Determination of Paeoniflorin in Lupu Lingling Granules by High Performance Liquid Chromatography - Science Definition

Lupusling granules are a traditional Chinese medicine formulation composed of five key herbal ingredients, including red peony root, peony bark, rehmannia root, and honeysuckle vine. These granules are known for their ability to cool the blood, resolve phlegm, and clear heat while detoxifying the body. The main active component in these granules is paeoniflorin, which has been shown to exert a dual-directional immunomodulatory effect. This makes it an effective treatment option for patients suffering from systemic lupus erythematosus (SLE). To ensure the quality and consistency of Lupusling granules, a high-performance liquid chromatography (HPLC) method was developed to determine the content of paeoniflorin. This analytical technique provides accurate and reliable results, making it suitable for quality control during the production process. The study involved several steps, starting with the preparation of reference and test solutions. A standard solution of paeoniflorin was prepared by accurately weighing 3.04 mg of the reference substance and dissolving it in methanol to make a 10 mL solution. For the test solution, approximately 0.5 g of the granule sample was dissolved in 30 mL of methanol, followed by sonication and filtration. A negative control solution was also prepared, omitting red peony to evaluate potential interferences. Chromatographic conditions were optimized using a Hypersil ODS-2 column with a mobile phase consisting of acetonitrile and 0.4% phosphoric acid (10:90). The detection wavelength was set at 230 nm, with a flow rate of 1.0 mL/min and a column temperature of 25°C. The method was validated through various tests, including linearity, specificity, precision, stability, repeatability, and recovery. The linear relationship between the peak area and the concentration of paeoniflorin was found to be strong, with a correlation coefficient (r) of 0.9999. No interfering peaks were observed in the chromatograms, confirming the method's specificity. Precision testing showed a relative standard deviation (RSD) of 1.19%, indicating excellent reproducibility. Stability testing revealed that the paeoniflorin content remained stable for up to 24 hours, with an RSD of 1.30%. Repeatability tests yielded an average content of 9.73 mg/g of paeoniflorin, with an RSD of 1.24%. In the recovery test, the average recovery rate of paeoniflorin was 97.77%, with a low RSD of 0.62%, demonstrating the accuracy of the method. Finally, the method was applied to analyze different batches of Lupusling granules, and the results indicated that the paeoniflorin content should not exceed 9.7 mg per gram of granules. This HPLC method offers a simple, fast, and reliable way to assess the quality of Lupusling granules. It effectively controls the consistency of the product and ensures that it meets the required standards for clinical use.

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