| Indocalamus Nakai is one of very common bamboo species widely distributed in the southern area of the Yangtze River from Indocalamus of Bambusodeae Nees in Gramineae. Owing to the large long leaves, they are always used as ornamental plants and the packing materials; however, most of the leaves were not fully utilized. To reveal the chemical components, study on the relationship between the components and environmental factors and promote the development and utilization of bamboo leaves from Indocalamus Nakai; Indocalamus latifolius was chosed as material, the compounds from leaves of Indocalamus latifolius were isolated and elucidated, and the antibacterial activity of some compounds were tested. The polysaccharides of contents, monosaccharide composition and antioxidant activity from leaves of Indocalamus Nakai were determined, and thermal stability and the structure of polysaccharides were studied by FT-IR, NMR and TGA. Components such as flavonoids, coumarins, aromas, elements etc. in ten species of bamboo leaves in Indocalamus Nakai were analysed and compared. The main results were as following:1. Extraction from Indocalamus latifolius leaves with95%ethanol was extracted by decoction method and purified by AB-8macroporous absorption resin. Then10%,30%,50%,70%and100%ethanol eluted respectively. The elutions were isolated by Sephadex LH-20, GX-281High Voltage HPLC Separate system and Sepacore low-pressure HPLC Separate system.25compounds were isolated from Indocalamus latifolius and5compounds were identified by comparing with standards by HPLC. On the basis of physic-chemical characters and spectroscopic analysis such as UVIR, NMR and MS,30compounds were identified as:4,4’,9’-trihydroxyl-3,3’,5,5’-tetramethoxy-7,7’-monoepoxylignan-9-O-glucoside (1); apigenin-8-C-xylose-6-C-glucopyranoside (2);4,4’,9’-trihydroxyl-3,3’-dimethoxy-7,7’-monoepoxylignan-9-O-glucoside (3); bamboo benzyl alcohol glucopyranoside I (4); bamboo lignin-7-O-glucopyranoside I (5); bamboo lignin-7-O- glucopyranoside Ⅱ(6)；bamboo benzyl alcohol glucopyranoside Ⅱ(7)；bamboo sesquiterpene (8)；phenylalanine(9)；3,5,-dimethoxy-4-benzylpropylene-phenylpropano-7-O-glucoside(10)；apigenin-2’-C-rhamnoside-6-C-glucopyranoside(11)；3"-methoxy-4-hydroxyl-phenylpropano一3,8",7",2"-cyclic-tricin-7-O-glucoside(12)；5,7,3’-trihydroxy-6-C-β-D-digitoxopyranosyl-4’-O-β-D-glucopyranosyl flavonoside(13)；apigenin-6-C-xylose-8-O-glucopyranoside(14)；3,3’,5,5’-tetramethoxy-4-hydroxyl-8-O-cinnamaldehyde-phenylpropano-7-O-glucoside (15)；3,3’,5,5’一tetramethoxy-4-hydroxyl-8-O-cinnamyl alcohol-phenylpropano-7-O-glucoside(16)；3,5-dimethoxy-4,4’-dihydroxyl-9-O-benzylacrylicester-phenylpropano-7-O-glucoside(17)；tricin-4’-O-β-guaiacylglyceryl-ether-7-O-glucoside(18)；4,4’,9,9’-tetrahydroxyl-3,3’,5,5’-tetramethoxy-7,7’-monoepoxylignan(19)；Loliolide(20)；apigenin-7-O一glucoside-6"-C-xyloside(21)；p-hydroxybenzaldehyde(22)；tricin(23)；vitexin(24)；isovitexin(25).5flavonoids were identified by comparing with standards by HPLC.They were orientin(26)；tricin-7-O-glucoside(27)；tricin-4’-O-glucoside(28)；apigenin-7-O-glucopyranoside(29)and apigenin-6-C-boivinose-7-O-glucopyranoside(30).2.All compounds were the first isolated from the leaves of Indocalamus latifolius,and the compound4,7,8,12and17were the novel.The antibacterial activities against Staphyloccocus aureus,Escherichia coli,Bacillus thuringiensis and Pseudomonas and Pseudomonas solanacearum of compounds1-10were evaluated.The results showed that the compound2and4had antibacterial activities against all the bacterium at24h in the concentration of20mg/mL and the inhibition zone were all around7mm.the inhibition zone of compound1,3and5against all bacterium were all below6.3mm.Theinhibition zone of compound6and8against Staphyloccocus aureus were6.73mm and6.57mm,respectively.3.Gas chromatography-mass spectrometry(GC-MS)was used to determine aroma components in essential oils of ten species of bamboo leaves from Indocalamus Nakai,which were obtained by Hydro-distillation.Results showed that there were twelve components such as (E)-2-Hexenal, Benzaldehyde, Heptadienal, Methyl salicylate, Safranal,(E)-Damascenone, β-ionone, Peach aldehyde, Hexahydrofarnesyl acetone, Farnesyl acetone, Isophytol and Phytol in all of essential oils. There were obvious differences in both components and relative contents among the essential oils of ten species of bamboo leaves from Indocalamus Nakai, but Aldehydes, Ketones and Alcohols were the main components.4. The content of polysaccharides from ten species of bamboo leaves in Indocalamus Nakai was determined by phenol-sulfuric acid method in the paper. The average content of total was0.43%, the range of the content was0.362-0.505%; the most content of that was I.latifolius,0.505%and the lowest of that was I.longiauritusTIANMU,0.362%.The monosaccharide composition of polysaccharides from ten species of bamboo leaves was studied by using high performance ion chromatography (HPIC) respectively. The results indicated that all polysaccharides were composed of arabinose, galactose, glucose, xylose and mannose, which content was18.341%-41.919%,20.335%-38.275%,10.660%-46.515%,5.383%-12.115%and3.244%-6.426%, respectively. The glucose content in polysaccharides of I.latifolius and I.longiauritus CHANGJIE was46.515%and40.652%, respectively; the arabinose and galactose content in polysaccharides of I.barbatus was higher than that in other polysaccharides, which was41.919%and38.275%, respectively.The antioxidant activities of polysaccharides were evaluated by DPPH and FRAP method. It indicated that all crude polysaccharides had potent antioxidant activities at the concentration of1mg/mL.The DPPH free radical scavenging ability of polysaccharide from I.pumiliswas above50%; that from I.herklotsii and I.latifolius were almost equal at35%.The thermal stability and the structure of polysaccharides from I.longiauritus CHANGJIE leaves were studied by FT-IR, NMR and TGA. The results showed that weights mean molecular weight and number mean molecular weight was43980Da,28310Da, respectively. The data of FT-IR and NMR was unanimous in suggesting the specific absorption peak of polysaccharide. Pyrolysis process of crude polysaccharide and purified polysaccharide was the same, it contained four stages. The faster speed of degradation happened at second stage (220-300℃), which the loss was50%approximately. When the temperature was above400℃, the loss was very low; As the he temperature continued to rise, it would be carbonized eventually.5. Determination of components such as flavonoids, coumarins in ten species of bamboo leaves from Indocalamus Nakai(1) The results suggested that there were significant differences on the contents of four flavonoids among the bamboo leaves. Orientin, isoorientin and vitexin were found in the leaves of I.latifolius, which content was15.56mg·kg-1,38.47mg·kg-1and2.68mg·kg-1, respectively. Orientin and isoorientin were detected in the leaves of Indocalamus Nakai except I.pedalis and I.pumilis. The content of vitexin and isovitexin was lower than that of Orientin and isoorientin in the leaves. However, the highest content of isovitexin was130.4mg·kg-1in the leaves of I.tessellatus. From the total contents of four flavonoids, the content range of was15.0-200.63mg·kg-1. The highest content was200.63mg·kg-1in the leaves of I.tessellatus; the lowest of that was15.0mg·kg-1in the leaves of I.pumilis.(2) Three kinds of coumarins were detected in the ten species of bamboo leaves, which were scopoletin,6,7-dimethoxy-coumarin and coumarin. The content range of them was7.64-89.34mg·kg-1. Scopoletin was found in all of bamboo leaves, and its content was in the range of21.99-89.34mg·kg-1;6,7-dimethoxy-coumarin was only found in the leaves of I.longiauritus TM,7.64mg·kg-1; coumarin was detected in bamboo leaves of I.tessellatus,I.latifolius, I.hirsutissimus, I.barbatus and I.longiauritus TM; the contents was in the range of7.98-11.35mg-kg’1.6. Analysis on the proteins and chlorophyll in ten species of bamboo leaves from Indocalamus Nakai(1) The range of protein content was11.70%-15.94%among the ten species of bamboo leaves from Indocalamus Nakai. The protein content in the leaves of I.latifolius; the highest content of protein was15.94%from the leaves of I.pedalis, the lowest of that was11.70%from the leaves of I.longiauritus CHANGJIE. (2) The content range of chlorophyll a, b and total chlorophyll in the ten species of bamboo leaves was1.200-2.814mg·g-1,0.602-1.250mg·g-1and1.797-3.972mg·g-1, respectively. The content of chlorophyll a, b and total chlorophyll in the leaves of I.latifoliuswas1.283mg·g-1,0.729mg·g-1and2.012mg·g-1. In other nine species of bamboo leaves, the lowest content of them was I.herklotsii, which was1.200mg·g-1,0.602mg·g-1and1.797mg·g-1, respectively. The highest content of them was I.pedalis, which was2.814mg·g-1,1.158mg·g-1and3.972mg·g-1, respectively.7. ICP-MS was applied to determine the content of17mineral elements in the bamboo leaves of Indocalamus Nakai. The trends of large amount elements in descending order were K, Ca, Mg, Mn, Fe, Al, Na, Zn, and Cu. The contents of mineral elements were low values. The average content of K,Ca, Mg and Mn were11124mg·kg-1,5549.8mg·kg-1,1241.4mg·kg-1and989.84mg·kg-1respectively. However, that of Fe was217.4mg·kg-1. The elements of Ag and Cd were not detected and the heavy metal elements of Cu, As, Hg, showed relatively low values. From the content of overall17elements in the10samples, there were higher content of elements in leaves of I.herklotsii McClure and that of I.latifolius were lower. The correlation analysis results of the elements showed thatK and Ca, Na; Ca and Mg had significantly negative correlation.Fe and A1had positive correlation among other elements except Hg. Mo had similar correlation to Fe and Al except Ca. The Hierarchical Cluster showed ten species of bamboo were classified to three classes, that were class1(I.latifolius, I.longiauritus CHANGJIE and I.herklotsii), class2(I.barbatus, I.tessellatusand I.pumilis), class3(I.hirsutissimus, I.decorus, I.longiauritus TIANMU and I.pedalis); when the distance was5-15. There were no differences among the leaves of Indocalamus Nakai, when the distance was above20. |
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