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The comparison of flow rate calibration methods for high-performance liquid chromatography (HPLC) pump

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Abstract

High-performance liquid chromatography (HPLC) pump has a main function to deliver a mobile phase which needs accuracy and stable flow rate. Recently, the reference of General European OMCL Network 2005: Qualification of Equipment Annex 1 qualification of liquid chromatography equipment presents three flow rate calibration methods that have been used to calibrate the HPLC pump performance such as using standard flow meter, gravimetric method, and volumetric method. In this reference, the calibration methods have not been clearly described. Therefore, this study aimed to investigate and compare the flow rate calibration by the using standard flow meter, gravimetric method, and volumetric method. The flow rate was calibrated at 0.5 and 5 mL/min with three replications. The stability between days of each method was observed for three days. The calibration procedure, traceability chart, and uncertainty budget of each method had been described. The flow rate results were analyzed by using the En ratio. The results showed that the stability of all methods had no statistically significant difference between days when compared to the first day itself. The average measurement flow rates by using standard flow meter, gravimetric method, and volumetric method at 0.5 mL/min were 0.4908, 0.4992, and 0.5012 mL/min, respectively, and at 5 mL/min, they were 4.8346, 4.8272, and 4.8394 mL/min, respectively. These flow rates showed consistent results between each other as confirmed with the En ratio ≤ 1. The precision of the three methods was 0.01 to 0.08 %RSD. This study concluded that the flow rate results of three calibration methods had been consistent with each other. All methods had a good stability and precision for the flow rate calibration of the HPLC pump.

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Acknowledgments

This research has received funding support from the NSRF vi the Program Management Unit for Human Resources and Institutional Development, Research, and Innovation (PMU-B) [grant number B01F640054], and under MOU No. MOU-CO-2564-16159-TH between National Science and Technology Development Agency (NSTDA) and National Institute of Metrology Thailand (NIMT), cooperated with Inctech metrological center co., ltd. The author would like to thank Dr. Cheerapa Boonyakong and Ms. Pornhatai Kankaew from organic analysis group, department of chemical metrology and biometry, National Institute of Metrology (Thailand), for the HPLC technical support.

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Authors and Affiliations

  1. Medical Metrology Laboratory, National Institute of Metrology (Thailand), Pathum Thani, 12120, Thailand

    Prangtip Uthaiwat & Tassanai Sanponpute

  2. Department of Industrial Physics and Medical Instrumentation, King Mongkut’s University of Technology North, Bangkok, 10800, Thailand

    Theera Leeudomwong

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  1. Prangtip Uthaiwat
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  2. Theera Leeudomwong
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  3. Tassanai Sanponpute
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Contributions

PU wrote the main manuscript and processed the laboratory experiments. TS contributed to the experimental design. TS and TL made critical corrections. All authors reviewed the manuscript.

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Correspondence to Tassanai Sanponpute.

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Uthaiwat, P., Leeudomwong, T. & Sanponpute, T. The comparison of flow rate calibration methods for high-performance liquid chromatography (HPLC) pump. Accred Qual Assur 29, 205–214 (2024). https://doi.org/10.1007/s00769-024-01580-3

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