Ethyl caffeate (EC), octyl caffeate(OC), benzyl caffeate(BC) and phenethyl caffeate(PC) were synthesized and evaluated for scavenging of superoxide anion, nitric oxide radical and 1,1-diphenyl-1-picrylhydrazyl radical (DPPH). Antioxidant activity was investigated with reducing power method. Pooled human liver microsome was used for investigating the effects on cytochrome P450 1A2 (CYP1A2) catalytic activities by using phenacetin as a substrate. Dixon and Cornish-Bowden plots were used for enzyme kinetic analysis. The EC, OC, BC and PC potentially inhibited superoxide anion, nitric oxide and DPPH radicals. IC(50) values of superoxide anion scavenging of EC, OC, BC and PC were 16.42, 79.83, 123.69 and 123.69 µg/ml, respectively. EC was more potent than OC and BC in terms of nitric oxide radical scavenger: IC(50) values of EC, OC and BC were 24.16, 37.34 and 52.64 µg/ml, respectively. In addition, the IC(50) values of EC, OC, BC and PC on DPPH radical scavenging were 70.00, 184.56, 285.34 and 866.54 µg/ ml, respectively. The IC(50) values of EC, OC, BC and PC on phenacetin O-deethylation were 124.98, 111.86, 156.68 and 31.05 µg/ml, respectively. Enzyme kinetics showed that the type of inhibition mechanism was mixed-type. The result of this study shows that caffeic acid ester analogues potentially scavenge free radicals and inhibit catalytic activity of CYP1A2. This may lead to important implications in the prevention of CYP1A2-mediated chemical carcinogenesis.

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From our results, caffeic acid and its amide analogues are in vitro inhibitors of human CYP1A2 catalytic activity and free radical formation. They may be useful to be developed as potential chemopreventive agents that block CYP1A2-mediated chemical carcinogenesis.

BackgroundMethomyl is the most common cause of suicidal death but heroin is the most common cause of accidental death. The problem is to determine the exact cause and manner of death between methomyl or heroin toxicity. The evidence from autopsy includes crime scene investigation, toxicological analysis by liquid chromatography with mass spectrometry, and knowledge of methomyl and heroin intoxication.Case presentationA 35-year-old Thai man and a 30-year-old Thai woman were found showing evidence of cyanosis, with a fine froth around the nose and mouth. Postmortem interval time was 24 hours. According to the police’s and hotel owner’s records, the couple stayed together for 1 day before being found dead in bed, naked, with a foul and a fine froth around the nose and mouth. A methomyl insecticide sachet and a plastic box containing white powder form of heroin were found at the scene. Laboratory tests of the male corpse identified the presence of methomyl in the blood of the stomach and morphine, codeine, methadone, and tramadol in the systemic blood. Blood cholinesterase enzyme activity and morphine concentration was 3416 U/L or 53% (normal 6400 U/L) and 0.058 μg/ml respectively. Laboratory test of the female corpse identified the presence of methomyl in the stomach and blood, and cholinesterase enzyme activity was 1965 U/L or 30.7%.ConclusionsCause of death of the male corpse was deemed to be due to heroin intoxication as the blood concentration of morphine was more than the lethal concentration with a morphine/codeine ratio of more than 1:1. Methomyl intoxication of the male corpse was unlikely to be the cause of death because methomyl systemic blood concentration was found to be very low, < 2.5 μg/ml, and cholinesterase enzyme levels did not indicate lethal activity (< 10–15% of normal). The main problem regarding an insurance claim is that the policy will not pay out in the case of heroin-associated deaths, as it is an addictive drug. The policy would pay out on death by suicide with methomyl insecticide, which was not prohibited by the insurance company after 1 year of insurance. So, it is not clear whether or not the family will receive money from the insurance company.

The purpose of this research was to find out the substance which deactivate α-Amanitin ToxicityThe materials and methods used in the study include analysis with high performance liquid chromatography (HPLC) to:1.Demonstrate the standard α-amanitin at concentrations of 25, 50 and 100 µg/ml 2.Determine the deactivation of α-amanitin with 1) 18% acetic acid 2), calcium hydroxide 40 mg/ml, 3) potassium permanganate 20 mg/ml, 4) sodium bicarbonate 20 mg/ml 3.Report the statistical analysis as the mean ± standard deviation (SD) and paired t-test.The result revealed that potassium permanganate could eliminate 100 percent of the α-amanitin at 25, 50 and 100 µg/ml. Calcium hydroxide, sodium bicarbonate and acetic acid had lower elimination rates at those concentrations: 68.43 ± 2.58 (-71.4, -67.2, -66.7%), 21.48 ± 10.23 (-29.4, -25.2, -9.9%) and 3.21 ± 0.02% (-3.2, -3.2, +1.1%), respectively. The conclusion of this study was suggested that potassium permanganate could be applied as an absorbent substance during gastric lavage in patients with mushroom poisoning. It also might be effective as a cleansing wash for uncooked mushrooms. Investigation of potassium permanganate's ability to absorb α-amanitin in animal models and humans should be considered. .Patients with α-amanitin poisoning can develop severe toxic hepatitis, centrilobular necrosis, liver steatosis, and acute tubulointerstitial necrosis leading to hepatorenal syndrome all of which have a high mortality rate. The most effective treatment is emptying the stomach promptly by performing gastric lavage with 1:2,000 tannic acid or 1:10,000 potassium permanganase plus triggering emesis. A warm saline solution of potassium permaganase can be used as an emergency treatment to reduce toxins in the intestines and stomach. Thus, KMnO 4 , NaHCO 3 , Ca(OH) 2 and acetic acid should be considered as a cleanser for uncooked mushrooms to deactivate α-amanitin toxicity and potentially reduce the mortality rate from liver failure, acute renal failure, respiratory failure and gastro-intestinal haemorrhage [9]. Materials and MethodsIn this study, experiments were performed on α-amanitin at concentrations of 25, 50 and 100 µg/ml eached mixed with 1) 18% acetic acid 2), calcium hydroxide 40 mg/ml, 3) potassium permanganate 20 mg/ml, 4) sodium bicarbonate 20 mg/ml (results analyzed with high performance liquid chromatography (HPLC) with Luna C18 (150 × 4.6 mm I.D., 5 micron) from Phenomenex ® , USA), or 5) mobile phase (a mixture of 0.02 M aqueous ammonium acetate and acetonitrile (88/12, v/v) pH 5.0 at an absorbance of 280 nm). Glacial acetic acid was used to Citation: Narongchai P, Narongchai S (2017) Deactivation Study of α-Amanitin Toxicity in Poisonous Amanita spp. Mushrooms by the Common Substances In Vitro. J Forensic Res 8: 396.