The Novel Method For Identification Of The Botanic Origin Of Honey And Its Hepatoprotection

Honey is a naturally sweet substance, produced by the Apismellifera bee from the nectar or secretions of plants and stored in honeycomb. In recent years, many researchers in analytical chemistry and food science began to pay attention to identify the botanical origin of the honey and its health effects. The fingerprint of jujube honey, sour jujube honey and lychee honey focused on HPLC-DAD was mainly established in this paper. Combined with chemometric method, a new method for identificating the three honeys was developed. At the same time the antioxidant activities and hepatoprotection of jujube honey and buckwheat honey were also studied in this paper. This study enriched the identification of the botanical origin of the honey, and provided the basis for the development and utilization of honey. The full text was divided into four chapters, and the main contributions are as follows:1. Established anew method for identification of the botanical origin of the jujube honey.(1) The headspace solid-phase microextraction-gas chromatography mass spectrometry (HS-SMPE-GC-MS) was used for determining the aromatic constituent of jujube honey, vitex honey, rape honey and linden honey. The results showed that 92 aromatic compounds were determined in the four honeys. The contents of aromatic aldehydes and ketones in jujube honey and vitex honey were higher than rape honey and linden honey. However, the contents of aromatic acid in rape honey and linden honey were higher than jujube honey and vitex honey. Aromatic compounds in jujube honey from Shaanxi Jianxian were significantly different from others. Its aromatic compounds of aldehyde and ketone were lower than others. This could lay foundation for fingerprint of the aromatic constituents.(2) The fingerprint of jujube honey, sour jujube honey and lychee honey focused on HPLC-DAD was established. And combination with principal component analysis, a new method for indentifying the botanical oringin of the three honeys was established. The results showed that 31,44,42 common peaks were found in the fingerprints of jujube honeys, sour jujube honeys and lychee honeys, respectively. Twenty two common peaks were found in all honey samples, in which 19 common peaks and 7 phenolic compounds were used as variables for classification. All 30 honeys were correctly classified according to their botanical origin using a chemometric method. The overall correct classification rate reached 100%.2. Investigated the mechanism of antioxidant and hepatoprotection of the honeys. (1) The buckwheat honey, jujube honey and the extract of Crataegus pinnatifida pollen were studied for their phenolic compounds, antioxidant activities, and their protection on pBR 322 plasmid DNA and mice lymphocytes. The results showed that eight phenolic compounds were found in buckwheat honey, in which rutin and quercetin were the majority. The total phenolic content of buckwheat honey was 2.039 mg GA/g. Therefore, buckwheat honey owned strong scavenging DPPH radical activity, super reducing power and chelating capability. Jujube honey samples from different origin had different total phenolic content and antioxidant activities. Jujube honey from Shanxi Linxian owned higher total phenolic content and super antioxidant activities. Therefore, it showed good protection on the pBR322 DNA damage, and the percentage of supercoiled DNA was 84.57%. The total phenolic content and total flavone content of the extract of Crataegus pinnatifida pollen were 17.65 mg GA/g and 8.04 mg rutin/g, respectively. Thus it owned super reducing power and chelating capability. It protected pBR322 DNA from oxidative damage through chelating Fe+ of Fenton reaction. And it protected mice lymphocytes against H2O-induced DNA damage through scavenging H2O2 and hydroxyl radicals, chelating Fe2+. The studies of antioxidant in vitro indicated that buckwheat honey, jujube honey and the extract of Crataegus pinnatifida pollen possessed phenolic compound, strong antioxidant activities and good protection on DNA damage incuced by oxidative stress.(2) The mechanism of hepatoprotection of buckwheat honey and jujube honey was studied. Pretreatment with buckwheat honey at the dose of 0.22 g/10 g BW, twice daily, for 10 weeks, the serum lipoprotein antioxidation and oxygen radical absorbance capacity (ORAC) antioxidant capability were increased significantly (P<0.05). After intraperitoneal injection with CCl4, the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were decreased (P<0.05), the production of malondialdehyde was inhibited and the activities of antioxidant enzymes was increased by administration with buckwheat honey. At the same time, we found pretreatment with buckwheat honey can inhibite lymphocytes DNA damage induced by CCl4. The histological observations showed administration with buckwheat honey ameliorate liver swelling, lessen sinusoidal spaces, reduce inflammatory cell infiltration and decrease hepatocyte ballooning degeneration. Administration with jujube honey at the dose of 54.0 g/kg/d, for 12 weeks, the antioxidant capability of serum lipoprotein was increased (P<0.05), the serum ALT and AST activities were decreased (P<0.05), the production of malondialdehyde was inhibited and the activities of GSH-Px was increased significantly (P<0.05). Admindistration with jujube honey at the dose of 54.0 g/kg/d completely inhibited the generation of 8-hydroxy-2-deoxyguanosine (8-OHdG). The histological observations supported the hepatoprotection of jujube honey. All the animal studies indicated that buckwheat honey showed super protection on CCl4 induced acute liver injury. The hepatoprotective effect may be related to its increasing antioxidant activity. Jujube honey can protect liver from chronic alcohol-induced liver damge in mice. The hepatoprotection should be related to its increasing serum antioxidant activity and inhibing lipid peroxidation of hepatocyte induced by chronic alcohol.