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27The Research on the Chemical Constituents of Celosia cristata L.
CHENG Da-zhong1,ZHANG Hai-Jing1*, JIN-Lan1**
3.Research Institute of Traditional Chinese Medicine, Heilongjiang University of Chinese Medicine, Harbin 150040, China
Abstract: Objective :To investigate the chemical constituents in the Celosia cristata L. Methods: The constituents of the n-Butanol portion in 60% ethanol extract are isolated and purified by means of chromatography. Structures are identified by their physicochemical characteristics andspectral features. Results:Four compounds are isolated and identified as 4-O-P-D-apifuranosyl-(1^-2)-P-D-glucopyranosyl-2-hydroxy-6-methoxyacetophenone(1),p-coumaric-acid glucoside(2) eugenyl-O-P-apiofuranosyl-(1^-6)-O-P-glucopyranoside(3),isorhamnetin3-O-(2-O-P-D-gluco-pyranosyl)-P-D-galactopyranoside-7-O-P-D-glucopyranoside(4).Conclusion: Compound 1, 2, 3and 4 are isolated from the plants of Celosia cristata L.for the first time. Compound 2, 3 and 4 are isolated from Celosia genus for the first time.
Keyword: Amaranthaceae; Celosia cristata L.; chemical constituents
Celosia cristata L.also known as chicken bun flower ,old to red, Cock flowers, Amaranthaceae in seeds is an annual herb, native to tropical America, India and Africa [1]. Celosia cristata L. inflorescence first used as medicine that contained in the "Southern Yunnan Materia Medica", sweet in flavor, cold in nature,convergingto liver and large intestine channel.It has the effect of constringency, hemostasia ,stoppingleucorrhoea and dysentery[2]. The active ingredients are flavonoids, saponins and other compounds [3-4].The compoundspossessimmunomodulatory, promoting bone growth, anti-Trichomonas vaginalisand bleeding, protectingliver and other anti-aging function[5-1 ]. Studies on chemical constituents of Celosia cristata L. few reports, I seeCelosia cristata L. seeds isolated triterpenoid saponins [4-5]whichreported in the literature.
To clarify medicinal value of Celosia cristata L. , to explore the physiological activity of the plant and material base of activeingredient,we conduct a preliminary study on chemical compositionof Celosia cristata L. inflorescence ,providing a scientific basisfor the development and utilization of Celosia cristata L. The experimental study ,12 compounds are isolated from the initial compounds which the four structures are confirmed and identified, including a phenolic glycosides 4-O-P-D-apiofuranosyl - (1 ^ 2) - P-D-glucose -2 - hydroxy-6 - methoxy-acetophenone (1);two Phenyl-propanoids:p-coumaric acid glucoside(2) and eugenyl-O-P-apiofuranosyl-(1^6)-O-P-glucoside(3);a flavonoids isorhamnetin-3-O-(2-O-P-D-glucose)-P-D-galactose-7-O-P-D-glucoside (4).
1 Materials and methods
1.1Materials
Celosia cristata L. are picked from Heilongjiang University of Chinese Medicine Medicinal Botanical Garden, and identifiedas Amaranthaceae genus Celosia cristata L. seeds byProfessor Sun Huifeng who from Department of Pharmacognosy, School of Pharmacy,Heilongjiang University of Chinese Medicine.Specimen (20110912) are kept at Research Institute of Traditional Chinese Medicine, Heilongjiang University of Chinese Medicine.
1.2 Instrumen
Bruker-400 type superconducting NMR spectrometer (USA Bruker Corporation), Agilent 1100 preparative HPLC (USA AgilentCorporation,G1361A pump, G1315B-type UV-vis detector), Agilent 1100 Series LC ChemStation workstation, C18 reverse-phase chromatographic column (ZORBAX SB-C18, 9.4 mm x 25 cm, 5 pm); FTIR7600 type Fourier transform infrared spectroscopy (Tianjin TianGuang Optical Instruments Co Ltd ); column chromatography on silica gel (Qingdao Marine Chemical, 140 to 200, 200 to 300 mesh); TLC silica gel plate (Silica ge l60 F254) and TLC Silica gel60 Rp-18 F254s (Rp-18) from the German E. Merck; reversed-phase
column chromatography ODS (ODS-AM) which is the Japanese YMC products; methanol, ethanol, petroleum ether, dichloromethane, ethyl acetate, n-butanol and other reagents were analytical grade.
2 Extraction and separation
Celosiacristata L.inflorescence 10kg, cut into small pieces, immersed in 60% ethanol at room temperature,followed by heating circumfluence for 3 times and 3hper time. The extracts are combined and suspendedin distilled water after the solvent isrecycled under reduced pressure.Extraction of suspension by petroleum ether, dichloromethane, ethyl acetate and n-butanol to a colorless solution ,the extract solvent recovery,respectively. N-butanol fraction296g.
N-butanol fractiondissolved in an appropriate amount of water and be adsorbed with a column of AB-8macroporous resin. Washed with water, 30% ethanol, 50% ethanol and 95% ethanol gradient elution,to drythe eluate fraction with reduced pressure and concentration .The part of30% ethanol is37.5g.
30% ethanol (37.5g)part ofCelosia cristata L. butanol is chromato-graphed on a silica gel column extracted by column chromatography on silica gel ,and washed with dichloromethane -methanol (20:1 to 1:1) gradient elutionto give six components (F 1 ~ F 6). Part F 3 (1.5g) by normal phase silica gel column chromatography, and recrystallized with methanol to give compound 2 (4.3mg). Part F 4 (6.8 g) by ODS column chromatography HPLC (preparative) is isolated the compound 1 (5.0mg), compound 3 (8mg). Part F 5 (8.7 g) by ODS column chromatography and HPLC (preparative) isolated the compound 4 (7.2mg).
3 Results and discussion
Compound 1: white amorphous powder, soluble in methanol. Ferric chloride color reaction is positive, suggesting that themolecular structure may contain phenolic hydroxyl groups. With 5% a-naphthol ethanol solution and concentrated sulfuric acid,Molish reaction is positive, and generated purpleringbetween thesurface, indicating the molecular structure may be sugar or glycosides.The UV spectrum of the Compound 1 in methanol solution at 210nm and 284nm ,there are two major absorption bands.
HR-ESI-MSm/z 461.1589[M+H]+,molecular formula C20H28O12, unsaturated degree 7.XH-NMR (CD3OD) 5:6.22 (1H, d, J=2.3 Hz, H-3), 6.16 (1H, d, J=2.3 Hz, H-5), 5.44 (1H, d, J=1.5Hz, H-1"), 5.05 (1H, d, J=7.3 Hz, H-1'), 4.04 (2H, d, J=9.5 Hz, H-4"), 3.92 (1H, d, J=1.5 Hz, H-2"), 3.90 (3H, s, 6-OCH3), 3.79 (2H, d, J=9.5 Hz, H-4"), 3.61 (2H, dd, J=14.0 Hz, 6.0 Hz, H-6'), 3.50 (2H, s, H-5"), 2.58 (3H, s, -CH3).
13C-NMR (CD3OD) 5 : 204.9 (C=O), 167.7 (C-2), 165.4 (C-4), 164.7 (C-6), 107.8 (C-1), 97.7 (C-3), 92.8 (C-5), 56.4 (6-OCH3), 33.2 (8-CH3), 100.1 (C-1'), 78.1 (C-2'), 78.4 (C-3'), 71.4 (C-4'), 78.5 (C-5'), 62.5 (C-6'),110.1 (C-1"), 78.5 (C-2"), 80.8 (C-3"), 75.5 (C-4"), 66.0 (C-5").The above data consistent with the dataliterature [11] reported, it is identified as4-O-P-D-apifuranosyl- (1—>2)-P-D-glucopyranosyl-2-hydroxy-6-methoxyacetophenone.
Compound 2: colorless granular-crystal, soluble in methanol. Molish reaction is positive, suggesting that the structure may be glycosides.
HR-ESI-MSm/z 327.1009[M+H]+, molecular formula C15H18O8, unsaturated degree 7.XH-NMR (CD3OD) 5 : 7.63 (1H, d, J=15. 9Hz, H-7), 7.39 (2H, d, J=8.6 Hz, H-2, 6), 6.72 (2H, d, J=8.6 Hz, H-3, 5), 6.27 (1H, d, J=15.9 Hz, H-8), 5.47 (1H, d, J=7.8 Hz, H-1'), 3.75 (1H, dd, J= 11.9 Hz, 1.6 Hz, H-6'a), 3.59 (1H, dd, J =12.0 Hz, 4.5 Hz, H-6'b).
13C -NMR (400MHz, CD3OD) 5: 167.7 (C=O), 161.6 (C-4), 148.0 (C-7), 131.4 (C-2, 6), 127.0(C-1), 116.9 (C-3, 5), 114.5 (C-8), 95.8 (C-1'), 78.8 ( C-3'), 78.0 (C-5'), 74.1 (C-2'), 71.1 (C-4'), 62.4 (C-6').The above data and the literature [12] reported are basically the same, it is identified asp-coumaric acid glucoside.
Compound 3: colorless powder, soluble in methanol. Molish reaction is positive, suggesting that the structure may be glycosides. The UV spectrum of the Compound 3 in methanol solution at 201.8nm and 276.2nm, there are two major UV absorption peak. Based on the above information, suggesting that the compound is phenylpropanoid compounds.
HR-ESI-MSm/z 443.1847[M+H]+, molecular formulaC21H30O10, unsaturated degree 7.*H-NMR(CD3OD) 5:7.07 (1H, d, J=8.2 Hz, H-6), 6.82 (1H, d, J=1.8 Hz, H-3), 6.74 (1H, dd, J=8.2 Hz, 1.8 Hz, H-5), 5.96 (1H, ddt, J=16.9 Hz, 10 Hz, 2.0Hz, H-8), 5.08 (1H, dd, J=16.9 Hz, 2.0 Hz, H-9b), 5.04 (1H, dd, J=10 Hz, 2.0 Hz, H-9a), 4.96 (2H, dd, J=2.4 Hz, 9.4 Hz,H-7), 4.95 (1H, d, J=2.4 Hz , H-1"), 4.79 (1H, d, J=7.2 Hz, H-1'), 3.99 (1H, dd, J=1.5 Hz, 11.0 Hz, H-6'), 3.73 (1H, d, J=9.6 Hz, H-4"a),3.94 (1H, d, J=9.6 Hz, H-4"b), 3.88 (1H, d, J=2.4 Hz, H-2"), 3.84 (3H, s, OCH3), 3.56 (2H, s, H-5").
13C-NMR (CD3OD) 5: 150.8 (C-2), 146.3 (C-1), 139.0 (C-8), 136.5 (C-4), 122.2 (C-5), 118.4 (C-3), 115.9 (C-9), 114.1 (C-6), 111.0 (C-1"), 103.1 (C-1'), 80.5 (C-3"), 78.0 (C-3'), 77.8 (C-5'), 77.0 (C-2"), 75.0 (C-4"), 74.9 (C-2'), 71.6 (C-4'), 68.7 (C-6'), 65.6 (C-5"), 56.7 (OCH3-2), 40.8 (C-7). The above data consistent with the dataliterature [13] reported, it is identified aseugenyl-O-P-apiofuranosyl-(1^-6)-O-P-glucopyranoside.
Compound 4: yellow crystals, soluble in methanol, dimethyl sulfoxide. TLCspot isyellow ,yellow fluorescent enhancementunder UV light after using 10% sulfuric acid - anhydrous ethanol ; Ferric chloride color reaction is positive, hydrochloric acid - Mg reaction is positive, Molish reaction is positive. The UV spectrum of the compound 4 in methanol at 253.7nm and356.1 nm ,there are two major peaks.Further speculated that the compound is flavonoids.
HR-ESI-MSm/z 771.2277[M+H]+ , molecular formulaC34H42O20, unsaturated degree 14.*H-NMR (DMSO-d6) 5: 7.94 (1H, d, J=2.0 Hz, H-2'), 7.49 (1H, dd, J= 8.4 Hz, 2.0 Hz, H-6'), 6.91 (1H, d, J=8.4 Hz, H-5'), 6.41 (1H, d, J=2.0 Hz, H-8), 6.18 (1H, d, J=2.0 Hz, H-6), 5.75 (2H, d, J=7.4 Hz, H-1", H-1""), 4.24 (1H, d, J=7.0 Hz, H-1"'), 3.86 (3H, s, OCH3), 3.58 (1H, m, H-6"a,), 3.55 (1H, m, H-6"b), 3.39 (1H, m, H-6"'), 0.74 (3H, d, J=6.2 Hz, H-6"").
13C-NMR(DMSO-d6) 5 : 177.1(C-4), 164.8(C-7), 161.1(C-5), 149.3(C-4'), 146.8(C-3'), 132.4(C-3), 121.4(C-6'), 120.4(C-1'), 115.2(C-5'), 113.4(C-2'), 105.0(C-10), 103.7(C-1"'), 100.7(C-1""), ,98.9(C-6), 98.1(C-1"), 93.8(C-8), 78.0(C-2"), 77.4(C-5""), 76.9(C-5"'), 76.8(C-3"'),76.5(C-5"), 69.9(C-4"'), 69.7(C-4""), 76.9(C-3""), 66.6(C-4"), 62.7(C-6""), 60.9(C-6"'), 60.4(C-6"), 55.6(OCH3).The above data and the literature [14] reported are basically the same,it is identified as isorhamnetin3-O-(2-O-P-D-gluco-pyranosyl)-P-D-galactopyranoside-7-O-P-D-glucopyranoside
4 Conclusions
The structure confirmation and identification, specific structure of four compounds are as follow:
Number English name Structure
OH
4-O-P-D-
apifuranosyl- (1^2)-Compound P-D-glucopyranosyl-
1 2-hydroxy-6-
methoxyacetophenon e
OHOH
Compound 2
p-coumaricacid glucoside
OH
//—COOH
ho
Compound 3
eugenyl-O-ß-
apiofuranosyl-(1^6)-
O-ß-glucopyranoside
v / I _o a
OHOH
Compound 4
isorhamnetin3-O-(2-
O-ß-D-gluco-
pyranosyl)-ß-D-
galactopyranoside-7-
O-ß-D-
glucopyranoside
oh
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Study on thePreparingProcess ofIntermediates of Anti-cancer Element as Colon
Targeting Dropping Pill
ZHANG Xiao-Yan1, Yu Ying1, XIE Ning2, MA Wei3, DONG Pei-liang1
(1Heilongjiang University of Chinese Medicine, Research Institute of Traditional Chinese Medicine, Harbin 150040,China The First Affiliated Hospital of Heilongjiang University of Chinese medicine;3)
Abstract: objectiveThe preparing technique of the droppingpill were established.Methods The orthogonal test was design with PEG4000, PEG6000 and Boluosham as base for the research of the dropping pill Preparation process. Resuits The best preparing technique were thedrug and matrix as 1:5, PEG4000/PEG6000 as 3:1, drug / Boluosham as 20:1, liquid temperature as88C, and dropping rate as 20 drops per min, and dropping distance as 5cm.Conclusions The dropping pill was prepared with the best hardness, roundness and weight differences. Key words :anti-cancer element; colon targeting; dropping pill
Keaisu is a colon-targeted preparations, including curcuminoids from the root of curcuma longa and quercetin from bud of sophora japonica. Quercetin is the hydrolysate which from rutin is hydrolyzed under the action of enzymes or acid. Modern pharmacological studies have shown thatQuercetin is a kind of natural flavonoid compound which can dilate coronary artery, reducing blood lipid, anti-inflammation, anti-allergy, anti-diabetes mellitus complication and other effects[1]. Curcumin is a yellow, acid, phenolic compound extracted from a plant of Zingiberaceae and Curcuma genus, the root of curcuma longa. And Curcumin is the major active ingredient pharmacology. Curcumin has extensive pharmacological effects, such as anti-tumor, anti-inflammatory, antioxidant, anti mutation, anti-hiv, etc[2]. The Chinese medicinetargeted preparations are composed of curcuminoids and quercetin (mass ratio35g: 20g) [3], which are effective components, and thiswill provide a reference for studyingon oral drug release system.
1. Material
Quercetin (Xian xiaocao zhiwukeji Co., LTD., UV purity is 98.05%); Curcuminoids (Ningbo chengtai modern Chinese medicine technology Co., LTD, total contents of curcuminoids by HPLC is 95.0%); PEG4000 * 6000 (Tianjin kemiou); dropping pill preparation device (homemade).
2. Method and Results[4] 2.1 Dropping distance
1FundHeilongjiang University of Chinese Medicine Doctor Innovation FundNOB201006
Author ProfileZHANG Xiao Yan, female, associate professor, doctor major in Chinese medicine, mainly engaged in material base of traditional Chinese medicine and novel drug delivery system. Tel045187266836.
E-mail: [email protected].
