over the last decades toxins and secretions from poisonous and venomous animals have been used as drugs and drug leads for treatment of numerous untreatable human ailments .
leech salivary secretion exert antimicrobial agents and has been reported to be used in treatments of back pain , snake venom serve as anticancer , anti - diabetics and anti - hypertensive agents , secretion from cone snail conus magus used in treatment of chronic pain , while hemolymph from african land snail has been reported for their hepatocurative effect against ccl4 intoxicated rats .
honey bees are members of genus apis , and perennial insect species that can utilize nearly all habitats of the world .
there are about seven species of the honey bees with a total of 44 subspecies .
honeybees produce high - quality foods in the form of honey , building materials in the form of propolis , and chemical defenses inform of bee venom and propolis .
propolis is a sticky honeybee resinous product produced by the honeybees to shut the cracks , and act as moisture and thermo stabilizer in the hive .
bee propolis has been documented for it bacteriostatic , immunomodulatory , anti - inflammatory , anti - tumoral , anti - oxidative , hypotensive , hepatoprotective and pancreato protective , and antibacterial properties as well as for the treatments of atherosclerosis among many other uses .
recently , we also reported the hepatocurative , hematopoietic , and its toxicological effects on serum , and tissues of rats following chronic exposure .
it is worth noted that despite the high activity , propolis is very stable compounds , retaining their potency up to several years .
these stable organic compounds would confer different chemical properties and could be implicated in the biological and toxicological effects of the propolis .
however , the chemical composition of bee propolis has been reported to be varied qualitatively and quantitatively , depending on the environmental plant ecology . since
, propolis is a bee product of plant origin , thus at different geographic locations the source plants might vary with respect to the local flora .
literatures have documented the chemical composition of bee propolis from a different region of the word [ 13 - 20 ] , however , none of this study was on propolis of nigerian origin . with the aimed of bridging the gap in knowledge , and the present study sort to evaluate nigerian bee propolis for its chemical composition .
the identity of the propolis was authenticated by an entomologist in the biological sciences department , federal university of technology , minna , nigeria .
the propolis material was chopped into small pieces and air dried in the shade at room temperature for 2 weeks .
200 g of propolis pellets were percolated in 1600 ml of absolute methanol and subsequently allowed to stand in the shade for 48 h before filtration , using filter paper ( whatman no .
1 ) . the extract concentrate was stored in air - tight vials in the refrigerator at 4c , until needed for bioassay .
methanol extract from nigeria bee propolis was screened preliminary for its phytochemical contents including flavonoids , saponins , alkaloids , tannins , cardiac glycosides , anthraquinones phlobatannins , and steroids according to the methods of sofowora   as described by lawal .
the gc / ms analysis of methanol extract from nigerian bee propolis was perform using gc - ms clarus 500 per kin elmer system comprising an aoc-20i auto sampler .  the instrument is equipped with a vf 5 ms fused silica capillary column of 30 m length , 0.25 mm diameter and 0.25 m film thickness . 
the temperatures employed were ; column oven temperature 80c , injection temp 250c at a pressure of 108.0 kpa , with total flow and column flow of 6.20 ml / min and 1.58 ml / min , respectively .
the linear velocity was 46.3 cm / s and a purge flow of 3.0 ml / min .
the gc program ion source and interface temperature were 200.00c and 250.00c , respectively , with solvent cut time of 2.50 min .
the ms program starting time was 3.00 min which ended at 30.00 min with event time of 0.50 s , scan speed of 1666 l / s , scan range 40 - 800 u , and an injection volume of 1 l of the propolis extract ( split ratio 10:1 ) .
the relative percentage of the extract was expressed as percentage with peak area normalization as previously reported by lawal et al . .
interpretation on the mass spectrum was conducted using the database of national institute standard and technology ( nist ) having more than 62,000 patterns .
the fragmentation pattern spectra of the unknown components were compared with those of known components stored in the nist library .
the relative percentage amount of each bio - component was calculated by comparing its average peak area to the total area .
the name , molecular weight , and structure of the components of the test materials were ascertained .
the identity of the propolis was authenticated by an entomologist in the biological sciences department , federal university of technology , minna , nigeria .
the propolis material was chopped into small pieces and air dried in the shade at room temperature for 2 weeks .
200 g of propolis pellets were percolated in 1600 ml of absolute methanol and subsequently allowed to stand in the shade for 48 h before filtration , using filter paper ( whatman no .
the extract concentrate was stored in air - tight vials in the refrigerator at 4c , until needed for bioassay .
methanol extract from nigeria bee propolis was screened preliminary for its phytochemical contents including flavonoids , saponins , alkaloids , tannins , cardiac glycosides , anthraquinones phlobatannins , and steroids according to the methods of sofowora   as described by lawal .
the gc / ms analysis of methanol extract from nigerian bee propolis was perform using gc - ms clarus 500 per kin elmer system comprising an aoc-20i auto sampler .  the instrument is equipped with a vf 5 ms fused silica capillary column of 30 m length , 0.25 mm diameter and 0.25 m film thickness . 
the temperatures employed were ; column oven temperature 80c , injection temp 250c at a pressure of 108.0 kpa , with total flow and column flow of 6.20 ml / min and 1.58 ml / min , respectively .
the linear velocity was 46.3 cm / s and a purge flow of 3.0 ml / min .
the gc program ion source and interface temperature were 200.00c and 250.00c , respectively , with solvent cut time of 2.50 min .
the ms program starting time was 3.00 min which ended at 30.00 min with event time of 0.50 s , scan speed of 1666 l / s , scan range 40 - 800 u , and an injection volume of 1 l of the propolis extract ( split ratio 10:1 ) .
the relative percentage of the extract was expressed as percentage with peak area normalization as previously reported by lawal et al . .
interpretation on the mass spectrum was conducted using the database of national institute standard and technology ( nist ) having more than 62,000 patterns .
the fragmentation pattern spectra of the unknown components were compared with those of known components stored in the nist library .
the relative percentage amount of each bio - component was calculated by comparing its average peak area to the total area .
the name , molecular weight , and structure of the components of the test materials were ascertained .
the result revealed the presence of flavonoids , saponins , alkaloids , tannins , cardiac glycosides , anthraquinones phlobatannins , and steroids in methanol extract of nigeria bee propolis . the gc / ms chromatogram revealed nineteen peaks [ figure 1 ] representing 60 different chemical compounds .
the chemical compounds with their molecular formula , molecular weight , retention time ( rt ) , and % peak area are presented in table 2 while the chemical structures as revealed by the gc / ms were shown in figure 2 .
phytochemical compositions of methanol extract of nigerian bee propolis gas chromatography mass spectrometry chromatogram of methanol extract of nigerian bee ( apis mellifera ) propolis bio - active components identified in methanol extract of nigerian bee ( apis mellifera ) propolis using gcms structures of some chemical compounds identified from methanol extract of nigerian bee propolis by gas chromatography mass spectrometry
the use of and search for , plant - derived drugs have accelerated in recent years .
biochemist , pharmacologists , botanists , microbiologists , and natural - products chemists globally are continuously in search of natural - products for bioactive phytoconstituents that could serve as a drug lead for treatment of various human ailments .
the present study revealed the present of various important phytochemicals in methanol extract from nigerian bee propolis [ table 1 ] .
flavonoids are phenolic compounds with important roles in scavenging free radicals and thus play vital roles in preventing oxidative stress associated disorder .
tannin is non - toxic compounds that are known for there antidiarrheal , antifungal , antihemorrhoidal , and antioxidant agents .
saponin has also been reported for there anti - inflammatory , cardiac depressant , and hypercholesterolemic .
gc / ms chromatogram of methanol extracts of nigerian bee ( apis mellifera ) propolis shows nineteen peaks [ figure 1 ] .
the gas chromatogram shows the relative concentrations of various compounds getting eluted as a function of rt .
the heights of the peak indicate the relative concentrations of the components present in the sample .
the mass spectrometer analyzes the compounds eluted at different times to identify the nature and structure of the compounds .
the large compound fragments into small compounds are giving rise to the appearance of peaks at different m / z ratios .
these mass spectra are fingerprint of that compound which can be identified from the data library . when the mass spectra of the constituents from the propolis were compared with the nist library , a total of 60 different chemical compounds were characterized and identified .
the first compounds identified with less rt ( 13.33s ) were methyl tetradecanoate , tridecanoic acid , methyl ester , decanoic acid , methyl ester while squalene , all - trans - squalene , 2,6,10-dodecatrien-1-ol , 3,7,11-trimethyl- , ( e , e)- and farnesol isomer a [ figure 2 ] took longest rt ( 23.647s ) to identify .
methyl 14-methylpentadecanoate , hexadecanoic acid methyl ester , methyl isoheptadecanoate , and methyl tridecanoate were the most concentrated constituent as revealed by there peak height ( 26.01% ) while eicosanoic acid was the least concentrated ( peak height 0.81% ) constituent of nigerian bee propolis .
the presence of these chemical principles is an indication that in methanol extract of nigeria bee propolis if properly screened could yield a drug of pharmaceutical importance .
the presence of various bioactive principles in nigerian bee propolis extract is an indication that nigerian propolis extract , if properly screened could yield a drug of pharmacological significance .
however , the isolation of individual constituents and subjecting it to biological activity will be of medical significant .
 background : propolis is a bee ( apis mellifera ) product of plant origin with varied chemical composition depending on the ecology of the botanical origin . it has been reported in literature to possess various therapeutic effects both traditionally , clinical trial , and animal study.objectives:in the present study bioactive principle in methanol extract of nigerian bee ( a. mellifera ) propolis was determined by gas chromatography - mass spectrometry ( gc / ms ) study.materials and methods : the methanol extract of nigerian bee ( a. mellifera ) propolis was characterized for its chemical composition by preliminary phytochemicals screening and gc / ms analysis using standard procedures and methods.results:phytochemical screening revealed the presence of flavonoids , saponins , alkaloids , tannins , cardiac glycosides , anthraquinones phlobatannins , and steroids while gc / ms chromatogram revealed nineteen peaks representing 60 different chemical compounds . 
 the first compounds identified with less retention time ( rt ) ( 13.33s ) were methyl tetradecanoate , tridecanoic acid , methyl ester , decanoic acid , methyl ester while squalene , all - trans - squalene , 2,6,10-dodecatrien-1-ol , 3,7,11-trimethyl- , ( e , e)- and farnesol isomer a took longest rt ( 23.647s ) to identify . 
 methyl 14-methylpentadecanoate , hexadecanoic acid methyl ester , methyl isoheptadecanoate , and methyl tridecanoate were the most concentrated constituent as revealed by there peak height ( 26.01% ) while eicosanoic acid was the least concentrated ( peak height 0.81% ) constituent of nigerian bee propolis.conclusion:the presence of these chemical principles is an indication that methanol extract of nigeria bee propolis , if properly screened could yield a drug of pharmaceutical importance . 
INTRODUCTION
MATERIALS AND METHODS
Collection of Bee Propolis
Preparation of Propolis Extract
Phytochemical Analysis
Gas Chromatography Mass Spectrometry (GC/MS) Analysis
Identification of the Components
RESULTS
DISCUSSIONS
CONCLUSIONS