Identificarea prin cromatografie pe strat subtire a derivatilor flavonici din Hippophaë fructus

Identificarea prin cromatografie pe strat subtire a derivatilor flavonici din Hippophaë fructus

1. Material si metoda

Prepararea solutiei de analizat: 0.5-1 g produs vegetal se extrag cu 10 ml metanol, timp de 10 min., pe baie de apa la 60⁰C. Solutia filtrata se utilizeaza la analiza cromatografica.

Pregatirea cromatoplacii: se foloseste pulbere de silicagel G Merck 0.25 mm grosime (faza stationara), care se agita in apa distilata (1 g la 2 ml) intr-un vas conic, timp de 1 min., dupa care se intinde in strat subtire pe cromatoplacile de sticla (10 × 15) spalate in prealabil, clatite cu apa distilata si uscate cu alcool; dupa intinderea straturilor, placile se usuca la aer 30 min., apoi se activeaza 45-60 min. in etuva, la 110⁰C.

Aplicarea probelor se face cu ajutorul unei micropipete, linia de start fiind trasata la 2 cm de marginea placii; distanta dintre punctele de start trebuie sa fie de cel putin 1.5 cm, iar cea dintre punctele marginale si latura placii de 2 cm; pentru ca diametrul spoturilor sa nu fie prea mare, se folosesc cromatoplacile calde, solutii extractive concentrate si spotarea in banda; dupa aplicarea spoturilor, cromatoplacile se usuca la aer cateva minute.

Developarea: developantul (faza mobila) se prepara prin amestecarea solventilor in cantitatile corespunzatoare; se introduce in tancul cromatografic; inaltimea lichidului trebuie sa depaseasca 1 cm; tancul se inchide etans si se lasa 30 min. pentru saturarea camerei cu vapori de solvent; placile cromatografice se introduc in pozitie verticala;
se lasa la developat pana cand frontul solventului migreaza 15-18 cm; se delimiteaza linia de start si frontul solventului.

Revelarea: placile examinate initial in UV sunt pulverizate omogen cu solutia indicata (NH₃ vapori), mentinute in etuva 5-10 min., la 110°C, apoi examinate in VIS.

Faza stationara: silicagel G Merck

Faza mobila: toluen-acetat de etil-acid formic (5 : 4 : 1); parcurs util 10 cm.

Revelator ▪ in UV, la 365 nm - flavonele cu hidroxilul de la C₃ liber prezinta o fluorescenta galben-aurie, pana la galben-verzuie; cand hidroxilul de la C₃ lipseste sau este glicozidat, fluorescenta este galben-bruna.

▪ vapori de amoniac - flavonele prezinta culoare (sau fluorescenta) galben-verzuie sau galben-aurie, mai intensa dupa alcalinizare.

Etaloane: solutii metanolice 0.1% de cvercetol si rutozida; se aduc cate 5 μl la punctul de start.

2. Rezultate si discutii

Rezultatele obtinute in urma analizei placilor cromatografice si interpretate pe baza consultarii lucrarilor de specialitate sunt prezentate in Tabelul 4 si Figurile 14-15.

Proba de analizat se caracterizeaza prin zone ale caror coloratii (sau fluorescente) se intensifica dupa tratare cu vapori de amoniac: ▪ galben-verzuie (cvercetol - R_f 0.50) si ▪ galben-aurie (rutozida - R_f 0.53).

Tabelul 4 - Identificarea prin CSS a flavonelor din Hippophaë fructus

  

Figura 14             Figura 15

[1] - Hippophaë fructus (sol. metanolica hidrolizata); [2] - Solutii etalon (0.1% cvercetol, 0.1% rutozida)

Concluzii

1.  Genul Hippophaë cuprinde o singura specie - rhamnoides, care face parte din familia Elaeagnaceae. Catina se prezinta ca un arbust inalt de 1.5-3.5 m, cu numerosi tepi puternici. In functie de conditiile de clima si sol, ea creste diferit si anume ca tufa joasa taratoare, in zonele aride si pe soluri sarace sau sub forma arborescenta, de 8-10 m inaltime, pe solurile fertile. Pe radacinile catinei se formeaza nodozitati fixatoare de azot, ca la leguminoase.

2.  Fructele sunt drupe false, mici, de forma variabila, de la ovoida la globuloasa, chiar turtita. Culoarea lor predominanta este portocalie, cu treceri spre galben. Mai rar apar si fructe de culoare rosie. Pulpa fructelor este de culoare galbena sau portocalie, foarte suculenta si lasa pete unsuroase.

3.  Fructele de catina au un miros placut si aromat, dar nu se consuma cu placere in stare proaspata, fiind acre si astringente. La maturitate completa, ele pierd mult din aciditate si au o aroma particulara, mai puternica atunci cand recoltatul se face dupa inghet.

4.  De la catina se utilizeaza, in scop terapeutic produsul Hippophaë fructus, care contine: vitamine lipo- si hidrosolubile, carotenoide, flavonoide, lipide, acizi triterpenici pentaciclici, fitohormoni, oze, proteine, acizi polifenol-carboxilici (cafeic, clorogenic, chinic), acizi organici, saruri minerale etc.

5.  In fructele de catina se gaseste provitamina A (β-carotenul), care face parte din grupul carotenoidelor, alaturi de α-caroten, fizoxantina, criptoxantina, licopina, zeaxantina. Unul din componentele fructelor de catina alba este uleiul (grasimea bruta), cu un continut ridicat de carotenoide, predominand cele cu valoare de provitamina A, alaturi de substante cu efect regenerator, imunoinductor in vivo.

6.  Fructele de catina au o mare valoare terapeutica, astfel: sunt eficiente in tratamentul bolii reumatismale (complexul de vitamine B); prezinta actiune diuretica, favorizand eliminarea calculilor, mai ales in guta; sunt reconstituante la convalescenti, mai ales dupa viroze; se utilizeaza in tuberculoza si rahitism, in insuficienta renala,
in xeroftalmie si conjunctivite; regleaza circulatia sanguina; mentin si imbunatatesc vederea; sunt antialergice; mentinerea integritatea sistemului nervos central si a scheletului; se utilizeaza in tratamentul hepatitelor si cirozelor hepatice.

7.  De asemenea, catina este utilizata pentru ingrijirea pielii. Datorita actiunii sale revitalizante se foloseste pentru tratarea ranilor, hidratarea tenului, pentru diminuarea durerilor si in tratamentul psoriazisului, arsurilor si ulceratiilor. Siropul si ceaiul de catina au efecte energizante si maresc rezistenta organismului.

8.  Fructele de catina reprezinta o sursa extrem de bogata in vitamine (A, B_1-9, C, E, F, D, P, K), celuloza, proteine, β-caroten, microelemente (P, Ca, Mg, K, Fe, Na), uleiuri complexe etc. In afara de fructe, de la catina se folosesc semintele, frunzele si scoarta, care contin nenumarate substante utile. Uleiul de catina este folosit in tratarea unor afectiuni precum: ulcerul gastric si duodenal, alergiile, diareea, urticaria, reumatismul, afectiunile neuroendocrinologice, circulatorii, hepatice.

9.  De asemenea, se mai foloseste in tratamentul alcoolismului, anemiilor, asteniei si stresului. Se utilizeaza in geriatrie, cu rezultate spectaculoase. Cu extracte de catina se mai trateaza afectiunile oftalmologice, coronariene, hipertensiunea arteriala si gingivitele. In urma unor separari sau concentrari de substante extrase din catina se obtin tratamente extraordinare pentru depresii, boala Parkinson, tumori, adenoame si leucemie. Mugurii de catina au efect afrodisiac. Catina este si un foarte bun antiinflamator si inhiba pofta de mancare in cazul unor tratamente ale obezitatii.

10.   Produsele medicinale obtinute din catina (ceai de fructe, muguri, frunze sau scoarta, sirop sau ulei) sunt complemente alimentare care ajuta organismul sa-si regleze functionarea si sa aiba o imunitate ridicata. Obiceiul de a bea un ceai de catina pe zi poate insemna o sanatate de fier.

11.   De asemenea, siropul poate fi folosit in locul celor din comert care, asa contin nenumarati aditivi alimentari cu efecte negative asupra organismului. Uleiul de catina este insa cel mai folositor datorita faptului ca asigura necesarul de vitamine si oligoelemente.

12.   Partea experimentala a lucrarii cuprinde: studiul histo-anatomic al organelor vegetative la Hippophaë rhamnoides, analiza fitochimica preliminara a produsului Hippophaë fructus si identificarea prin cromatografie pe strat subtire a flavonelor din produsul medicinal Hippophaë fructus

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