Documente Academic
Documente Profesional
Documente Cultură
COALA DOCTORAL
Conductor de doctorat,
Prof. Univ. Dr. STELIAN RADU
Student doctorand,
LIVIU CHIRIGIU
Craiova
2014
Liviu Chirigiu
CUPRINS
CUPRINS
ABREVIERI
INTRODUCERE
STADIUL ACTUAL AL CUNOATERII
CAPITOLUL 1. CONSIDERAII GENERALE PRIVIND SPECIA Sambucus ebulus L.
1.1. ncadrare sistematic
1.2. Distribuia speciei Sambucus ebulus L.
1.3. Chemosistematic
1.4. Potenialul fitoterapeutic
1.4.1. Activitatea antiinflamatoare
1.4.2. Activitatea antioxidativ
1.4.3. Activitatea anti-Helicobacter pylori
1.4.4. Activitatea citotoxic i antiangiogenic
CAPITOLUL 2. METODE DE EXTRACIE I DE ANALIZ A COMPUILOR CHIMICI CONINUI DE
SPECIA Sambucus ebulus L.
2.1. Metode de extracie
2.1.1. Metoda Soxhlet
2.1.2. Extracia cu microunde
2.1.3. Extracia asistat cu ultrasunete
2.2. Metode de identificare i separare a metaboliilor secundari
2.2.1.Gaz-cromatografia cuplat cu spectrometria de mas
2.2.2. Cromatografia de lichide de nalt performan cuplat
2.2.3. Spectrometria de absorbie atomic
CONTRIBUII ORIGINALE
CAPITOLUL 3. IDENTIFICAREA I CARACTERIZAREA PRELIMINAR A SPECIEI
3.1. Caracteristici macroscopice
3.2. Caracteristici microscopice
3.3. Caracteristici microscopice ale pulberilor provenind de la produsele vegetale
3.4. Determinarea umiditii, substanei uscate i reziduului la calcinare pentru specia Sambucus ebulus
3.5. Extracia Soxhlet
3.6. Analiza prin spectroscopia IR cu transformat Fourier (FT-IR)
3.6.1. Materiale i metode folosite
3.6.2. Rezultate i discuii
3.7. Analiza cationilor metalici coninui de specia Sambucus ebulus L. prin SAA
3.7.1. Mineralizarea probelor
3.7.2. Materiale i metode
3.7.3. Rezultate i discuii
CAPITOLUL 4. INVESTIGAII FITOCHIMICE ASUPRA SPECIEI Sambucus ebulus L. PRIN GC-MS I
HPLC / HPLC -MS
4.1. Analiza prin gaz-cromatografie cuplat cu spectrometria de mas a compuilor coninui n
extractele de Sambucus ebulus L.
4.1.1. Pregtirea probelor
4.1.2. Aparatur i condiii de lucru
4.1.3. Rezultate i discuii
4.2. Analiza unor clase de metabolii secundari din Sambucus ebulus prin HPLC i HPLC-MS
4.2.1. Analiza brassicasterolului, campesterolului i stigmasterolului
4.2.1.1. Pregtirea probelor
4.2.2. Separarea antocianilor
4.2.3. Separarea i -amirinului
CAPITOLUL 5. EVALUAREA ACTIVITII BIOLOGICE A UNOR EXTRACTE DIN Sambucus ebulus L.
5.1. Evaluarea activitii antioxidante
5.1.1. Pregtirea probelor
5.1.2. Materiale i metode
5.1.2.1.Coninutul total de compui fenolici
5.1.2.2. Coninutul total de flavonoide
5.1.2.3. Determinarea permanganometric a coninutului n substane organice oxidabile din specia
Sambucus ebulus L.
5.1.2.4. Studii de voltametrie ciclic
5.1.2.5. Determinarea capacitii chelatante a extractelor
5.1.3. Rezultate i discuii
Liviu Chirigiu
1
2
Liviu Chirigiu
CONTRIBUII ORIGINALE
CAPITOLUL 3. IDENTIFICAREA I CARACTERIZAREA
PRELIMINAR A SPECIEI
3.1. Caracteristici macroscopice
n aceast prim etap a cercetrii au fost descrise caracteristicile morfologice
i organoleptice ale organelor vegetale, s-a apreciat aspectul (interior, exterior) i
particularitile caracteristice, dimensiunile, culoarea, mirosul.
Proba de
Sambucus
ebulus L.
Rdcini
Tulpini
Flori
Fructe
Frunze
Masa
iniial
(g)
1,4216
2,1261
1,4758
2,9378
3.3317
Masa dup
uscare (g)
Umiditate
(%)
Reziduu
calcinare
1,2987
1,9463
1,3555
2,5349
3,0672
8,645
8,456
8.8749
13,7143
8.6235
0,0989
0,0860
0,1240
0,0931
0,2536
Cenu
brut
(%)
6,956
4,044
8,402
3,169
7,612
Peres VF, Saffi J, Melecchi MI, Abad FC, Assis Jacques R, Martinez M, Conceic E Caramao EB(2006).
Comparison of soxhlet, ultrasound-assisted and pressurized liquid extraction of terpenes, fatty acids and Vitamin
E from Piper gaudichaudianum Kunth, Journal of Chromatography A, 1105:115118.
Liviu Chirigiu
Principii active
Rdcin
carotenoide
+
uleiuri volatile
+
steroli
+
acizi grai
+
agliconi flavonici
+
alcaloizi baze
+
emodoli
+
- principii active absente.
Frunz
+
+
+
+
+
+
+
Fruct
+
+
+
+
+
+
+
Tulpin
+
+
-
Frunz
+
+
+
-
Fruct
+
+
+
+
-
Tulpin
-
3.435
3.385
3.008
3.008
2.924
2.925
2.926
2.929
2.926
2.853
1.733
1.462
1.378
1.260
1.172
2.854
1.735
1.462
1.377
1.247
1.171
2.854
1.745
1.460
1.377
1.241
1.166
2.854
1.735
1.247
1.171
2.857
1.733
1.457
1.376
1.249
1.170
1.093
1.094
1.098
1.094
1.072
1.023
801
-
802
1.032
723
1.045
801
552
Atribuire
Vibraii ntindere O-H i N-H
(aminoacizi, amine, amide)
C-H(asim) (lipide, acizi grai)4
Vibraii asimetrice de alungire CH3, -CH2(acizi carboxilici)
(sim) grupe CH3
Vibraii ntindere -C=O (esteri)
C-H(ndoire) (a gruprii -CH2 proteine)
Vibraii de deformare (C-H)
C-O, O-H def (polifenoli)
C-O- C, inel
C-H(deformare), C-O,
C-C(ntindere) carbohidrai
C-O (ntindere) polizaharide
C-H (ndoire n afara planului)
Vibraii gruparea fosfat
Schulz H., Baranska M. Identification and quantification of valuable plant substances by IR and Raman spectroscopy,
Vibrational Spectroscopy, 43: 13-25, 2007.
Liviu Chirigiu
Ca
(g/g)
3196,9
2921,8
9539,6
2805,1
Fe
(g/g)
97,1
9,9
148
200,25
Mn
(g/g)
16,4
5,5
49,93
37,1
Mg
(g/g)
1698,6
2127,14
5469
1841,22
Zn
(g/g)
20,7
6,2
23,12
19,46
Pb
(g/g)
1,4
0,78
0,76
0,7
Cr
(g/g)
0,35
0,15
1,37
4,15
Ni
(g/g)
2,5
1,32
0,99
7,146
Cu (g/g)
6,895
4,962
1,923
6,310
7,374
Pb (g/g)
1,021
0,728
0,876
1,952
2,980
Zn (g/g)
33,1
27,9
47,5
26,5
35,6
Produs
Fructe
Frunze
Tulpini
Cd
*
0.32
*
Concentraia (g/g)
Cu
Cr
Zn
2.34
9.85 45.92
5.16
10.27 36.58
2.16
*
16.58
Pb
9.21
12.63
*
Fe
113.40
99.29
*
Chirigiu L., Chirigiu R. G., Tircomnicu V., Bubulica M. V (2011). GC-MS analysis of chemical composition of
Sambucus Ebulus leaves, Chemistry of natural compounds, 47 (1): 126 127, UDC 547.913
Liviu Chirigiu
Formul moecular
C28H46O
C28H48O
C29H48O
Ion [M-H2O+H] +
470
383
395
Fitosterol
tR (min)
Inaltime (mAu)
Brassicasterol
Campesterol
Brassicasterol
Stigmasterol
Campesterol
Campesterol
Brassicasterol
Campesterol
3,59
4,18
3,63
4,46
4,21
4,20
3,31
4,19
174,09
965,29
73,98
163,05
755,37
674,08
221,80
568,67
886,88
2691,67
394,13
952,64
2787,99
2771,32
478,27
2699,82
Concentraia
(mg/100 g)
29,222
136,310
12,418
132,748
109,411
95,118
37,230
80,244
Gabay O, Sanchez C, Salvat C, Chevy F, Breton M, Nourissat G, Wolf C, Jacques C, Berenbaum F (2010).
Osteoarthritis Cartilage, 18:106.
7
Bubulic M.V., Chirigiu L., Popescu M., Simionescu A., Anoaica G., Popescu A (2012). Analysis of sterol
compounds from Sambucus ebulus, Chemistry of natural compounds. 48(3):520-521.
Liviu Chirigiu
S. ebulus frunze
S. ebulus tulpini
S. ebulus rdcini
S. ebulus fructe
Antociani
Delfinidin + pentoze
Petunidin + pentoze
Cianidin 3,5-diglucozide
Delfinidin + pentoze
Petunidin + pentoze
Cianidin 3,5-diglucozide
Petunidin + pentoze
Petunidin + pentoze
Delfinidin + pentoze
Petunidin + pentoze
tR (min)
5,89
6,67
7,22
5,89
6,76
7,29
6,69
6,61
5,72
6,67
Inaltime (mAu)
609,99
2596,46
1025,88
434,31
2387,9
2354,67
2578,0
2832,31
230,88
2688,87
Compus
- amirin
- amirin
- amirin
- amirin
- amirin
- amirin
- amirin
- amirin
tR (min)
3,25
4,23
3,33
4,4
4,00
4,07
3,42
4,07
Inaltime (mAu)
1409,07
229,99
330,73
956,08
2827,83
2840,44
248,86
2786,11
Borneo R, Leon AE, Aguire A, Ribotta P, Cantero JJ (2009). Antioxidant capacity of medicinal plants from the
Province of Cordoba (Argentina) and their in vitro testing in a model food system. Food Chemisty, 112: 664-670.
Liviu Chirigiu
Proba
mprob (g)
VH2SO4 (mL)
VH2O (mL)
10,00
10,00
20,00
20,00
60,00
60,00
118,5
89,5
Nayan R, Rajibul AL, Ismail S, Deepa K, Tirthankar G, Naznin AB (2011). A detailed study on the antioxidant
activity of the stem bark of Dalbergia
sissoo Roxb., an Indian medicinal plant. Food Chem. 126: 1115-1121.
10
Liviu Chirigiu
Tip de extract
Sambucus ebulus
frunze (Se1)
(extract apos)
Sambucus ebulus
fructe (Se2)
(extract apos)
Sambucus ebulus
rdcini (Se3)
(extract apos)
Sambucus ebulus
tulpini (Se4)
(extract apos)
Martor (M1)
Concentraia
2.0 %
1.0 %
0.5 %
0.25 %
2.0 %
1.0 %
0.5 %
0.25 %
2.0 %
1.0 %
0.5 %
0.25 %
2.0 %
1.0 %
0.5 %
0.25 %
-
Ziua 1
0.75
0.80
0.74
1.01
1.00
0.98
1.05
0.82
0.74
0.73
1.21
1.24
0.77
1.00
1.10
0.97
1.23
11
Liviu Chirigiu
K2O
(%)
1,59
1,60
1,63
Substana uscat
(%)
28,50
28,53
28,51
12
Liviu Chirigiu
BIBLIOGRAFIE SELECTIV
1. Iaroenko P D (1962). Geobotanica, Ed. Academiei R.P.R., Bucureti
2. Vernon H (1987). Flowering plants of the World. Andromeda Oxford LTD, Heywood
3. Schulz H., Baranska M. Identification and quantification of valuable plant substances by IR
and Raman spectroscopy, Vibrational Spectroscopy, 43: 13-25, 2007
4. Gabay O, Sanchez C, Salvat C, Chevy F, Breton M, Nourissat G, Wolf C, Jacques C,
Berenbaum F (2010). Osteoarthritis Cartilage, 18:106.
5. Bubulic M.V., Chirigiu L., Popescu M., Simionescu A., Anoaica G., Popescu A (2012).
Analysis of sterol compounds from Sambucus ebulus, Chemistry of natural compounds.
48(3):520-521.
6. Popa T., Bubulic M.V., Chirigiu L., Mogosanu G. D., Popescu R., Popescu H., Researches
upon the heavy metals content of Sambucus nigra L. (Caprifoliaceae), Current Health Sciences
Journal, AprilJune, 2010, 36(2):111114, ISSN 20670656.
7. Chirigiu L, Popescu R., Bubulic M.V., Popescu A (2012). Determination of chromium,
cooper, iron, zinc, cadmium and lead by graphite furnace atomic absorption spectrometry in
seven phytopharmaceutical products, Revista de Chimie,: 63(9): 874-876.
8. Chirigiu L., Chirigiu R. G., Tircomnicu V., Bubulica M. V (2011). GC-MS analysis of
chemical composition of Sambucus Ebulus leaves, Chemistry of natural compounds, 47 (1): 126
127, UDC 547.913
9. Chirigiu L, Bubulica M. V., Chirigiu R. G (2010). GC-MS Analysis of Chemical Compounds
from Stems of Sambucus Ebulus L. Acta Medica Marisiensis, 56(6): 522-525.
70. Patel V.R., Patel P.R., Kajal S.S. Antioxidant Activity of Some Selected Medicinal Plants in
Western Region of India, Advances in Biological Research, 4: 23-26, 2010.
10. Bubulica MV, Chirigiu L., Grumezescu A.M., Popescu A., Simionescu A (2012).
Screening of antioxidant potential of Lonicera tatarica, Viburnum opulus and Sambucus ebulus
L. by multiple in vitro assays. Journal of Medicinal Plants Research Vol. 6(3): 544-552
11. Rop O., Reznicek V., Vasilkova M., Jurikova T., Milcek J., Kramarova D. Antioxidant
properties of European cranberrybush fruit (Viburnum opulus var. edule). Molecules, 15: 44674477, 2010
12. Chirigiu L, Bubulic MV, Averis LME (2012). Assessment of the toxicity of some
phytopharmaceutical products from Caprifoliaceae family by Triticum bioassay, Analele
Universitii din Craiova Seria- Biologie, Horticultur, Tehnologia Prelucrrii Produselor
Agricole, Ingineria Mediului, XVII (LIII): 583-587
13