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Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure-A Case Study in Murine Liver.

ABSTRACT: Lipidomics aim to quantify lipid species in all kinds of samples, including tissues. To subject a fixed amount of sample to various workflows, tissue homogenates were frequently prepared at defined concentrations in water or by addition of organic solvents. Here, we investigated this first step of tissue lipidomics by quantitative flow injection analysis coupled to Fourier-Transform mass spectrometry (FTMS). The influence of sample concentration, solvent composition, and homogenization procedure on the recovery of lipids was studied in murine liver. Liver homogenates were prepared either by grinding tissue in liquid nitrogen or by bead-based homogenization. Ground samples were dissolved at different concentrations in water, methanol, and water/methanol = 1/1 (v/v). Here, lipid recovery depends on solvent composition and sample concentration. The recovery of nonpolar lipid classes, including triglycerides and cholesteryl ester, was decreased in methanolic homogenates. In contrast, due to superior dispersion of precipitates, bead-based homogenization resulted in efficient lipid recovery independent of the solvent composition. However, lipid distribution within samples, i.e., lipid content of supernatant and pellet following centrifugation, was altered substantially by solvent composition. In conclusion, accurate lipid quantification of tissue homogenates requires evaluation of solvent composition, sample concentration, as well as the homogenization method to guarantee efficient lipid recovery. Due to a potential loss of lipids, removal of precipitates by centrifugation prior to lipid extraction should be avoided.

SUBMITTER: Horing M

PROVIDER: S-EPMC8228350 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Feedback from radiologists indicated that differences in image appearance and noise impeded reading of post-contrast computed tomography (CT) scans from an updated CT scanner that was recently added to a fleet of existing scanners from the same vendor, despite using identically named reconstruction algorithms. The goals of this work were to quantify and possibly standardize image quality on the new and an existing scanner using phantom images. Three months of daily quality control images were analyzed to determine the mean CT number and noise magnitude in a water phantom. Next, subtraction images from the uniformity section of an American College of Radiology CT phantom were used to generate noise power spectra for both scanners. Then, a semi-anthropomorphic liver phantom was imaged with both scanners in triplicate using identical body protocols to quantify differences CT number and noise magnitude. Finally, the scanner dependence of CT number and noise magnitude on material attenuation was quantified using a multi-energy CT phantom with 15 material inserts. Significant differences between scanners were determined using a paired or Welch's t test as appropriate. In daily quality control images, the new scanner exhibited slightly higher CT number (0.697 vs. 0.412, P < 0.001, n = 85) and slightly lower noise magnitude (4.85 vs. 4.94, P < 0.001, n = 85). Measured NPS was not significantly different between the existing and new scanners. Interestingly, it was observed that the noise magnitude from the new scanner increased with increasing material attenuation in both the liver (P = 0.008) and multi-energy (P < 0.001) phantoms. Using an alternate reconstruction algorithm with the new scanner eliminated this deviation at high material attenuations. While standard noise evaluation in a water phantom was unable to discern differences between the scanners, more comprehensive testing with higher attenuation materials allowed for the characterization and homogenization of image quality.

Dataset Information

Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure-A Case Study in Murine Liver.

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