LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis
Abstract 20(S)-Protopanaxadiol (PPD) is a saponin derivative of ginsenoside, with more potent biological and pharmacological activities than Rg3 and Rh2. The lack of ionizable centers leads to low mass spectrometry reactions and internal cleavage of three hydroxyl groups, making it challenging to es...
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Nature Portfolio
2025-05-01
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| Online Access: | https://doi.org/10.1038/s41598-025-01432-1 |
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| author | Pengfei Li Shanshan Liu Hongyan Zhuang Mengxi niu Fei Pan Nan Wang Sha Sha Qian Wang Jing Wang |
| author_facet | Pengfei Li Shanshan Liu Hongyan Zhuang Mengxi niu Fei Pan Nan Wang Sha Sha Qian Wang Jing Wang |
| author_sort | Pengfei Li |
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| description | Abstract 20(S)-Protopanaxadiol (PPD) is a saponin derivative of ginsenoside, with more potent biological and pharmacological activities than Rg3 and Rh2. The lack of ionizable centers leads to low mass spectrometry reactions and internal cleavage of three hydroxyl groups, making it challenging to establish highly sensitive PPD mass spectrometry methods. The aim of this study is to establish and validate a quantitative detection method for PPD in multiple matrices using mass spectrometry. The methods used Rh2 as the internal standard and organic solvent liquid–liquid extraction under alkaline conditions for biological sample pretreatment. Isometric separation was achieved through methanol, acetonitrile, and a 10 mmol/L solution of acetic acid (45:45:10, v/v/v) at a flow rate of 0.4 mL/min. Finally, perform mass spectrometry quantification. Comprehensive method validation was conducted on rat plasma samples, and partial method validations were performed on three types of rat tissues (adipose tissue, smooth muscle, and skeletal muscle), bile, urine, fecal samples, and dog plasma samples. The results were in accordance with the requirements of NMPA for bioanalytical method validation, ensuring the accuracy and reliability of our analytical measurements. This study employed a conventional liquid–liquid extraction sample pretreatment scheme, utilizing multiple biological matrices commonly found in a single treatment protocol and liquid chromatography-tandem mass spectrometry detection parameters. The consistency of processing and detection across diverse samples eliminated the need for methodological changes, providing exceptional convenience. Up to 90% of the organic phase and a 50 mm short chromatographic column achieved rapid and effective separation of PPD. A key aspect of our work is the use of a “programmed injection” technique, which significantly reduces the analysis time from 4.2 min during method exploration to 2.4 min. These methods have achieved a relatively low quantification limit of 2.5 ng/mL. The methods established were successfully applied to the kinetic process of PPD in rats, and the pharmacokinetic characteristics of PPD in dogs were studied for the first time. |
| format | Article |
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| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-6acb85dbb96142af91fe7365d0e324bb2025-08-20T03:10:13ZengNature PortfolioScientific Reports2045-23222025-05-011511810.1038/s41598-025-01432-1LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysisPengfei Li0Shanshan Liu1Hongyan Zhuang2Mengxi niu3Fei Pan4Nan Wang5Sha Sha6Qian Wang7Jing Wang8Beijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersBeijing Anding Hospital, Capital Medical University/National Clinical Research Center for Mental Disorders/National Center for Mental Disorders/Beijing Key Laboratory of Mental DisordersAbstract 20(S)-Protopanaxadiol (PPD) is a saponin derivative of ginsenoside, with more potent biological and pharmacological activities than Rg3 and Rh2. The lack of ionizable centers leads to low mass spectrometry reactions and internal cleavage of three hydroxyl groups, making it challenging to establish highly sensitive PPD mass spectrometry methods. The aim of this study is to establish and validate a quantitative detection method for PPD in multiple matrices using mass spectrometry. The methods used Rh2 as the internal standard and organic solvent liquid–liquid extraction under alkaline conditions for biological sample pretreatment. Isometric separation was achieved through methanol, acetonitrile, and a 10 mmol/L solution of acetic acid (45:45:10, v/v/v) at a flow rate of 0.4 mL/min. Finally, perform mass spectrometry quantification. Comprehensive method validation was conducted on rat plasma samples, and partial method validations were performed on three types of rat tissues (adipose tissue, smooth muscle, and skeletal muscle), bile, urine, fecal samples, and dog plasma samples. The results were in accordance with the requirements of NMPA for bioanalytical method validation, ensuring the accuracy and reliability of our analytical measurements. This study employed a conventional liquid–liquid extraction sample pretreatment scheme, utilizing multiple biological matrices commonly found in a single treatment protocol and liquid chromatography-tandem mass spectrometry detection parameters. The consistency of processing and detection across diverse samples eliminated the need for methodological changes, providing exceptional convenience. Up to 90% of the organic phase and a 50 mm short chromatographic column achieved rapid and effective separation of PPD. A key aspect of our work is the use of a “programmed injection” technique, which significantly reduces the analysis time from 4.2 min during method exploration to 2.4 min. These methods have achieved a relatively low quantification limit of 2.5 ng/mL. The methods established were successfully applied to the kinetic process of PPD in rats, and the pharmacokinetic characteristics of PPD in dogs were studied for the first time.https://doi.org/10.1038/s41598-025-01432-120 (S)-protopanaxadiolLC–MS/MSPharmacokineticBiological matricesProgrammed injection |
| spellingShingle | Pengfei Li Shanshan Liu Hongyan Zhuang Mengxi niu Fei Pan Nan Wang Sha Sha Qian Wang Jing Wang LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis Scientific Reports 20 (S)-protopanaxadiol LC–MS/MS Pharmacokinetic Biological matrices Programmed injection |
| title | LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| title_full | LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| title_fullStr | LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| title_full_unstemmed | LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| title_short | LC–MS/MS quantification of 20(S)-protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| title_sort | lc ms ms quantification of 20 s protopanaxadiol in complex biological matrices for bioanalytical method validation and pharmacokinetic analysis |
| topic | 20 (S)-protopanaxadiol LC–MS/MS Pharmacokinetic Biological matrices Programmed injection |
| url | https://doi.org/10.1038/s41598-025-01432-1 |
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