Phytochemical and Toxicological Studies of Zygophyllum album L.f.
Amal M.Y. Moustafa,
Ahmed I. Khodair,
Faiza M. Hammouda
Husseiny A. Husseiny
Investigation of the chemical constituents of Zygophyllum album L.f. (Zygophyllaceae family) let to isolate three flavonoids via Kaempferol, Isorhamnetin and Quercetin-3-O-glucoside, one β-carboline alkaloids; Harmine, 16 n-alkanes (C12-C32), β-amyrin, stigmasterol and β-sitosterol and nine fatty acids. The stucctures of these compounds were established by Mass spectrometry (MS), Gas-liquid chromatography (GLC) and spectroscopic techniques, including Ultra-violet (UV), Infra-red (IR) and Nuclear Magnetic Resonance spectroscopic analysis (1H NMR). The oral LD50±standard error and their 95% fiducial limits of the total alcoholic extract were 5.9±0.25 and (5.59-6.23) g kg-1 bw, respectively. While, the intra peritoneal LD50±standard error and their 95% fiducial limits of the extract were 2.60±0.15 and (2.44-2.77) g kg-1 bw, respectively. The total alcoholic extract of the plant could be highly toxic for rats. The extrapolated calculation to human revealed that, this plant could be considered as slightly toxic for man.
Zygophyllum album L. belongs to Zygophyllaceae family, genus Zygophyllum.
Nine species of Zygophyllum are recorded in Egypt (Täckholm,
This plant used in traditional medicine as a remedy for rheumatism, gout, asthma and as a diuretic. Some Bedouins used it as hay or added it to the dry ration. However, it was found to be toxic to the sheep and caused high mortality (Attia and Samar, 2004).
Previous investigation of Zygophyllum album L.f. revealed that the plant
contains Zygophyllin, β-sitosterol-β-D-glucopyranoside, carbohydrates,
tannins, lactones, proteins/amino acids, saponins, triterpene and flavonoid
glycosides (Attia and Samar, 2004; Hani, 1995; Hassanean, 1993; Hassanean, 1993;
Shoaib, 1957; El-Monayeri et al.,
1981; El-Shourbagy and Kishk, 1975).
The aim of this study deals with the chemical study of the plant as regards their constituents particularly alkaloids and flavonoids as well as lipids. The acute toxicity of the total alcoholic extract of the plant was studied to determine the safety margin, qualitative features and quantitative assessment of toxic over dosage. This study was carried out using oral and intra peritoneal administration.
MATERIALS AND METHODS
Fresh aerial parts of Zygophyllum album L.f. (Zygophyllaceae) were
collected from Suez Canal region and Southern Sinai in April during the flowering
stage. The identity was established by Prof. Dr. Moustafa Zaghloul, Prof. of
Floriculture and Medicinal plants, Department of Horticulture, Faculty of Agriculture,
Suez Canal University. A voucher specimen (Number AMYM-1003) has been deposited
in the Herbarium Department of Botany, Faculty of Science, Suez Canal University,
Melting points were determined on Büchi 535 melting point apparatus.
IR spectra were recorded on 1430 Ratio Recording, Perkin-Elmer, IR-data station
Epson FX-86e, in KBr disks. UV spectra were obtained on lambda 4B UV/Vis spectrophotometer
(Perkin-Elmer) in the region of 190-900 nm. EIMS (ionization voltage 70 ev)
was measured on HP-Model, MS-5988. High resolution, Inlet type-DIP, final temperature
200°C and was measured on GC-MSQP 1000EX Schimädzu. 1H NMR
was carried out in d6-dimethylsulphoxide, using Jeol JNM-EX 270 FT
NMR system, operating at AMX 270 MHZ. The chemical shifts are given in ppm (δ),
relative to TMS as internal standard and coupling constants are in Hz. The fractions
obtained were subjected to gas-liquid chromatographic analysis (PYE UNICAM Series
304GC), using coiled glass column (2.8 mx0.4 mm I.D.), packed with Diatomite
C (100-120 mesh) and coated with 1% OV-17, programmed at 10°C min-1
from 70 to 270°C, then isothermally at 270°C for 25 min, injector temperature
and FID detector at 300°C and the nitrogen carrier gas at a flow rate of
30 mL min-1. GLC of the methyl esters of the fatty acids was carried
out by PYE UNICAM Series 304 Gas Chromatograph equipment with FID and SGE injector
split mode, using capillary column (25 mx0.22 mm I.D., 0.2 μm thickness)
packed with vitreous silica coated with FFAP (free fatty acid phase), programmed
at 12°C min-1 from 70 to 190°C, injector temperature at 250°C
and FID detector at 270°C and the flow rate of hydrogen is 41.0 cm sec-1.
Extraction, Isolation and Characterization
The upper parts of the plant (leaves, flowers and stems) were air dried
and ground altogether as a fine powder. The phytochemical screening and the
proximate analysis included ash and moisture contents were performed in accordance
with AOAC (1990).
Air dried and powder aerial parts (1.5 kg) of the plant were extracted with
petroleum ether (40-60°C) to yield 35 g of a dark green oily residue (lipid
fraction). The marc was macerated with ethanol (80%) at room temperature till
exhaustion. The resulting alcoholic extract was concentrated to obtain a crude
residue (750 g). The obtained lipid fraction was treated with hot acetone to
effort 8 g of acetone insoluble fraction and 18 g of acetone soluble fraction
as oily material. Ten grams of the last fraction were saponified to yield 4.5
g of yellowish brown, semi-solid residue of unsaponifiable matter and 3.2 g
of semi-solid residue of fatty acids. Three grams of the unsaponifiable matter
were subjected on a neutral alumina column chromatography with petroleum ether
and increasing the polarity with benzene.
The flavonoids were isolated by treating the alcoholic extract of the plant
with hot water, followed by extraction with chloroform (CHCl3) then
ethyl acetate, to effort 7.5 and 11.5 g, respectively.
Fractionation of the flavonoids was affected by PPC and sephadex LH-20 column
chromatography. UV- absorption spectra of the isolated flavonoidal components
Z-I, Z-II and Z-III were measured in methanol (MeOH) as well as in MeOH after
the addition of shift reagents. Acid hydrolysis of the flavonoidal compound
Z-III. Compound (Z-III) (25 mg) was refluxed with10 mL of 10% sulphuric acid
for 3 h, the concentrated mixture was applied on PC (Attyia
and Ashour, 2002).
(50 mg), Rf; [0.76, 0.08, BAW (4:1:5) and 15%AcOH respectively].
IR bands (KBr) vmax; 3494, 1665, 1605, 1590, 1540, 1464, 1377,
1082 and 1029 cm-1. UV λmax nm (MeOH): 269.8, 319sh,
367.8. EIMS m/z (rel. Int.): 286 (100), 285 (33.56), 258 (5.57), 153(4.97),
152 (0.60), 121 (14.97), 93 (4.59).
(45 mg), yellow crystals, m.p. 302-303°C, Rf; [0.64, 0.05, BAW
(4:1:5) and 15% AcOH, respectively]. IR bands (KBr) vmax; 3402, 1660,
1600, 1587, 1515, 1450, 1091 and 1027 cm-1. UV λmax
nm (MeOH): 276.6, 372. EIMS m/z (rel. Int.): 316 (100), 315 (23.60), 288 (2.70),
153 (5.70), 152 (4.30), 151 (4.40) and 123 (4.40). 1H NMR (270 MHZ,
d6-DMSO): δ 6.15 (1H, d, H-6), 6.45 (1H, d, H-8), 7.65 (1H,
d, H-6`), 7.8 (1H, s, H-2`), 6.9 (1H, d, H-5`), 3.85 (3H, s, OMe at C-3`), 12.5
(1H, s, OH C-3), 9.5 (1H, s, OH at C-7), 9.8 (1H, s, OH at C-4`), 10.8 (1H,
s, OH at C-5) which are exchangeable.
(30 mg), Rf; [0.56, BAW (4:1:5)]. UV λmax nm (MeOH):
255, 258, 300sh, 347, 356. PC analysis of Z-III residue after hydrolysis revealed
the presence of glucose as a sugar moiety.
The study of alkaloids of the investigated plant was carried out after defatting
by percolating with ethanol at room temperature till exhaustion then concentrating
and acidifying with HCl. The acidic solution was extracted with ether and then
reduced with zinc dust. The solution was rendered alkaline with NH4OH.
The alkaloidal fraction was extracted with CHCl3 to yield 1.76 g
of yellowish brown residue which represented 0.12% total crude alkaloids of
the dry plant material. Fractionation of the major alkaloidal constituent Z-IV
was affected by PTLC using solvent CHCl3-MeOH (85:15).
(32 mg), Rf; [0.63, CHCl3-MeOH (4:1:5)]. EIMS m/z (rel.
Int.): 212 (100), 211 (6.13), 197 (29.18), 184 (2.35), 183 (7.43), 182 (2.26),
181(2.59), 155 (0.93), 154 (2.35), 127 (4.37). 1H NMR (270 MHZ, d6-DMSO):
δ. 11.45 (1H, s, -NH), 8.25 (1H, d, H-3), 8.05 (1H, d, H-6), 7.8 (1H, d,
H-4), 7.05 (1H, d, H-8), 6.8 (1H, q, H-5), 3.85 (3H, s, OMe at C-7) and 2.75
(3H, s, Me at C-1).
Acute toxicity studies were performed on 128 mature albino male rats and
nearly of the same age and weight (190-210 g), which were obtained from the
animal house of National Research Center. A pilot test was carried out to determine
the range of doses, which were estimated to cause 0-100% deaths in the final
test. Rats were divided into groups, each composed of 8 animals. The test was
carried out during November under a laboratory temperature of about 19-22°C.
The extract of the plant was suspended in dist. water of tween 80 (polyoxyethylene
sorbitan) (3:1) V/V for 1 g of alcohol extract. After oral and intra peritoneal
administration of the extract, the animals were kept under observation during
the following 24 h. Deaths occurring during this period were recorded in 24,
48 and 72 h for each group. Dose mortality curves were constructed covering
responses between 0-100% lethalities. Toxicity data were analyzed in accordance
with Behrens and Karber (1935) and statistically by
Litchfield and Wilcoxon (1949) procedure. The latter
method also allowed comparison of the oral and intra peritoneal dose mortality
curves with respect to their slope function, parallelism and relative toxicity
RESULTS AND DISCUSSION
The results obtained from the preliminary phytochemical screening of Zygophyllum album L.f. revealed the presence of alkaloids, flavonoids and saponins as major components. Also, the presence of carbohydrates and/or glycosides, coumarins, sterols and/or triterpenes, tannins and cardiac glycosides. The average percentages of the constants of the plant were calculated as shown in Table 1.
The results obtained from the GLC chromatogram of hydrocarbon fraction, eluted
with petroleum ether-benzene (90:10), revealed the presence of a series of 16
n-alkanes (n-C12 to n-C32) with C26H54
as the major constituent of the mixture (19.36%) and the percent of the other
constituents ranging from 0.39-11.13% as shown Table 2 and
Fig. 1. Odd-numbered n-alkanes were not the major constituents
in Zygophyllum album L.f. Therefore, selection of morphologically uniform
samples for chemotaxonomic comparison of n-alkane content is essential for search.
As similar to the hydrocarbon content of Cassia obtusifolia L. which
give even-number n-alkanes as major constituents (Wilkinson,
1970). This phenomenon is unusual comparing with the distribution of n-alkanes
in angiosperm in which, the predominant n-alkanes have odd-carbon numbers (Chikaraishi
and Naraoka, 1982; Ohkouchi et al., 2000;
Disnar and Harouna, 1994).
The residue obtained from fractions eluted with petroleum ether-benzene (75:25)
gave white crystalline needles in MeOH. It melted at 197-199°C both alone
and upon admixture with authentic β-amyrin. It gives positive Liebermann-Burchardt
reaction, indicating that it is a triterpenoid in nature. TLC on Silica gel
using different solvent systems showed to be a single spot possessing the same
Rf as authentic β-amyrin. The IR-spectrum of the isolated substance
showed the same absorption bands characteristic for authentic β-amyrin
(Barreiros et al., 2002).
|| Constants of Zygophyllum album L
|1Average percentages (means of duplicate analysis),
2Calculated on the air-dried plant material, 3Calculated
on the oven-dried plant material
|| Gas liquid chromatographic analysis of the hydrocarbon fraction
|| GLC chromatogram of hydrocarbon fraction
The combined fractions eluted with petroleum ether-benzene (50:50) gave white
crystalline needles in ethanol, m.p. 133-135°C. The substance showed a single
spot on TLC, using toluene- acetone system (90:10), possessing Rf;
0.44. It was identical with authentic β-sitosterol. However, several authors
(Ghaleb et al., 1972; Bennett
and Erich Heftmann, 1966; Marsili and Morelli, 1968;
Singh et al., 1970; Nagasampagi
et al., 1971) reported that the sterols isolated from plants, showed
single substances on TLC and their physical properties confirmed this. Yet on
subjecting to GLC they were proved to be mixtures. So the sterol fraction was
subjected to GLC. The obtained chromatogram revealed that, the isolated sterol
fraction is a mixture of two compounds possessing the same retention times (18.21
and 20.02, respectively) as authentic Stigmasterol and β-sitosterol (Tian-Jye
et al., 1999). The percentages of them were 74.24 and 25.76%, respectively
from the total sterol fraction.
GLC chromatogram of the methyl ester of fatty acids revealed the presence of nine fatty acids according to their retention times in comparison with authentic. The saturated fatty acids represent 92.68% while the unsaturated fatty acids represented by 7.32% of the total fatty acids. The major fatty acids were decanoic acid 33.46% and palmitic acid 31.18% as shown in Table 3 and Fig. 2.
PC of the CHCl3 extract was found to give better separation revealing
the presence of one major flavonoid Z-I Rf; (0.76, BAW 4:1:5), beside
others as minor constituents. The ethyl acetate (EtOAc) extract was found to
contain two flavonoidal spots Z-II, Z-III Rf; (0.64, 0.56, BAW 4:1:5),
beside others as minors. Compound (Z-I), UV-spectrum of Z-I in MeOH, (Table
4 and Fig. 3) exhibited λmax at 367.8
nm (band-I) and band-II at 269.8 nm. This indicates that it is a flavonol type
(Mabry et al., 1970). The NaOMe spectrum showed
a bathochromic shift of 54 nm in band-I with a marked decrease in its intensity
and with rapid degeneration indicating to the presence of 3-OH groups in 3,
3', 4'-positions. Also, showed a shoulder peak at 320 nm, which is an indicative
to the presence of 7-hydroxy group. AlCl3 spectrum showed a bathochromic
shift (54.6 nm) indicating the existence of ortho-dihydroxy system and/or
3, 5-di-hydroxy groups. However, AlCl3/HCl spectrum exhibited slightly
hypsochromic shift (3.9 nm) in band-I relative to AlCl3 spectrum,
which is an indication of the presence of 3, 5-OH groups. NaOAc spectrum showed
a bathochromic shift (5.4 nm) in band-II, MeOH/NaOMe, (b) MeOH, MeOH/AlCl3
and MeOH/AlCl3/HCl and (c) MeOH, MeOH/NaOAc and MeOH/NaOAc/H3BO3
indicates the presence of free 7-OH group, also a bathochromic shift (27.5
nm) in band-I with slow degeneration indicating the presence of 3, 3', 4'-trihydroxy
|| Gas liquid chromatographic analysis of the methyl esters
of fatty acids fraction
|| GLC chromatogram of saponifiable fraction
||UV/Vis absorption spectra of compound Z-I in (a) MeOH and
IR-spectrum of Z-I showed a strong band at 3494 cm-1 corresponding
to -OH group, band at 1665 cm-1 corresponding to an α, β-unsaturated
ketone (C = O group of γ-pyrone), absorption bands at 1605, 1590 and 1540
cm-1 (an aromatic system); in addition to the absorption bands at
1464, 1377, 1082 and 1029 cm-1. MS of Z-I showed a molecular ion
(M+) at m/z 286 (100%), which corresponds to the molecular formula
C15H10O6 of four hydroxy substitution patterns.
Hence, the fragmentation pathway undergoes the Retero-Diels Alder reaction-giving
rise to ring-A fragment at m/z 153 (4.97) and m/z 152 (0.60). However, the hydrogen
transfer ion at m/z 153 is much intense than that of the normal fragment ion
at m/z 152, indicating that it has 5,7-di-hydroxy groups. Furthermore, loss
of CO directly from the molecular ion (M+-CO) was also shown, leading
to the phenylbenzofuran fragment ion at m/z 258 (5.57) which further fragments
giving rise to the benzoyl ion at m/z 121 (14.97) and lose CO directly giving
m/z 93 (4.59). Moreover, the molecular ion (M+) m/z 286 loses hydrogen
to give the molecular ion (M+-1) at m/z 285 (33.56). The previous
data are in agreement with those reported for kaempferol as shown in Fig.
4 (Hamzah and Lajis, 1998; Coelhoa
et al., 2003; Hadizadeh et al., 2003).
Compound (Z-II), UV-spectrum in MeOH, (Table 4 and
Fig. 5) exhibited a λmax at 372 nm (band-I)
and band-II at 276.6 nm which indicates that it is a flavonol type (Mabry
et al., 1970). The results obtained from UV-spectrum, IR-spectrum,
MS and 1H NMR are in agreement with those reported for Isorhamnetin
as shown in Fig. 4 (Shahat et al.,
Compound (Z-III), UV-spectrum in MeOH, as shown in Table
4 and Fig. 6 exhibited a band of max. absorbance at 356
nm (band-I) and a shoulder band at 300 nm indicating that it is a flavonol in
nature. The Rf-values as well as the color under UV-light showed
that, it is probably a flavonoidal glycoside in nature. PC analysis of Z-III
residue after hydrolysis revealed the presence of glucose as a sugar moiety
while the aglycone was identified as quercetin by comparison with authentic
reference. The previous data are in agreement with those reported for quercetin
3-O-glucoside (isoquercetrin) (Assaf, 1980; Aderogba
et al., 2005).
|| Ultra-violet absorption spectra of the isolated flavonoidal
|| Structures of isolated flavonoids
||UV/V is absorption spectra of compound Z-II in (a) MeOH and
MeOH/NaOMe, (b) MeOH, MeOH/AlCl3 and MeOH/AlCl3/HCl
and (c) MeOH, MeOH/NaOAc and MeOH/NaOAc/H3BO3
||UV/V is absorption spectra of compound Z-III in (a) MeOH and
MeOH/NaOMe, (b) MeOH, MeOH/AlCl3 and MeOH/AlCl3/HCl
and (c) MeOH, MeOH /NaOAc and MeOH/NaOAc/H3BO3
The major alkaloid spot Z-IV (Rf; 0.63, CHCl3-MeOH (85:15))
appeared as fluorescent blue under UV-light before spraying the reagent and
colored bright orange red after spaying. The other alkaloidal spots appeared
as minor orange spots on a yellow background with Rf; 0.83, 0.57,
0.47, 0.35, 0.28, 0.25 and 0.17. Compound (Z-IV) (30 mg), the data obtained
from MS and 1H NMR are in agreement with those reported for harmine
(7-methoxy-1-methyl-β-carboline) as shown in Fig. 7 (Olga
et al., 2005; Baisa, 2002).
||Structure of isolated Harmine
The qualitative features recorded during the 24 h period following oral and
i.p. administration with steadily progressive increases in test doses of total
alcoholic extract of the plant resulted in: Behavioral; gradual cessation of
spontaneous motility, ataxia and partial tail elevation, skeletal muscle hypotonia,
loss of rigidity reflex and motor incapacitation, progressive sedation, eventually
passing into hypnosis verified by closure of eyelids, analgesia, lack of response
to painful stimuli and tail compression. Respiration; tachyponea, alternating
with phases of apnea, dyspnea with irregular breathing and terminal gasping
respiration. Heart; feeble cardiac contractions, associated with cyanosis, asphysine,
convulsive fits ending by cardiorespiratory failure. Itching; presumably reflecting
histamine release from H1-receptors in cutaneous mast cells. Diarrhoea
and loose stools excessive urination evidenced by wetting of perineal region.
Well-marked swelling and engorgement of testicles with diffuse hyperaemia and
congestion of the skin of the scrotum. Analgesia arises from antagonistic interactions.
This type of interactions is responsible for the blocking of endogenous hypothalamic
opioid receptors. Likewise, tail elevation (strobes phenomenon) constituting
could be considered as a cardinal sign of agonist/antagonist interaction with
opioid receptors in alimentary tract. According to Zbindin
(1963), tests inhibition of spermatogenesis is enhanced particularly in
the stage of transformation from spermatid to immature spermatozoa. It is apparent
from the obtained toxic manifestations and comparing them with the standard
manifestation of acute toxicity by Häyes (1989),
Zygophyllum album L.f. extract has mast cell H1 histamine
releasing property from skin, autonomic, CNS and neuromuscular effects. These
findings are in agreement with those recorded by Steyn and
Liebig (Steyn, 1929 ; Liebig et al., 1974),
who studied the toxic activity of some species of the genus Zygophyllum.
On the other hand, the LD50 of the alcoholic extract of the plant
was achieved by two methods of analysis for 24 h, to allow a comparison between
them. By using the method of Behrens, LD50 of the plant extract for
oral and i.p administration were 5.975 and 2.575 g kg-1 body weight,
respectively, as shown in Table 5 and 6.
It is remarked that, some rats which did not die in the first 24 h might survive
and escape the fatal outcome, but others might die in the second or third day,
which shown in Table 7 and 8. Whereas, oral
LD50±standard error and their 95% fiducial limits of the extract
calculated by Lichfield procedure were 5.9±0.25 and (5.5924-6.2245) g
kg-1 bw., respectively. While, the intra peritoneal LD50±standard
error and their 95% fiducial limits of the extract were 2.60±0.15 and
(2.4413-2.7690) g kg-1 bw., respectively. Details of calculations
of LD50 and its 95% fiducial limits are shown in Table
9 and 10.
||The effect of oral route administration of different doses
of the alcoholic extract of Zygophyllum album L.f. on the mortality
rate in rats
||The effect of intra peritoneal injection of different doses
of the alcoholic extract of Zygophyllum album L.f. on the mortality
rate in rats
||Oral toxicity of the total alcoholic extract of Zygophyllum
album L.f. in adult normal male albino rats showing the number
of animals that died during 72 h after oral administration of the extract.
Each group consists of 8 rats
||Intra-peritoneal (i.p.) toxicity of the total alcoholic extract
of Zygophyllum album L.f. in adult normal male albino rats
showing the number of animals that died during 72 h after (i.p.) administration
of the extract. Each group consists of 8 rats
||Acute oral toxicity of total alcoholic extract of Zygophyllum
album L.f. in adult normal male albino rats. Body weight range
of rats 190-210 g; average 200 g. Graded test doses of Zygophyllum album
L.f. administered by intra gastric intubation of 100 g % w/v. aqueous solution
(incorporating tween 80 in proportion of 3:1) in volumes not exceeding than
1.6 mL per rat. Animals observed for mortality over 24 h following oral
administration. Data obtained by Lichfield and Wilcoxon (1949) procedure
The oral and intra peritoneal LD50 values of the plant obtained
by Behrens are enclosed within the 95% confidence limits of acute LD50
determined by Lichfielde procedure indicating the practical equality of acute
lethal toxicity of the total alcoholic extract obtained by the two methods.
The obtained results point out that, the dose mortality lines after oral and
i.p. administration are parallel and the toxicity ratio of the oral to i.p.
LD50 is about 2.2692 as shown in Table 11.
In other words, the total extract of the plant is about 2.2692 times toxic
when injected i.p. than in oral administration of such an extract. The oral
and i.p. LD50 of the total alcoholic extract of the plant, in rats
was extrapolated to human and a variety of other mammalian species (e.g., guinea-pigs,
monkeys, ....etc.) (Paget and Barnes, 1964), Table
12. Moreover, the three conventual oral lethal doses and 95% confidence
limits of acute oral LD50 in rats expressed per gram weight of the
crude powdered plant were extrapolated to four species of farm animals (goats,
sheep, cows and buffaloes). The data obtained according to Mellet
(1969) and Krupp (1982), revised were tabulated in
||Acute Intra peritoneal (i.p.) toxicity of total alcoholic
extract of Zygophyllum album L.f. in adult normal male albino
rats. Body weight range of rats 190-210 g; average 200 g. Graded test doses
of Zygophyllum album L.f. administered by intra gastric intubation
of 100 g% w/v. aqueous solution (incorporating tween 80 in proportion of
3:1) in volumes not exceeding than 1.6 mL per rat. Animals observed for
mortality over 24 h following i.p. administration. Data obtained by Lichfield
and Wilcoxon (1949) procedure of calculation
||Compiled data showing test of parallelism of best-fitting
log. dose probit mortality lines of acute oral and intra peritoneal i.p.
lethal toxicities of the total alcoholic extract of Zygophyllum album
L.f. in adult ormal male albino rats and the statistical significance
of lethal toxicities between two routes of systemic administration. Data
obtained by Litchfield and Wilcoxon (1949) procedure
|*From Nomograph No. 4, #Significant
||Hypothetical values for the intra peritoneal and oral median
lethal doses (LD50) and 95% fiducial limits of the total alcoholic
extract of Zygophyllum album L.f. in a variety of mammalian species
as obtained by Paget and Barnes method (1964)
||Extrapolated values for three conventual oral lethal toxic
doses and 95% confidence limits of acute oral LD50 expressed
per gram weight of the crude powdered plant in four species of farm animals
(interspecies dosage conversion, according to revised Mellet,
From the public health point of view, it is important to extrapolate the oral
LD50 in rats to man using the method of Paget and
Barnes (1964). The results registered in Table 12 showed
that, the oral LD50 and their 95% confidence limits in man was found
to be 0.944 (0.8948-0.9959) g kg-1 b.w. The obtained results revealed
that, the total alcoholic extract of the plant could be considered as slightly
toxic to man. About 7.5-8.4 g kg-1 b.w. of crude powdered plant was
found to be toxic to the sheep and caused high mortality. These results are
in agreement with Steyn, (1929, 1933
and 1934) and Watt and Breyer-Brandwijk
(1962) who recorded that, Zygophyllum microcarpum Cham. and Schlect.
was toxic and caused higher mortality in sheep.
Investigation of Zygophyllum album L.f. resulted in the isolation and identification of flavonoids, β-carboline alkaloids and lipid constituents with percentages 1.27, 0.12 and 2.33% relative to the total powder plant. The flavonoid constituents include Kaempferol, Isorhamnetin and Quercetin-3-O-glucoside as the major constituents. Harmine was isolated and identified as the major constituent of β-carboline alkaloids. The isolation and identification of lipid fraction resulted in 16 n-alkanes (C12-C32) with C26H54 as the major constituent and β-amyrin, stigmasterol, β-sitosterol and nine fatty acids. The saturated fatty acids represent 92.68% while the unsaturated fatty acids represented by 7.32% of the total fatty acids. The major fatty acids were decanoic acid 33.46% and palmitic acid 31.18%.
It is apparent from the obtained toxic data and comparing them with the standard manifestation of acute toxicity that the total alcoholic extract of the plant could be highly toxic for rats. The oral LD50±standard error and their 95% fiducial limits of the total alcoholic extract for rats were 5.9±0.25 and 5.59-6.22 g kg-1 b.w., respectively. The intra peritoneal LD50±standard error and their 95% fiducial limits of the total alcoholic extract were 2.60±0.15 and 2.44-2.77 g kg-1 b.w., respectively. The calculated percentage of crude powdered plant was toxic to sheep (7.5-8.4 g kg -1 b.w.) and caused high mortality. The extrapolated calculation to human, revealed that, this plant could be considered as slightly toxic for man.
We thank Prof. Dr. Shams I. Ismail, pharmaceutical sciences department, National research center; for his indispensable help during the whole work and for supplying the facilities during the work and Prof. Dr. H. A. Gammaz, pharmacology department, faculty of veterinary medicine, Suez canal university, for his kind help in executing the toxicological studies.
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