Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA216213

Local Sample ID:	Free Drug 04_1_14_1_949
Subject ID:	SU002334
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Treatment:

Treatment ID:	TR002346
Treatment Summary:	The extraction of metabolites from FaDu cells was carried out following a similar procedure used in protein extraction. After treatment with PTX (40μM), PTX-PLGA NPs (40 μM), drug-free PLGA NPs and culture media, PBS replaced media and the cells were collected from the dishes using cell scrapers. The detached cell pellets were then collected by centrifugation at 15000 rpm for 10 min at 4oC using Universal 320R Benchtop Centrifuge (Hettich, Beverly, MA, USA) to remove excess PBS. To release a broad range of metabolites, the collected cell pellets were first treated with 0.1% FA in methanol followed by sonication using probe ultrasonicator Q500 (Terra Universal, Inc, Fullerton, CA, USA)for 30 sec at 30% amplitude 16. The samples were then centrifuged at 15,000 rpm for 10 minutes at 4oC, and the upper phase was transferred into LC vials for further drying using a Genevac evaporator (SP Industries, Warminster, PA, USA). The reaction was stopped with 1% formic acid (FA) followed by drying using a Genevac evaporator. Dried samples were stored at −80 °C for further LC-MS/MS analysis.

Treatment ID:

TR002346

Treatment Summary:

The extraction of metabolites from FaDu cells was carried out following a similar procedure used in protein extraction. After treatment with PTX (40μM), PTX-PLGA NPs (40 μM), drug-free PLGA NPs and culture media, PBS replaced media and the cells were collected from the dishes using cell scrapers. The detached cell pellets were then collected by centrifugation at 15000 rpm for 10 min at 4oC using Universal 320R Benchtop Centrifuge (Hettich, Beverly, MA, USA) to remove excess PBS. To release a broad range of metabolites, the collected cell pellets were first treated with 0.1% FA in methanol followed by sonication using probe ultrasonicator Q500 (Terra Universal, Inc, Fullerton, CA, USA)for 30 sec at 30% amplitude 16. The samples were then centrifuged at 15,000 rpm for 10 minutes at 4oC, and the upper phase was transferred into LC vials for further drying using a Genevac evaporator (SP Industries, Warminster, PA, USA). The reaction was stopped with 1% formic acid (FA) followed by drying using a Genevac evaporator. Dried samples were stored at −80 °C for further LC-MS/MS analysis.