井冈霉素的提纯和含量测定研究英文

井冈霉素的提纯和含量测定研究英文 * CAO Hui，WANG Shao-yun School of Environment and Biology，Kaili Institute，Kaili 556011，China Abstract ,Objective, In order to get the standard substancJingge oanfg mycin usedin quality control，Jinggangmycin wasp urified fromi ts original pesticide and its content wadest ermined, ,Method, Mixed solvents weres elected to recrystallize Jinggangmycin ( content6 2% ) ,P hen ol-sulfuric acid colorimetric method aws adopted dtoet ermine its content，and the effectsiff oerfe dnt determination conditions on the contenrets ults werea lso studied, ,Result, The purity of the smaples after re-crystallization wasi ncreasde to 97, 7% ,T he o ptimal conditions for phenol-sulfuric acid colorimetric method dtoet ermine the content oJifn ggangmycin wereas follows: anhydrougsl ucose as cnotrol sample，5% phenol，1, 0 ml phenol solution，5, 0 ml sulfuric acid，stilly placed for 20 min afterr eaction，490 nm as edtec- 2 tion wavelength; the correlation coefficient of glucose standard curv(e R ) was0 , 993 9; the average recovery of standaarddd i tion was9 9, 72% ，RSD was 0, 362 8% ,, C onclusion, Re-crystallization is a good way retofin e Jinggangmycin, Phenol-sulfuric acid colorimetric method can be usededt etorm ine the content of Jinggangmycin with simple operation，high accuracy ansdtr ong reliability，which can be iwdely applied in industrial production, Key words Jinggangmycin; Re-crystallization; Phenol-sulfuric acid colorimetric method Jinggangmycin， also known as 2-, 2，3-Dihydroxy-6- terial of Jinggangmycin ( produced by WuhaJnia kailong Technolo- ( hydroxymethyl) -4-,,4，5，6-trihydroxy-3-( hydroxymethyl) -1-cy- gy Development Co, ，Ltd, ) ，ethyl acetate( analytical reagent， clohex-2-enyl,amino,cyclohexoxy,-6-( hydroxymethyl ) oxane-3， producedby Tianjin Bodi Chemical Co, ，Ltd, ) ，methanol ( ana- 4，5-triol，Validamycin A，Solacol，Validacin andV alimon，etc, ， lytical reagent，producebd y Tianjin Fuyu Fine Chemical Co, ， is composedb y six glucosamine compound(s know n aso mcbina- Ltd, ) ，glucose ( analytical reagent，produced Sbiyn opharm Chemi- tion A，B，C，D，E andE ) ，of which A，B，E andF accounto r fcal ReagenCt o, ，Ltd, ) ，phenol ( analytical reagent，produceby d ,1,about6 0% ，and the amin active ingredient is A, They aret he Tianjin DamaoC hemical Co, ，Ltd, ) ，concentratedsu lfuric acid secondary metabolites of Jinggangmycin producing strain，original ( analytical reagent，producebyd B eijing Beihua Fine Chemical actinomycete streptavidin of J inggangmycin producing strain be- Co, ，Ltd, ) ，0, 531 9 mol / L phenol solution ( home-mad) e, longs to absorbengrto up, 1, 2 Experimental methods1, 2, 1 Re-crystallization, The raw material of Jinggangmycin and a Jinggangmycin is the internal absorption fungicide，which is certain amount osfo lvent alcohol were addeidnt o the conical mainly used too cntrol rice sheath lbight with the duration of 15) flask，and the mixture was heated to make them artaewria l dis- 20 d，being able to bear reosion of rain, It also can be used toon -c solve in solvent completely, Then thes olution was ifltered into the trol Sclerotial disease，green smut，corn northelrean f blight， beakerw ith folded filter paper，and addeitdh wethyl acetateun til no sediment precipitated out, Finally，Büchner funnel was usetdo maize earr ot，damping-off and Slcerotium rolfsii in maize andl eg- filter the solution，and the filtrate was used to walsahsk ,f The fil- umes，damping-off in ginseng，cotton and vegetable, In order to ter cake waisrs tf placed in vacuum ove( n5 0 ? ) for abou1t h， obtain the productsi thw high purity and ocntrol the production then transferreidn to the brown dryecro ntaining anhydrouCs aCl2 process，the ramwat erial of Jinggangmycin needs to beu ripfied, for 30 min，and the tessta mples of Jinggangmycin waso btained, In this study，Jinggangmycin was uprified by re-crystallization 1, 2, 2 Determination principle of polysaccharides, Determination of method，and its content wasn aalyzed by phenol-sulfuric acid col- polysaccharide content iwth phenol-sulfuric acid colorimetric method ,2 ) 3,orimetric method, was raised by Dubois et al, , Polysaccharides or oligo- saccharides were hydrolyzed by concentratesdu lfuric acid at the appropriate high temperature，ando nmosaccharide was rpoduced， 1Materials and Methods then transferredn to furfura dervatves throughr apd dehydra-iliii 1, 1 Reagentsan d nstuments Man nstruments usedi n theiriitest were A L204 electronic balance ( Mettler-Toledo Group ) ， tion, The derivative had oclor reaction with phenol under strong SHZ-D circulating water type vacuum pu( mBepiji ng experi- ? acid condition，and its absorbance value A showedli near relation- mental equipment Co, ， Ltd, ) ， DZ-1BC vacuum r yding oven ship with sugar concentration C at the wvaelength of 490 nm，so ( Tianjin Theis special apparatuCso , ，Ltd, ) and TU-1900d ouble beam UV-visible spectrophotome(t eBre ijing Puxi General Instru- trol solution and tesst olution was xeactly taken and addientdo 10 ml volumetric flasks，then added iwth 0, 2，0, 4，0, 6，0. 8，0, 9， 2Results and Analysis 1, 0 and1 , 1 ml pheno souton，respectvey，and qucky drpped lliilili 2, 1 Detemination of the best detection wavelength fo poly-rr with 5, 0 ml concentratesdul furic acid，finally added iwth distilled saccharideControl solution and test so lution were s eparately water to the constant volume of 10 ml, The absorbance value A of taken foro lcor development by phenol-sulfuric acid solution, The the solution was dteermined at 490 nm，and thsuei table dosagef ofull wavelengths of boths olutions were scannebdy TU-1900d ouble pheno was dteermned, libeam UV-visible spectrophotometer within the range of 40)060 0 1, 2, 5 Selection of the dosageu lofufr isc acid, 7 copies of 1, 0 nm，and both of them hamda xthmeu m absorpton at the wvea- iiml control solution and tesst olution was xeactly taken anda dded length of 490 nmi thw the same spectra，so 490 nmsel ewctaesd as into 10 ml volumetric flasks，then separately added iwth 1, 0 ml the detection wavelength for polysaccharide, pheno souton，and qucky drpped wth 2, 0，3, 0，4, 0，4.5 ， lliilii2, 2 Selection of the dosagoe f phenol As shownin Fig, 1 and 5, 0，5, 5 and 6, 0 ml concentratesdu lfuric acid，finally added Fig, 2，when thedo sage oconf centratesdul furic acid was nesured with distilled water to the consvtanolutm e of 10 ml, The absor- to be excessive，the absorbance value was stable with the phenol bancev alue A of thes olution was dteermined at 490 nm，andt he dosage 0o,f 6 ) 1, 0 ml, When thep henol dosage wa1,s 1 ml，the suitable dosage oufl fusric acid was dteermined, absorbance value was derceased, Therefore，0, 1 ml phenol was 1, 2, 6 Selection of reaction time, Five copies of 1, 0 ml control the best for thete rdmeination of the oplysaccharide content, solution and tesst olution was xeactly taken and addientdo 10 ml volumetric flasks，then added iwth 1, 0 ml phenol solution，and quickly dripped with 5, 0 ml concentratesdul furic acid，respec- tively, The reaction times were 15，20，25，30 a3n5d m in, The absorbance value A of thes olution was dteermined at 490 nm，and the time with stable color development was selected to determine the absorbance value, 1, 2, 7 Preparation of standarsdo lution, 0, 015 2 g glucose was accurately weighed，and placed in 100 ml flask, Then watewr as added toi ssdolve the glucose with the total volume of 100m l, The solution was veenly mixed，and the standarsdol ution of lgucose ) 6 was successfully prepared itwh the concentration of 0, 84 × 1 0 mol / ml, Fig, 1Effect of phenol dosagein glucose on absorbance value1, 2, 8 Preparation of standardu rvce, According to the ocnfirmed conditions for dteermination，0, 1，0, 2，0, 3，0, 4，0, 5，0, 6， 0. 7，0, 8，0, 9 and 1, 0 ml glucose standards olution was xeactly taken andp laced in 100 ml flask, Then thes olutions were added with 1, 0 ml phenol solution，respectively, After evenly mixed， the solutions were uqickly dripped with 5, 0 ml concentratesdul fu- ric acid，and added iwth distilled water to the consvtanolutm e of 100 ml，mixed, The absorbance value A of thes olution was dteer- mined at 490 nm by spectrophotometer, The standard curvbee- tweenA value and ocncentration C was rdawn, Linear regression was crraied out，and thee quation of standard curve owabtas ined, 1, 2, 9 Recoveryt est, Three copies of 1.0 ml test solution was Effect of phenol dosagein Jinggangmycin on absor-Fig, 2exacty taken，and detected throhuegnoh -spufuric acid coori- llll bance valuemetrc method aoccrdng to thes eected condtons, The concen- iilii tration of polysaccharides was claculated according to standard 2, 3 Selection of the dosageo f sulfuric acid As showni ncurve，and the meavanlu e was laso calculated, Then certain vol- Fig, 3 and Fig, 4，when theph enol dosage wa1,s 0 ml，with the ume ogf lucose standards olution was addeidn to the solutions，re- increase of concentratesudl furic acid，the absorbance value was spectively, The polysaccharide content was detected throughe-h p Fig, 6 Effect of reaction time on absorbance value of Jing- gangmycin solution Fig, 3 Effect of sulfuric acid dosagein glucose on absorbanceCV? 0 1) 8 value When V= 0, 1 ml，C== 0, 84 × 10 mol / ml 1 1 V Similarly，the corresponding glucose concentrations as V = 0, 2，0, 3，, , ,1 , 0 m l ,we r e ,cla c ula,te d, Through theli near regression of data，three gression equation betweeno nccentration and asborbance value during the determina- tion process of polysaccharides waso btained as follows: A = 0, 065 3C +0, 039 3( 1) In the formula，A was asborbance value，C wasg lucose con- centration ( mol / ml) ,2 Correlation coefficient R= 0, 993 9， indicating that the equation had goodness iot,f f Fig, 4 Effect of sulfuric acid dosagein Jinggangmycino n ab- 0, 043 2 g Jinggangmycin sample was accurately weighed and sorbance value dissolved in water，then thseo lution was transferreid nto 100 ml 2, 4Reaction time and stabilityAs showni n Fig, 5 andflask，and added iwth water to the consvtanolutm e，mixed, 0, 5 ml solution was transferreindt o 10 ml flask by pipet，adde dwith Fig. 6，with the prolongation of p lacement time，the absorbance value was gradually increased，and the variation was tend to be 1, 0 ml phenol，and quickly dripped with 5, 0 ml concentratesudl - furic acid, After reaction for 20 min，the absorbance value was stable after thes olution was lpaced for 20 min, Therefore，2m0 in wass elected as the appropriate reaction time for thete st, measured at thev ewleangth of 490 nm，and= 0A , 316 8, When A =0, 316 8 was s ubstituted into formula ( 1 ) ， ) 8 C= 4, 25 × 10mol / ml waso btained, determination So the inJggangmycin content wsa C× 10 ×M × 100 determination W =× 100% = 97, 7% 0, 5 × 0, 043 2 M in the ofrmula represented tmheol ecular weight of Jinggangmycin,2, 6 Recovery of standard addition test With glucose stand- ard as theo nctrol，the Jinggangmycin content wasn aalyzed by sul- furic acid-phenol method，the recovery of standaitirodn waddas 99, 72% ，and RSD was, 3602 8% ，indicating that the method had high degree of accurac(y T able 2) , Fig, 5 Effect of reaction time on absorbance value of glucosesolution 2, 5 Pepaaton of standad cuve Based on the abovnae- a rrirrl iscussionD3yss，the best ocndton for the tdeermnaton of Jnggangmycn iiiiiiiThe poysaccharde detecton prncpe of pheno-sufurc acd liiiillliithrough hpenol-sulfuric acid method was aosll ofws: 5% phenol， methods as foows: poysaccharde compounds arers t fhydro- illlii ,6, the reaction time during the measuremernotc epss , method depends ont rtahnesi ent high temperature asntrdo ng acidThe results showed that tmheix ed solvent usedi n the test ws a environment producedb y the action of concentratesudlf uric acid， advisable，which can ahcieve the purpose oufr ifpication，and the Table 1 Determination results of recovery of standard addition purity could be increased from abou6t2 % to 97, 7% ,U V sp ec- Additiontrum has been used fouar ntqitative analysis of J inggangmycin， dosagePolysaccha- verageStandardSDAR ride quantity of glucose MeasuredecoveryRwhich is fast ands imple with easy claculation, But it needst o deviationrecovery %No, in samples standard valuerate?% rate%%? strcty and accuratey wegh the quantty of sampe，and the accu- illiilsamole mg mgracy orfe sults mainly depends on trheep roducibility of sample size 99, 720, 351 8 0, 362 810, 068 530, 010, 078 5199, 80and the stability of operation conditions, 2 0, 068 53 0, 02 0, 088 60 100, 30 3 0, 068 53 0, 03 0, 098 32 99, 30 The besto ncdition for the dteection of Jinggangmycin by phe- 4 0, 068 53 0, 04 0, 108 28 99, 38 no-sufurc acd coormetrc method has beeonn frcmed through lliiliii 5 0, 068 53 0, 05 0, 118 47 99, 88 comparison test: anhydrougslu cose asc ontrol，5% phenol，1, 0 ml phenol solution，5, 0 ml sulfuric acid，stilly placed for 20 min after reaction，detection wavelength 490 nm; the correlation coefficient of 2 glucose standard curv( eR ) was0 , 993 9, The detection of Jing- gangmycin content under the boensdti ticon has simple operation， high accuracy anstdro ng reliability, The methodis simple and fsat without requirement of polysaccharide pureness andxp eensive con- trol equipment，which can be used tote rdmeine the water-soluble and insoluble samples, References ,1, CHEN L，JIANG GP，PAN CP, Determination of iJngangmycin A by re- ig, 7 Standard curve of glucoseFversed-phase HPLC,J,, Agrochemicals，2007，46 ( 10) : 686 )68 7, ( in Chinese) , thus making Jinggangmycin quickly hydrolyze, Therefore，thceo n- ,2, DUBOIS M，GILLES KA，HAMILTON JK，et al, Colorimetricmethod for determination of sugars andrelated substances,J,, Anal Chem, 1956，28: centratedsu lfuric acid should be quickly dripped andi ntensely hit 350 ) 351, ( in Chinese) , the liquid surface to productrae ns ient high temperature， this ,3, DUBOIS M，GILLES KA，HAMILTON JK，et al, A colorimetric methodo fr would be conducive to the ocmplete hydrolysis of Jinggangmycin， the determination of sugars,J,，Nature, 1951，168: 167 ) 169, and thesl ow dripping of uslfuric acid will lead to the error oxf- e,4, ZHANG Q，ZHANG TM, Determination of polysaccharide contentb y phe- nol-vitriolic colorimetry,J,, Shandong Foodie ncSec and Technology，2004 perimental result, Phenol is easy to boex idized whenit is exposed in ( 7) : 46 ) 50, ( in Chinese) , air，forming red compound, Therefore，thed etection process ,5,MAO Y, Optimum scheme andu rpity appraisal of extracting soluble sugar in should be rapid in order to vaoid the impact of phenol oxidation on pl, amarusl eaves,J,, Journal of BamboRoesea rch，2002，2( 13) : 46 ) 50, ( in Chinese) , determination results, During the formation process of f urfural， ,6, ZHOU YB，WANG W，ZHOU XR, Determination of polysaccharides in five-carbon aldose and methyl pentose ldaose are usually easier Phyllostachys pubescenlesa ves by phenol-vitriolic colorimetry,J,, Modern than six-carbon aldose，and the producist also Food Siecnce , Technology，2008，24( 3) : 180 ) 181, ( in Chinese) , 井冈霉素的提纯和含量测定研究 曹 晖，王绍云 ( 凯里学院环境与生命科学学院，贵州凯里 556011) 摘要 ,目的,对井冈霉素原药进行提纯，并测定其含量，以用于井冈霉素生产中的质量控制。,方法,选用混合溶剂对 62% 井冈霉素原药进行 重结晶; 利用苯酚硫酸比色法对其含量进行测定，并研究了不同测定条件对含量测定结果的影响。,结果,经过重结晶后的样品，其纯度提高 " 至 97, 7% 。苯酚硫酸比色法测定井冈霉素含量的最佳条件为: 对照品为无水葡萄糖; 苯酚用量 5% ; 苯酚溶液 1, 0 ml，浓硫酸用量 5, 0 ml，反应 "2 后静置 20 min，检测波长 490 nm; 葡萄糖标准曲线的相关系数( R) 为 0, 993 9; 平均加标回收率为 99, 72% ，RSD 为 0, 362 8%。,结论,重结晶法 可以很好的完成井冈霉素的精制; 硫酸苯酚比色法用于测定井冈霉素的含量，操作简单，准确度高，可靠性强，可以在工业化生产中推广应用。 " 关键词 井冈霉素; 重结晶; 硫酸苯酚比色法 " file:///C|/Users/Administrator/Desktop/新建文本文档.txt 涵盖各行业最丰富完备的资料文献,最前瞻权威的行业动态,是专业人士的不二选择。 file:///C|/Users/Administrator/Desktop/新建文本文档.txt2012/8/26 12:19:58