MD585Z - Method for quantitative determination of flavonoids in egetable raw material - Google Patents
Method for quantitative determination of flavonoids in egetable raw material Download PDFInfo
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- MD585Z MD585Z MDS20120065A MDS20120065A MD585Z MD 585 Z MD585 Z MD 585Z MD S20120065 A MDS20120065 A MD S20120065A MD S20120065 A MDS20120065 A MD S20120065A MD 585 Z MD585 Z MD 585Z
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- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 43
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 42
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 42
- 239000002994 raw material Substances 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 16
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 claims abstract description 48
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 40
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 24
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000000284 extract Substances 0.000 claims abstract description 20
- 235000005875 quercetin Nutrition 0.000 claims abstract description 12
- 230000003287 optical effect Effects 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 239000007790 solid phase Substances 0.000 claims abstract description 8
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 claims description 11
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 claims description 11
- 238000010521 absorption reaction Methods 0.000 claims description 11
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims description 11
- 229960001285 quercetin Drugs 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 10
- 238000010790 dilution Methods 0.000 claims description 4
- 239000012895 dilution Substances 0.000 claims description 4
- 239000012675 alcoholic extract Substances 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 abstract description 3
- 235000013311 vegetables Nutrition 0.000 abstract 2
- 238000002481 ethanol extraction Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 11
- 235000019441 ethanol Nutrition 0.000 description 11
- IKGXIBQEEMLURG-NVPNHPEKSA-N rutin Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-NVPNHPEKSA-N 0.000 description 8
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 7
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 7
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 7
- 235000005493 rutin Nutrition 0.000 description 7
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 7
- 229960004555 rutoside Drugs 0.000 description 7
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 5
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 5
- 229930002875 chlorophyll Natural products 0.000 description 5
- 235000019804 chlorophyll Nutrition 0.000 description 5
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 5
- 238000010992 reflux Methods 0.000 description 5
- 238000009835 boiling Methods 0.000 description 4
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 235000017309 Hypericum perforatum Nutrition 0.000 description 3
- 244000141009 Hypericum perforatum Species 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000005375 photometry Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 229930000044 secondary metabolite Natural products 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- GMGIWEZSKCNYSW-UHFFFAOYSA-N 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one;dihydrate Chemical compound O.O.C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 GMGIWEZSKCNYSW-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000011197 physicochemical method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 150000003243 quercetin Chemical class 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- -1 trihydrate rutin complexes Chemical class 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
Description
Invenţia se referă la domeniul farmaceutic, în special la un procedeu de determinare cantitativă a flavonoizilor în materia primă vegetală. The invention relates to the pharmaceutical field, in particular to a process for quantitative determination of flavonoids in plant raw material.
Este cunoscut procedeul de determinare cantitativă a flavonoizilor în materia primă vegetală pe baza schimbării densităţii optice la maximumul de absorbţie al complecşilor de flavonoizi cu clorură de aluminiu (AlCl3). În acest scop se propune fierberea cantităţii exacte de materie vegetală timp de 20…30 min cu etanol 30…70% în raport de 1:(100…200) într-un balon cu un răcitor cu reflux, după care se efectuează răcirea, se aduce cu etanol corespunzător la masa iniţială, proba alicvotă filtrată a extractului se tratează cu soluţie de clorură de aluminiu (AlCl3) în etanol 70% şi se măsoară densitatea optică a soluţiei la lungimea de undă analitică de 410…415 nm la un spectrofotometru pe fonul probei care nu este tratată cu clorură de aluminiu şi este acidulată cu acid acetic. În paralel, în aceleaşi condiţii se efectuează experienţa şi cu proba cu 1,5 ml de soluţie de rutină 0,1%. Conţinutul de flavonoizi se calculează în unităţi de rutină [1]. The procedure for quantitative determination of flavonoids in plant raw material is known based on the change in optical density at the absorption maximum of flavonoid complexes with aluminum chloride (AlCl3). For this purpose, it is proposed to boil the exact amount of plant material for 20…30 min with 30…70% ethanol in a ratio of 1:(100…200) in a flask with a reflux condenser, after which it is cooled, brought to the initial mass with the appropriate ethanol, the filtered aliquot sample of the extract is treated with a solution of aluminum chloride (AlCl3) in 70% ethanol and the optical density of the solution is measured at the analytical wavelength of 410…415 nm with a spectrophotometer against the background of the sample that is not treated with aluminum chloride and is acidified with acetic acid. In parallel, under the same conditions, the experiment is also performed with the sample containing 1.5 ml of 0.1% rutin solution. The flavonoid content is calculated in rutin units [1].
Dezavantajele principale ale procedeului cunoscut sunt: la determinare se presupune că numai flavonoizii (FL) formează un complex cu AlCl3, nu se ia în consideraţie efectul de denaturare asupra rezultatelor analizei de formare a complexului compuşilor secundari cu AlCl3. În primul rând se denaturează rezultatele analizei din cauza prezenţei clorofilei extrase din plantă cu alcool etilic. Clorofila în regiunea 350…450 nm are aşa-numitele benzi de absorbţie Sore, intensitatea de absorbţie a cărora scade după acţiunea cu AlCl3 care afectează rezultatele analizei. Determinarea în mai multe etape duce la o creştere a timpului analizei şi include obţinerea extractului, luarea de probă alicvotă, prepararea soluţiei de lucru şi a soluţiei de referinţă cu şi fără adaos de AlCl3, prepararea soluţiei standard de rutină, de asemenea, cu şi fără adaos de AlCl3. Rezultatele analizei se exprimă în unităţi de rutină, dar dacă materialul analizat conţine FL, alţii decât de rutină, este necesară recalcularea luând în consideraţie diferenţa dintre masele lor moleculare. Deci, în dependenţă de raportul probei de rutină : quercetină se observă o schimbare în intensitatea de absorbţie a luminii, diferenţa dintre absorbţia de rutină şi quercetină creşte la deplasarea absorbţiei totale la lungimi de undă mai lungi, ceea ce duce la o denaturare foarte puternică a rezultatelor de determinare a FL. Dacă măsurile fotometrice se efectuează în intervalul 410…415 nm, atunci rezultatele analizei depind foarte mult de compoziţia flavonoizilor extractului. The main disadvantages of the known procedure are: when determining it is assumed that only flavonoids (FL) form a complex with AlCl3, the denaturing effect on the analysis results of the formation of the complex of secondary compounds with AlCl3 is not taken into account. First of all, the analysis results are denatured due to the presence of chlorophyll extracted from the plant with ethyl alcohol. Chlorophyll in the region of 350…450 nm has the so-called Sore absorption bands, the absorption intensity of which decreases after the action of AlCl3, which affects the analysis results. Multi-stage determination leads to an increase in the analysis time and includes obtaining the extract, taking an aliquot sample, preparing the working solution and the reference solution with and without the addition of AlCl3, preparing the routine standard solution, also with and without the addition of AlCl3. The analysis results are expressed in routine units, but if the analyzed material contains FL, other than routine, it is necessary to recalculate taking into account the difference between their molecular masses. So, depending on the ratio of routine sample: quercetin, a change in the light absorption intensity is observed, the difference between routine and quercetin absorption increases when the total absorption shifts to longer wavelengths, which leads to a very strong distortion of the FL determination results. If photometric measurements are carried out in the range of 410…415 nm, then the analysis results depend greatly on the flavonoid composition of the extract.
Problema pe care o soluţionează invenţia constă în elaborarea unui procedeu de determinare cantitativă a flavonoizilor în materia primă vegetală cu un număr mai mic de operaţii luând în consideraţie impactul asupra rezultatelor de determinare şi a compuşilor extractibili secundari. The problem solved by the invention consists in developing a process for quantitative determination of flavonoids in plant raw material with a smaller number of operations, taking into account the impact on the determination results and secondary extractable compounds.
Conform procedeului revendicat, se efectuează extragerea materiei prime vegetale cu etanol de 70…75%, tratarea cu clorură de aluminiu, determinarea fotometrică a densităţii optice a extractului şi calcularea cantităţii de flavonoizi, iar înainte de extragerea cu etanol materia primă se extrage cu cloroform, după care cloroformul se decantează, iar faza solidă se usucă, totodată densitatea optică a extractului se determină la 404 nm şi grosimea stratului de absorbţie de 0,1 cm, iar cantitatea de flavonoizi se exprimă prin cantitatea de quercetină. According to the claimed process, the extraction of the plant raw material with 70…75% ethanol is performed, treatment with aluminum chloride, photometric determination of the optical density of the extract and calculation of the amount of flavonoids, and before extraction with ethanol the raw material is extracted with chloroform, after which the chloroform is decanted and the solid phase is dried, at the same time the optical density of the extract is determined at 404 nm and the thickness of the absorption layer is 0.1 cm, and the amount of flavonoids is expressed by the amount of quercetin.
Avantajele invenţiei propuse constau în aceea că la extracţia preliminară cu cloroform se reuşeşte lichidarea influenţei componentelor secundare, inclusiv a clorofilei, asupra determinării flavonoizilor, aplicarea unui exces mare de etanol 70…75% permite extragerea completă a FL şi simplificarea procedurii de determinare; fotometria extractului cu adaos de AlCl3 în punctul isosbestic la 404 nm, unde se intersectează curbele de absorbţie ale complecşilor de rutină şi quercetină cu AlCl3 permite de a calcula printr-un mod mai comod şi de a exprima cu exactitate rezultatele de determinare a FL într-un extract din materie primă vegetală. The advantages of the proposed invention are that preliminary extraction with chloroform eliminates the influence of secondary components, including chlorophyll, on the determination of flavonoids; the application of a large excess of 70…75% ethanol allows for complete extraction of FL and simplification of the determination procedure; photometry of the extract with the addition of AlCl3 at the isosbestic point at 404 nm, where the absorption curves of the rutin and quercetin complexes with AlCl3 intersect, allows for a more convenient calculation and accurate expression of the results of FL determination in an extract from plant raw material.
Rezultatul invenţiei constă în: simplificarea procedeului de determinare, reducerea numărului de operaţiuni în determinarea FL, micşorarea influenţei compuşilor secundari şi a clorofilei asupra rezultatului analizei. The result of the invention consists in: simplifying the determination procedure, reducing the number of operations in determining FL, reducing the influence of secondary compounds and chlorophyll on the analysis result.
Procedeul de determinare cantitativă a flavonoizilor se realizează în modul următor: la 20 mg de materie primă se toarnă 10…15 ml de cloroform şi se încălzeşte într-un balon conectat la un răcitor cu reflux la temperatura de fierbere timp de 85…95 min. După răcire faza lichidă se decantează. Înainte de eliminarea fazei solide din balon se efectuează uscarea ei într-o etuvă la temperatura de 50…60°C. La faza solidă uscată se toarnă 10 ml de etanol 70…75% şi se efectuează extracţia într-un balon cu răcitor cu reflux la temperatura de fierbere timp de 85…95 min. După răcire, balonul se deconectează de la răcitor. La 3 ml de extract se adaugă 1 ml de soluţie apoasă de AlCl3 0,1 M. The procedure for quantitative determination of flavonoids is carried out as follows: 10…15 ml of chloroform is poured into 20 mg of raw material and heated in a flask connected to a reflux condenser at boiling temperature for 85…95 min. After cooling, the liquid phase is decanted. Before removing the solid phase from the flask, it is dried in an oven at a temperature of 50…60°C. 10 ml of 70…75% ethanol is poured into the dried solid phase and extraction is carried out in a flask with a reflux condenser at boiling temperature for 85…95 min. After cooling, the flask is disconnected from the condenser. To 3 ml of extract, 1 ml of 0.1 M AlCl3 aqueous solution is added.
Peste 20…30 min se fotometrează soluţia complexului flavonoizilor cu AlCl3 la 404 nm la grosimea stratului de absorbţie de 0,1 cm. After 20…30 min, the solution of the flavonoid complex with AlCl3 is photometered at 404 nm at an absorption layer thickness of 0.1 cm.
Rezultatele analizei se exprimă în procente de greutate ale agliconului (quercetină - CQ) prin formula: The analysis results are expressed in weight percentages of the aglycone (quercetin - CQ) by the formula:
, unde: , where:
CQ - cantitatea de quercetină, în %; CQ - the amount of quercetin, in %;
D - densitatea optică a extractului, valori absolute; D - optical density of the extract, absolute values;
n - coeficientul de diluare a extractului alcoolic cu soluţie de clorură de aluminiu, valori absolute; n - dilution coefficient of the alcoholic extract with aluminum chloride solution, absolute values;
V - volumul de etanol, ml; V - volume of ethanol, ml;
m - masa materiei prime, mg; m - mass of raw material, mg;
W - umiditatea materiei prime, %. W - raw material humidity, %.
Umiditatea materiei prime vegetale se determină prin metoda prevăzută în Farmacopeea de Stat [Государственная фармакопея СССР, издание 11-е, том. 1, Медицина, Москва, 1987, с. 285-286]. The moisture content of the plant raw material is determined by the method provided in the State Pharmacopoeia [State Pharmacopoeia of the USSR, 11th edition, vol. 1, Medicine, Moscow, 1987, pp. 285-286].
Recalcularea rezultatului analizei pentru altă flavonoidă rutină se efectuează prin înmulţirea raportului masei moleculare a flavonoidei rutină anhidră C27H30O16 (RUT) la masa moleculară de quercetină anhidră C15H10O7 (CQ), care este aglicon al rutinei: Recalculation of the analysis result for another flavonoid rutin is performed by multiplying the ratio of the molecular mass of the flavonoid anhydrous rutin C27H30O16 (RUT) to the molecular mass of anhydrous quercetin C15H10O7 (CQ), which is the aglycone of rutin:
Masa moleculară a rutinei: Molecular mass of rutin:
MRUT = 610,5 [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280805&loc=ec_rcs]. MRUT = 610.5 [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280805&loc=ec_rcs].
Masa moleculară a quercetinei: Molecular mass of quercetin:
MCQ=302,2 [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280343&loc=ec_rcs]. MCQ=302.2 [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280343&loc=ec_rcs].
La determinarea cantitativă a FL conform procedeului propus la extragerea preliminară a materiei prime vegetale cu cloroform este eliminat cel puţin componentul principal care împiedică la determinarea flavonoizilor - clorofila, care este prezentă în toate plantele, precum şi alţi compuşi secundari. FL din materie primă vegetală nu se extrag cu cloroform. Folosirea unui exces mare de etanol 70…75% (raport 1:500…1000) asigură extragerea completă a FL, exclude procedura ulterioară de diluare a extractului la pregătirea soluţiilor fotometrice, aceasta permite reducerea volumelor extractului (până la 10 ml) şi a soluţiilor fotometrice (până la 4 ml), ce simplifică esenţial procesul de determinare. Extractul cu adaos de AlCl3 se fotometrează în punctul isosbestic la 404 nm, unde se intersectează curbele de absorbţie ale complecşilor de rutină trihidrat 0,001M (RUT·3H2O, M = 664,56) When quantitatively determining FL according to the proposed procedure, preliminary extraction of plant raw material with chloroform eliminates at least the main component that prevents the determination of flavonoids - chlorophyll, which is present in all plants, as well as other secondary compounds. FL from plant raw material is not extracted with chloroform. The use of a large excess of 70…75% ethanol (ratio 1:500…1000) ensures complete extraction of FL, excludes the subsequent procedure of dilution of the extract when preparing photometric solutions, this allows reducing the volumes of the extract (up to 10 ml) and photometric solutions (up to 4 ml), which essentially simplifies the determination process. The extract with the addition of AlCl3 is photometered at the isosbestic point at 404 nm, where the absorption curves of the 0.001M trihydrate rutin complexes (RUT 3H2O, M = 664.56) intersect.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=16218542 http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=16218542
&loc=ec_rcs] şi quercetină dihidrat 0,001M (CQ·2H2O, M = 338,2) [http://pubchem.ncbi.nlm.nih. &loc=ec_rcs] and quercetin dihydrate 0.001M (CQ·2H2O, M = 338.2) [http://pubchem.ncbi.nlm.nih.
gov/summary/summary.cgi?cid=16218542&loc=ec_rcs], obţinuţi după prelucrarea cu soluţie apoasă de 0,1M AlCl3, adică în punctul coeficienţilor molari de extracţie egali ai lor ε404 = 1,716 · 104 1·M-1·cm-1. Se determină concentraţia molară totală a componentelor FL anhidre. Recalcularea conţinutului (%) oricărui component al amestecului de flavonoizi se efectuează prin înmulţirea rezultatului de fotometrie la masa moleculară a flavonoidului-standard. Recalcularea conţinutului de FL în cantitatea sumară de aglicon (quercetină) este cea mai comodă şi precisă, deci în acest caz rezultatele analizei sunt mai puţin dependente de compoziţia FL extractului. De asemenea, activitatea biologică a FL este determinată în principal de aglicon, şi, respectiv, evaluarea farmacologică a extractului se determină de asemenea prin suma totală de aglicon. gov/summary/summary.cgi?cid=16218542&loc=ec_rcs], obtained after processing with 0.1M AlCl3 aqueous solution, i.e. at the point of their equal molar extraction coefficients ε404 = 1.716 · 104 1·M-1·cm-1. The total molar concentration of anhydrous FL components is determined. Recalculation of the content (%) of any component of the flavonoid mixture is carried out by multiplying the photometry result by the molecular mass of the standard flavonoid. Recalculation of the FL content in the total amount of aglycone (quercetin) is the most convenient and accurate, so in this case the analysis results are less dependent on the FL composition of the extract. Also, the biological activity of FL is determined mainly by the aglycone, and, respectively, the pharmacological evaluation of the extract is also determined by the total amount of aglycone.
Exemplu Example
La 20,27 mg de frunze uscate de sunătoare (m), cu conţinutul de umiditate 6% (W) se toarnă 10 ml de cloroform şi se încălzeşte într-un balon conectat la un răcitor cu reflux la temperatura de fierbere timp de 90 min. După răcire faza lichidă se decantează. Faza solidă rămasă într-un balon se spală cu 5 ml de cloroform. Neeliminând faza solidă din balon se efectuează uscarea ei într-o etuvă la temperatura de 50…60°C. La faza solidă uscată se toarnă 10 ml de etanol 70% (V), se încălzeşte într-un balon cu răcitor cu reflux la temperatura de fierbere timp de 90 min. După răcire balonul se deconectează de la răcitor. La 3 ml de extract se adaugă 1 ml de soluţie apoasă de AlCl3 0,1 M. n = 4/3 = 1,33 - coeficientul de diluare a extractului alcoolic după adăugarea la el a soluţiei de AlCl3. Peste 20 min se măsoară densitatea optică a soluţiei complexului flavonoizilor cu AlCl3 la 404 nm la grosimea stratului de absorbţie de 0,1 cm. A fost obţinută valoarea D = 0,207. To 20.27 mg of dried St. John's wort leaves (m), with a moisture content of 6% (W), pour 10 ml of chloroform and heat in a flask connected to a reflux condenser at boiling temperature for 90 min. After cooling, the liquid phase is decanted. The solid phase remaining in a flask is washed with 5 ml of chloroform. Without removing the solid phase from the flask, it is dried in an oven at a temperature of 50…60°C. To the dried solid phase, pour 10 ml of 70% ethanol (V), heat in a flask with a reflux condenser at boiling temperature for 90 min. After cooling, the flask is disconnected from the condenser. To 3 ml of extract add 1 ml of 0.1 M aqueous solution of AlCl3. n = 4/3 = 1.33 - the dilution coefficient of the alcoholic extract after adding the AlCl3 solution to it. After 20 min, the optical density of the solution of the flavonoid complex with AlCl3 is measured at 404 nm at an absorption layer thickness of 0.1 cm. The value D = 0.207 was obtained.
Conţinutul FL în recalcul pe quercetină se calculează după formula sus-menţionată. Conţinutul flavonoizilor în frunzele de sunătoare a constituit 2,43%. După recalcularea pe rutină conţinutul FL a constituit 5,13%. The FL content in recalculation on quercetin is calculated according to the above-mentioned formula. The flavonoid content in St. John's wort leaves was 2.43%. After recalculation on rutin, the FL content was 5.13%.
Conţinutul FL în petalele florilor de sunătoare a constituit 3,19%. După recalcularea pe rutină conţinutul FL a constituit 6,44%. The FL content in the petals of St. John's wort flowers was 3.19%. After routine recalculation, the FL content was 6.44%.
Cercetările au fost efectuate în Centrul Ştiinţific în Domeniul Medicamentului al USMF „Nicolae Testemiţanu” şi în Laboratorul Metode Fizico-Chimice de Cercetare şi Analiză al Institutului de Chimie al AŞM. Au fost efectuate 34 de cercetări experimentale. The research was conducted in the Scientific Center for Medicine of the "Nicolae Testemiţanu" State University of Medicine and Pharmacy and in the Laboratory of Physico-Chemical Methods for Research and Analysis of the Institute of Chemistry of the Academy of Medical Sciences. 34 experimental researches were conducted.
1. SU 1507394 A1 1989.09.15 1. SU 1507394 A1 1989.09.15
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| Publication number | Priority date | Publication date | Assignee | Title |
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| SU1507394A1 (en) * | 1987-08-25 | 1989-09-15 | Всесоюзный научно-исследовательский институт химии и технологии лекарственных средств | Method of quantitative determination of flavonoids in vegetational stock |
| RU2035460C1 (en) * | 1993-06-29 | 1995-05-20 | Борис Алексеевич Фадеев | Method of quercitin preparing |
| RU2085203C1 (en) * | 1994-07-06 | 1997-07-27 | Александр Владиленович Мазулин | Method of quantitative determination of flavanoids in extractive oils from vegetable raw |
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| SU1507394A1 (en) * | 1987-08-25 | 1989-09-15 | Всесоюзный научно-исследовательский институт химии и технологии лекарственных средств | Method of quantitative determination of flavonoids in vegetational stock |
| RU2035460C1 (en) * | 1993-06-29 | 1995-05-20 | Борис Алексеевич Фадеев | Method of quercitin preparing |
| RU2085203C1 (en) * | 1994-07-06 | 1997-07-27 | Александр Владиленович Мазулин | Method of quantitative determination of flavanoids in extractive oils from vegetable raw |
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