dedicated ion-exclusion chromatography systems, the proposed approach enables their simultaneous analysis together with preservatives and a bittering agent within a single run. By consolidating the analysis of compounds with diverse physicochemical properties, the i- Series system significantly enhances analytical throughput and minimizes overall analysis time, thereby providing an efficient and streamlined solution for high-speed quality control workflows. Fig. 5 Chromatogram of wine Table 4 Analysis Results (n = 6) 210 nm 272 nm 1 5 8 ■Peak 1. Citric acid ■Peak 3. Caffeine ■Peak 5. Benzoic acid ■Peak 8. Ethylparaben 230 nm 256 nm 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min 0 25 50 75 100 125 150 mAU 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min 0 10 20 30 40 50 60 70 80 mAU 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min 0 10 20 30 40 50 60 70 mAU 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min -5 0 5 10 15 20 25 30 35 40 45 mAU Fig. 4 Chromatograms of Energy Drink 3
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