Improved Folch Method for Liver-Fat Quantification
Fatty liver represents a significant metabolic pathology of excess intrahepatic fat in domestic animals and humans. Quantification of hepatic-fat content is therefore essential for diagnosis and investigation of liver and metabolic disease. However, the reproducibility of hepatic steatosis analysis...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-01-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2020.594853/full |
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| author | Ramgopal Mopuri Mugagga Kalyesubula Mugagga Kalyesubula Alexander Rosov Nir Edery Uzi Moallem Hay Dvir |
| author_facet | Ramgopal Mopuri Mugagga Kalyesubula Mugagga Kalyesubula Alexander Rosov Nir Edery Uzi Moallem Hay Dvir |
| author_sort | Ramgopal Mopuri |
| collection | DOAJ |
| description | Fatty liver represents a significant metabolic pathology of excess intrahepatic fat in domestic animals and humans. Quantification of hepatic-fat content is therefore essential for diagnosis and investigation of liver and metabolic disease. However, the reproducibility of hepatic steatosis analysis is often low due to subjective and technical factors. We hypothesized that improvement in tissue-lipids extraction efficiency would contribute to the accuracy and precision of liver-fat determination. To test it, we investigated the effect of standardized tissue sonication on liver-fat quantification by the Folch method in sheep. Liver samples from grownup lambs of lean (n = 16) and fatty (n = 15) livers, and from pregnant ewes (n = 6) who died from pregnancy toxemia (PT), were used for hepatic-fat content determination with or without tissue sonication. In the grown lambs, an average hepatic-fat content of 6.6% was determined in sonicated compared to 5.1% in non-sonicated specimens (P = 0.0002). Similarly, in ewes with PT, an average of 12.5% was determined with sonication compared to 10.8% without it (P = 0.0006), and the reproducibility was higher with sonication (CV of 3.1 vs. 6.1%, respectively). Thus, tissue sonication improved the efficiency of liver-lipids extraction and was significant to the accuracy and precision of hepatic-fat determination. Enzymatic quantification of triglycerides was moderately correlated with the results obtained gravimetrically (r = 0.632, P < 0.005). The reported data provide reliable reference values for pregnancy toxemic sheep. The significant improvement in liver-fat quantification observed with the reported revised protocol is likely applicable to most mammals and humans. |
| first_indexed | 2024-12-16T11:14:41Z |
| format | Article |
| id | doaj.art-1a3e50fa9a8e4cf496ff301282716ace |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-12-16T11:14:41Z |
| publishDate | 2021-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj.art-1a3e50fa9a8e4cf496ff301282716ace2022-12-21T22:33:37ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692021-01-01710.3389/fvets.2020.594853594853Improved Folch Method for Liver-Fat QuantificationRamgopal Mopuri0Mugagga Kalyesubula1Mugagga Kalyesubula2Alexander Rosov3Nir Edery4Uzi Moallem5Hay Dvir6Volcani Center—Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, IsraelVolcani Center—Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, IsraelDepartment of Animal Science, The Hebrew University of Jerusalem, Rehovot, IsraelVolcani Center—Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, IsraelPathology Laboratory, Kimron Veterinary Institute, Veterinary Services, Rishon LeZion, IsraelVolcani Center—Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, IsraelVolcani Center—Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, IsraelFatty liver represents a significant metabolic pathology of excess intrahepatic fat in domestic animals and humans. Quantification of hepatic-fat content is therefore essential for diagnosis and investigation of liver and metabolic disease. However, the reproducibility of hepatic steatosis analysis is often low due to subjective and technical factors. We hypothesized that improvement in tissue-lipids extraction efficiency would contribute to the accuracy and precision of liver-fat determination. To test it, we investigated the effect of standardized tissue sonication on liver-fat quantification by the Folch method in sheep. Liver samples from grownup lambs of lean (n = 16) and fatty (n = 15) livers, and from pregnant ewes (n = 6) who died from pregnancy toxemia (PT), were used for hepatic-fat content determination with or without tissue sonication. In the grown lambs, an average hepatic-fat content of 6.6% was determined in sonicated compared to 5.1% in non-sonicated specimens (P = 0.0002). Similarly, in ewes with PT, an average of 12.5% was determined with sonication compared to 10.8% without it (P = 0.0006), and the reproducibility was higher with sonication (CV of 3.1 vs. 6.1%, respectively). Thus, tissue sonication improved the efficiency of liver-lipids extraction and was significant to the accuracy and precision of hepatic-fat determination. Enzymatic quantification of triglycerides was moderately correlated with the results obtained gravimetrically (r = 0.632, P < 0.005). The reported data provide reliable reference values for pregnancy toxemic sheep. The significant improvement in liver-fat quantification observed with the reported revised protocol is likely applicable to most mammals and humans.https://www.frontiersin.org/articles/10.3389/fvets.2020.594853/fullfatty liverpregnancy toxemiatriglycerideshepatic fat quantificationlipids extractionsteatosis |
| spellingShingle | Ramgopal Mopuri Mugagga Kalyesubula Mugagga Kalyesubula Alexander Rosov Nir Edery Uzi Moallem Hay Dvir Improved Folch Method for Liver-Fat Quantification Frontiers in Veterinary Science fatty liver pregnancy toxemia triglycerides hepatic fat quantification lipids extraction steatosis |
| title | Improved Folch Method for Liver-Fat Quantification |
| title_full | Improved Folch Method for Liver-Fat Quantification |
| title_fullStr | Improved Folch Method for Liver-Fat Quantification |
| title_full_unstemmed | Improved Folch Method for Liver-Fat Quantification |
| title_short | Improved Folch Method for Liver-Fat Quantification |
| title_sort | improved folch method for liver fat quantification |
| topic | fatty liver pregnancy toxemia triglycerides hepatic fat quantification lipids extraction steatosis |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2020.594853/full |
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