Comparison of simple monophasic versus classical biphasic extraction protocols for comprehensive UHPLC-MS/MS lipidomic analysis of Hela cells

Calderón, Carlos; Sanwald, Corinna; Schlotterbeck, Jörg; Drotleff, Bernhard; Lämmerhofer, Michael

doi:10.1016/j.aca.2018.10.035

Cited by 39 publications

(26 citation statements)

References 33 publications

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“…Analyses were performed by Agilent 1290 Series UHPLC instrument (Agilent, Waldbronn, Germany) coupled to Sciex TripleTOF 5600+ MS (Sciex, Concord, Ontario; Canada) with duospray source and Pal HTC‐XS autosampler from CTC (Zwingen, Switzerland). Positive and negative ESI ionization were used in separate LC–MS runs with the same chromatographic separation conditions as described previously . Briefly: Acquity UPLC CSH C18 (130 Å, 1.7 µm, 2.1 mm × 100 mm) column was utilized with Acquity UPLC CSH C18 VanGuard pre‐column (130 Å, 1.7 µm, 2.1 mm × 5 mm) (Waters, Eschborn, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Lipid extraction was performed with 1 million cells using IPA:H 2 O (90:10 v/v) as described previously [30] (see details of extraction protocol in Supporting Information A1).…”

Section: Significance Statementmentioning

confidence: 99%

“…Positive and negative ESI ionization were used in separate LC-MS runs with the same chromatographic separation conditions as described previously. [27,30] Briefly: Acquity UPLC CSH C18 (130 Å, 1.7 µm, 2.1 mm × 100 mm) column was utilized with Acquity UPLC CSH C18 VanGuard pre-column (130 Å, 1.7 µm, 2.1 mm × 5 mm) (Waters, Eschborn, Germany). The mobile phase was composed of 10 mm ammonium formate and 0.1% formic acid dissolved in 60:40 ACN: MS/MS data were obtained by DIA using SWATH.…”

Section: Lc-ms Measurementmentioning

confidence: 99%

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Lipid Atlas of Keratinocytes and Betulin Effects on its Lipidome Profiled by Comprehensive UHPLC–MS/MS with Data Independent Acquisition Using Targeted Data Processing

et al. 2019

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Betulin is a pentacyclic triterpene with demonstrated healing properties in mid-dermal wounds. A few earlier studies have provided insights into the wound healing effects on the molecular level. However, there are still questions left on the molecular targets of betulin. Therefore, a pharmacolipidomics analysis of betulin is undertaken in human immortalized keratinocytes to monitor alterations in the lipid profiles induced by treatment with betulin. For this purpose, lipid extracts of keratinocytes treated with betulin and untreated controls are comprehensively analyzed by an untargeted UHPLC-ESI-QTOF-MS/MS lipidomics profiling workflow using data-independent acquisition. Targeted data processing allows the identification of 611 lipid species from 21 different lipid classes. Statistical analysis of the identified lipids shows significant changes in 440 lipid species that can be described as downregulation of cholesteryl esters and triacylglycerides and upregulation of glycerophospholipids, sphingolipids, and diacylglycerides. Additionally, some other signals corresponding to triterpenes are found in the betulin group and suggested that betulin is incorporated (in the membrane) and metabolized in keratinocytes.

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Section: Methodsmentioning

confidence: 99%

“…Lipid extraction was performed with 1 million cells using IPA:H 2 O (90:10 v/v) as described previously [30] (see details of extraction protocol in Supporting Information A1).…”

Section: Significance Statementmentioning

confidence: 99%

Section: Lc-ms Measurementmentioning

confidence: 99%

See 1 more Smart Citation

Lipid Atlas of Keratinocytes and Betulin Effects on its Lipidome Profiled by Comprehensive UHPLC–MS/MS with Data Independent Acquisition Using Targeted Data Processing

et al. 2019

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“…Lipids were extracted by using an IPA-based protein precipitation approach (Calderón et al, 2019;Sarafian et al, 2014) with minor modifications. Lysophosphatidylcholine (lysoPC [19:0]), phosphati dyl choline (PC [19:0/19:0]), phosphatidylethanolamine (PE [17:0/17:0]), ceramide (Cer [d18:1/17:0]), and triacylglycerol (TG [15:0/18:1(d7)/15:0]) were added to each sample before extraction to serve as internal standards.…”

Section: Extraction Of Lipidsmentioning

confidence: 99%

Remodeling of Arctic char (Salvelinus alpinus) lipidome under a stimulated scenario of Arctic warming

Wang

Gong

Deng

et al. 2021

Global Change Biology

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Arctic warming associated with global climate change poses a significant threat to populations of wildlife in the Arctic. Since lipids play a vital role in adaptation of organisms to variations in temperature, high‐resolution mass‐spectrometry‐based lipidomics can provide insights into adaptive responses of organisms to a warmer environment in the Arctic and help to illustrate potential novel roles of lipids in the process of thermal adaption. In this study, we studied an ecologically and economically important species—Arctic char (Salvelinus alpinus)—with a detailed multi‐tissue analysis of the lipidome in response to chronic shifts in temperature using a validated lipidomics workflow. In addition, dynamic alterations in the hepatic lipidome during the time course of shifts in temperature were also characterized. Our results showed that early life stages of Arctic char were more susceptible to variations in temperature. One‐year‐old Arctic char responded to chronic increases in temperature with coordinated regulation of lipids, including headgroup‐specific remodeling of acyl chains in glycerophospholipids (GP) and extensive alterations in composition of lipids in membranes, such as less lyso‐GPs, and more ether‐GPs and sphingomyelin. Glycerolipids (e.g., triacylglycerol, TG) also participated in adaptive responses of the lipidome of Arctic char. Eight‐week‐old Arctic char exhibited rapid adaptive alterations of the hepatic lipidome to stepwise decreases in temperature while showing blunted responses to gradual increases in temperature, implying an inability to adapt rapidly to warmer environments. Three common phosphatidylethanolamines (PEs) (PE 36:6|PE 16:1_20:5, PE 38:7|PE 16:1_22:6, and PE 40:7|PE 18:1_22:6) were finally identified as candidate lipid biomarkers for temperature shifts via machine learning approach. Overall, this work provides additional information to a better understanding of underlying regulatory mechanisms of the lipidome of Arctic organisms in the face of near‐future warming.

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“…In order to ensure that they have stable clinical therapeutic effect, reasonable quality control is very necessary. HPLC fingerprint chromatography provides a more comprehensive approach for quality control of HWDs because of its wide applicability, high sensitivity, high efficiency, and easy operation [19,20]. Similarity analysis, as a significant evaluation index of the fingerprint of TCMs, can reflect the similarity of the response ratio of chemical components and find common components (CCs) among different batches of samples.…”

Section: Introductionmentioning

confidence: 99%

Study on the Material Basis of Houpo Wenzhong Decoction by HPLC Fingerprint, UHPLC-ESI-LTQ-Orbitrap-MS, and Network Pharmacology

et al. 2019

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Houpo Wenzhong Decoction (HWD) as a classical prescription has been widely used for hundreds of years. However, the quality of HWDs is difficult to control because of its herb materials from different regions. It is a new idea to use HPLC fingerprints, LTQ-ESI-Orbitrap-MS, and network pharmacology in combination to screen common components (CCs) as potential quality control indicators. In this paper, the fingerprints of HWDs were established, which were used to determine HWDs compounded from different sources of traditional Chinese medicines (TCMs). Through the similarity analysis, 45 CCs were selected. UHPLC-LTQ-ESI-Orbitrap-MS was used to carry out the chemical composition analysis of HWD. Seventy-three chemical constituents were distinguished, and 30 CCs were identified. Through network pharmacology, networks of candidate CCs, diseases, and candidate targets were constructed. Finally, four CCs were screened as potential active ingredient markers of HWD, and a method for content determination of these four components was established. A rapid, reasonable, and effective method for quality evaluation and control of HWDs was established. It provides a reference for the further development and research of HWDs and a new way of thinking for the research of other Chinese medicine prescriptions.

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Comparison of simple monophasic versus classical biphasic extraction protocols for comprehensive UHPLC-MS/MS lipidomic analysis of Hela cells

Cited by 39 publications

References 33 publications

Lipid Atlas of Keratinocytes and Betulin Effects on its Lipidome Profiled by Comprehensive UHPLC–MS/MS with Data Independent Acquisition Using Targeted Data Processing

Lipid Atlas of Keratinocytes and Betulin Effects on its Lipidome Profiled by Comprehensive UHPLC–MS/MS with Data Independent Acquisition Using Targeted Data Processing

Remodeling of Arctic char (Salvelinus alpinus) lipidome under a stimulated scenario of Arctic warming

Study on the Material Basis of Houpo Wenzhong Decoction by HPLC Fingerprint, UHPLC-ESI-LTQ-Orbitrap-MS, and Network Pharmacology

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