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| Sensitive and efficient determination of endogenous multiple N-acyl homoserine lactones in bacteria by liquid chromatography-mass spectrometry |
| Received:October 21, 2024 |
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| KeyWord:N-acyl homoserine lactones(AHLs);bacteria;group sensing(QS);high performance liquid chromatography-mass spectrometry(HPLC-MS) |
| Author Name | Affiliation | E-mail | | ZHAO Wenjie | Agro-Environmental Protection Institute, Ministry of Agriculture and Rural Affairs, Tianjin 300191, China | | | CHEN Peizhen | Agro-Environmental Protection Institute, Ministry of Agriculture and Rural Affairs, Tianjin 300191, China | chenpeizhen2024@163.com | | ZHANG Yanwei | Agro-Environmental Protection Institute, Ministry of Agriculture and Rural Affairs, Tianjin 300191, China | | | LI Shaopeng | Tianjin Agricultural University, Tianjin, 300384, China | lishaopeng518896@163.com |
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| Abstract: |
| In order to develop a high-performance liquid chromatography-mass spectrometry（HPLC-MS） method for quantitatively detecting endogenous low concentrations of N-acyl homoserine lactones（AHLs）in bacteria, this study extracted AHLs produced by bacteria through liquid-liquid extraction, optimized mass spectrometry and chromatographic conditions, and established a detection system with good linear relationship, small matrix effect, and high recovery rate. The results showed that the linear correlation coefficients（r）of the seven AHLs were all greater than 0.995, and the r values of all AHLs were greater than 0.990, indicating that the method has a good linear relationship. The substrate effects of 8 types of AHLs are all greater than 80.0%, and the bacterial growth matrix has a relatively small impact on the detection of endogenous AHLs. The study also found that AHLs with longer carbon chains have less interference in complex matrices, while AHLs containing carbonyl groups exhibit a certain signal suppression effect. The recovery rates of 7 types of AHLs all exceeded 80.0%, indicating that this method has high recovery rates and good accuracy and reliability. The method was used to detect endogenous AHLs in bacteria, and the results showed that C10-HSL and C12-HSL were the main AHLs, further verifying the effectiveness of the method. This method can accurately detect endogenous trace AHLs in bacteria, providing reliable technical support for studying endogenous AHLs in bacterial quorum sensing（QS）systems. |
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