PMID- 37534560 OWN - NLM STAT- MEDLINE DCOM- 20230804 LR - 20230809 IS - 1872-2059 (Electronic) IS - 1000-8713 (Print) IS - 1000-8713 (Linking) VI - 41 IP - 8 DP - 2023 Aug TI - [Fingerprint of sophorolipids based on ultra-high performance liquid chromatography-charged aerosol detection]. PG - 722-729 LID - 10.3724/SP.J.1123.2022.12025 [doi] AB - Sophorolipids are secondary metabolites produced during fermentation by nonpathogenic yeasts. These molecules are amphiphilic and consist of a hydrophilic sophora sugar moiety and a hydrophobic hydroxylated fatty acid. Based on their degree of esterification, sophorolipids can be divided into the acid and lactone types. Sophorolipids are highly promising biosurfactants with good antibacterial, antiviral, and other biological activities. Moreover, they are characterized by mildness, low toxicity, and environmental friendliness. However, their composition is quite complex, and effective methods for their quality evaluation are lacking. Since sophorolipids do not absorb ultraviolet (UV) light, common UV detectors are unsuitable for fingerprint establishment. In this study, we first selected a charged aerosol detector (CAD) to establish the ultra-high performance liquid chromatography (UHPLC) fingerprint of sophorolipids. The detector had high sensitivity, good reproducibility, and excellent suitability for the detection of substances with no or weak ultraviolet absorption. We then evaluated the similarities between 17 batches of sophorolipid samples. The samples were extracted by ultrasound for 10 min in 80% ethanol aqueous solution at a liquid-solid ratio of 10ratio1 (mL/g) and then separated on a Thermo Fisher Scientific Hypersil Gold chromatographic column (150 mmx2.1 mm, 1.9 mum). Separation was performed using acetonitrile-0.01% (v/v) formic acid aqueous solution as the mobile phase via gradient elution. The flow rate was 0.2 mL/min, and the column temperature was 40 ℃. The CAD was used under the following conditions: power function of 1.0, data rate of 5 Hz, filter constant of 3.6, and evaporation temperature of 45 ℃. The chromatograms and retention times of the sophorolipids were compared, and 16 common peaks with strong responses, good resolutions, and stable retention times were selected as characteristic peaks. Oleic acid was chosen as the reference peak because it achieved good separation and a strong chromatographic response in all batches of samples. UHPLC-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was used to identify chromatographic peaks in the sophorolipid fingerprints. The results were combined with the retention time rule of the sophorolipids, leading to their identification based on matching with the results of the primary database, the precise relative molecular mass and fragmentation rule of secondary fragments, a self-built database, and the PubChem database. Sixteen compounds were identified, including eight acid sophorolipids, six lactone sophorolipids, and two aliphatic acids. The results of precision, repeatability, and 24 h stability tests indicated that the relative standard deviations (RSDs) of the retention times and peak areas of the 15 characteristic peaks relative to the control peak (oleic acid) were less than 3.0% (n=6). Seventeen batches of sophorolipid samples were analyzed, and the similarity values of all fingerprints were found to be 0.965 or higher. Little differences in chemical composition were observed among the different batches of sophorolipid samples, and the quality of the sophorolipids was relatively consistent. The fingerprint established in this study is stable and reliable; it can be used for the quality evaluation of sophorolipids and lays a solid foundation for future research on production technology and the development and utilization of sophorolipids. The successful application of a universal CAD to the fingerprint establishment of sophorolipids also provides a reliable solution for the fingerprint establishment of substances with no or weak ultraviolet absorption. FAU - Cao, Qin-Ling AU - Cao QL AD - Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China. FAU - Zhao, Xiao-Dan AU - Zhao XD AD - Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China. FAU - Shen, Guo-Bin AU - Shen GB AD - Thermo Fisher Scientific (China) Co., Ltd., Shanghai 201206, China. FAU - Wang, Zhu-Qin AU - Wang ZQ AD - Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China. FAU - Zhang, Hong-Yang AU - Zhang HY AD - Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China. FAU - Zhang, Min AU - Zhang M AD - Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, School of Pharmaceutical Sciences, East China University of Science and Technology, Shanghai 200237, China. FAU - Hu, Ping AU - Hu P AD - Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China. LA - chi PT - English Abstract PT - Journal Article PL - China TA - Se Pu JT - Se pu = Chinese journal of chromatography JID - 9424804 RN - 0 (sophorolipid) RN - 0 (Oleic Acids) SB - IM MH - Chromatography, High Pressure Liquid MH - Reproducibility of Results MH - Quality Control MH - *Oleic Acids PMC - PMC10398824 OTO - NOTNLM OT - charged aerosol detector (CAD) OT - fingerprint OT - ultra-high performance liquid chromatography (UHPLC) EDAT- 2023/08/03 06:43 MHDA- 2023/08/04 06:43 PMCR- 2023/08/08 CRDT- 2023/08/03 05:39 PHST- 2023/08/04 06:43 [medline] PHST- 2023/08/03 06:43 [pubmed] PHST- 2023/08/03 05:39 [entrez] PHST- 2023/08/08 00:00 [pmc-release] AID - 10.3724/SP.J.1123.2022.12025 [doi] PST - ppublish SO - Se Pu. 2023 Aug;41(8):722-729. doi: 10.3724/SP.J.1123.2022.12025.