Noninvasive markers in monitoring hepatocellular damage: role of natural antioxidants: nigella sativa, honey and silymarin in treatment of hepatocellular damage

Liver fibrosis induced experimentally by using carbontetrachloride [CC14] as a toxic substance. To evaluate the treatment effects of some natural antioxidants such as Nigella sativa [N. sativa], Honey [H.] and Silymarin [S.] in CC14-induced liver fibrosis in rats, some biochemical markers such as glutathione-S-transferase [alpha-GST], Malonedialdehyde [MDA], Nitric oxide [NO] and matrix metalloproteinase [MMP-2] were determined and compared to the liver functions [ALT and AST] and Hemoglobin [Hb]. 100 rats were divided into 20 normal rats served as control, 20 rats injected 1 mg/kg/day CC14 intrapretonially 3 times a week for 8 weeks, 60 rats divided equally into 3 subgroups subjected to the same injection plus oral administration of 50 mg/kg/ day N. sativa, H. and S. daily for another 4 weeks. alpha-GST, MDA, NO and MMP-2 were estimated by enzyme immunoassay and ALT, AST and Hb were estimated by colorimetric assay. Statistical program SPSS, VI2 was used for data analysis. The results showed a significant increase [P<0.05] in alpha-GST, MDA and NO and high significant increase [P<0.01] in MMP-2 in rats group administered CC14 as compared to normal control. The results matched the role of S. in the treatment of hepatic fibrosis and revealed the role of N.sativa as treatment agent by improvement in ALT and AST, enhance antioxidant capacity by significant decrease [P<0.05] in alpha-GST and NO, improvement in general health by high significant increase [P<0.01] in Hb, enhanced antifibrotic response by high significant decrease [P<0.01] in MMP-2 and decrease in lipid peroxidation by significant decrease [P<0.05] in MDA compared to CC14 group. Although H. showed the least effective ability for treatment of liver fibrosis, it appeared to improve liver functions. The histological results were found to be matched with the biochemical results. This work proved that one of the mechanisms involved in the process of liver fibrogenesis induced by CC14 is the development of oxidative stress and fibrotic response, the determined biochemical markers proved to be good markers for the detection of hepatocellular damage and documented also that N. sativa and H. can be used effectively as tools in the treatment of hepatic injury without any fear of complications

Monitoring of aflatoxins in Egyptian hot chili prepared for export using GPC as a new cleanup technique and reversed phase-HPLC

New method was developed for determination of aflatoxins BI, B2,G1 and G2 in hot chili using gel permeation chromatography [GPC as a new clean up technique. High performance liquid chromatography [HPLC] connected with fluorescence detector was used for determination of the four types of aflatoxin. The method was tested on different levels of fortified samples of hot chili [6 sample for each level] and the limit of detection [LOD] was [0.5 ph]. The average recoveries of aflatoxins from different levels [2,4,10 and 50 ppb] varied between 87 and 107%. The reproducibility expressed as relative standard deviation was less than 20%. The method showed to be linear from the limit of quantitation [2 ug/kg] up to 50 ug/kg level. The four types of aflatoxins were determined in fortified samples by extraction with acetonitrile and purified by GPC, [[Bio bead - SX 3 gerl] was used as stationary phase and hexane: ethyl acetate [1:1] as mobile phase]. Drivatiztion of B1 and G1 were obtained by reaction with trifluoroacetic acid [TEA] in a pre-column step. Quantitation of aflatoxins is carried out using reversed phase high performance liquid chromatography with isocratic mobile phase water/methanol/acetonitrile [65:25:15%]. The detection was done at excitation and emission wavelengths of 360 and 440 nm respectively. This method wss used for monitoring of aflatoxins in 87 hot chili samples prepared for export. Only aflatoxins B1 and B2 were detected and all samples were free from any traces of G1 and G2. Sixty samples were contaminated with aflatoxin B1 and only 20 samples were contaminated with aflatoxin B2. The minimum amount of B1 and B2 was 0.5 ppb LOD], while the maximum amounts were 78.95 and 5.21 ppb respectively. The means were 18.57 and 1.80 ppb for B1 and B2 respectively. Forty six samples were violated with aflatoxin B1 depending on EU MRL limits [2 ppb] when 27 samples were violated depending on CODEX-MIRL limits [5 ppb]