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1984 Afr. J. Pharm. Pharmacol. Table 1. Yields (%) of C. colocynthis Schrad. leaf extracts. Chemical Aque Petroleum Chlorofo Ethyl Acet Metha group ous ether rm acetate one nol Yield (%) 11.047 2.046 8.874 1.342 7.376 13.304 Table 2. Preliminary phytochemical screening of C. colocynthis Schrad. leaf extracts. Chemical group Aque ous Petroleum ether Chlorofo rm Ethyl acetate Acet one Alkaloïds + - - + + + Coumarins + - - + - - Flavonoïds - - - - - - Anthracenic heterosids - - - - - - Cardiotonic heterosids - - - - - - Iridoïds + - - - - + Saponosids - - - - - - Steroïds + + - - - - Gallic tannins + - + + - - plasma, just before adding the thromboplastin-calcium reagent. All experiments were carried out in at least, triplicates. The method described by Brown (1988) suitably modified, was used for the partial thromboplastin time tests. Alexin® (Sigma Diagnostics), which is the partial thromboplastin with activator, and calcium chloride (0.02 M) were prewarmed to 37°C separately in a water bath. 50 μl of plasma was placed in a test tube. After incubating for 180 s in the water bath, 50 μl of Alexin® was added, and the contents were mixed rapidly. The mixture was then incubated for another 180 s, after which 50 μl of serum, then 50 μl of the prewarmed calcium chloride solution was added while simultaneously starting a timer. The tube was then allowed to remain in the water bath while gently tilting the test tube every 5 s. At the end of 20 s, the test tube was removed from the water, wiped clean with dry gauze, and gently tilted back and forth until a clot was seen and the time recorded. For the test samples, 50 μl of the material was added to the contents of the test tube just prior to the addition of calcium chloride, and readings taken as before. For the two tests, serum and heparin (183 IU/mg) were, respectively, used as negative and positive controls. RESULTS Extraction yields and phytohemical screening Results are shown in Tables 1 and 2. Lyophilised aquous extract and methanol extract showed the better yields. The phytochemical screening showed a significant difference between the tested extracts. We note that the present or the absence of all phytochemical groups is related with the extract polarities and the chemical group types. Antimicrobial activity The in vitro antimicrobial activity of C. colocynthis Metha nol extracts against the microorganisms employed and their activity potentials were qualitatively and quantitatively assessed by MIC and MBC/MFC values. According to the results shown in Tables 3 and 4, aqueous extract of C. colocynthis had great potential of antimicrobial activities against all bacteria and Candida species tested. Apolar to polar fractions of the Soxhlet extracts were also found to be effective against all microorganisms examined. MIC and MBC values for bacterial strains, which were sensitive to the aqueous extract and organic ones of C. colocynthis, were in the range of 6.343 to 1625 μg/ml and 12.687 to 3250 μg/ml, respectively. The maximum was obtained with the ethyl acetate extract against all bacteria exept S. aureus. MIC and MFC values of the Candida species sensitive to the tested extracts were < 6.343 to 1625 μg/ml and 6.343 to 3250 μg/ml, respectively. The best results were observed with acetone and methanol extracts against C. albicans and also with ethyl acetate extract against C. Kreusei (CMI < 6.347 μg/ml and CMF = 6.347 μg/ml). Based on these results, Soxhlet extracts have a stronger activity and broader spectrum than the aquous one. As emphasized elsewhere, Gram-negative bacteria are more sensitive than Gram-positive ones (Marzouk et al., 2010a). As well, basing on the cell wall differences of bacteria, results show that Soxhlet extracts did possess the same selective antimicrobial activity. By inhibiting the growth of all human Candida tested, C. colocynthis Schrad. exerted a broad anticandidal spectrum. The antimicrobial nature of this studied plant organ is apparently related to their contents, particularly alkaloids and iridoids, and this finding is in agreement with a
Table 3. Antibacterial MIC (µg/ml) and MBC (µg /ml) of C. colocynthis Schrad. leaves. Strain E. coli ATCC 25922 P. aeruginosa ATCC 27853 S. aureus ATCC 25923 E. faecalis ATCC 29212 Concentrati on Aque ous Petroleum ether Chlorofo rm Ethyl acetate MIC 1625 12.687 12.687 6.343 MBC 3250 25.375 25.375 12.687 MIC 812 12.687 25.375 6.343 MBC 1625 25.375 50.750 12.687 MIC 1625 25.375 25.375 25.375 MBC 3250 50.750 50.750 101.500 MIC 1625 25.375 25.375 6.343 MBC 3250 50.750 50.750 12.687 Marzouk et al. 1985 Acet one 12.68 7 25.37 5 12.68 7 25.37 5 12.68 7 25.37 5 25.37 5 50.75 0 Metha nol 12.687 25.375 12.687 25.375 12.687 25.375 25.375 25.375 MIC positive control, Levofloxacin (E. coli 0.61 µg/ml, P. aeruginosa 0.3 µg/ml, S. aureus 0.3 µg/ml, E. faecalis 1.22 µg/ml); E. coli, Echerichia coli; P. aeruginosa, Pseudomonas aeruginosa; S. aureus, Staphylococcus aureus; E. faecalis, Enterococcus faecalis. Table 4. Antifungal MIC (µg/ml) and MFC (µg/ml) of C. colocynthis Schrad. leaves. Strain C. albicans ATCC 90028 C. glabrata ATCC 90030 C. kreusei ATCC 6258 C. parapsilosis ATCC 22019 Concentrati on Aque ous Petroleum ether Chlorofo rm Ethyl acetate MIC 1625 12.687 12.687 6.347 Acet one
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African Journal of Pharmacy and Pha
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Editors Sharmilah Pamela Seetulsing
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Minimum inhibitory concentration (M
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Kim et al. 1887 Table 4. Antimicrob
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Klevens RM, Morrison M A, Nadle J,
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Figure 1. Chemical structure of ten
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ng/ml by 4.9 ml blank plasma with 5
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Figure 5. Chromatogram of plasma sp
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Figure 7. Chromatogram of a healthy
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Figure 11. Chromatograms of standar
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Shatoor 1903 Figure 1. The effect o
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Heart rate (Beats/min.) 345 330 315
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Table 2. Percent of changes in hear
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heart rate on coronary flow and car
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Urinary tract infections are the mo
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Table 1. Study characteristics of i
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Table 1. contd. Sun GQ 37 007 Ross
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Study or Subgroup Cai SF 2003 Gao H
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Study or Subgroup Cai SF 2003 Gao H
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cure rate and clinical effective ra
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Table 1. The expression of KAI1 and
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Figure 2. Typical chromatograms of
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Excretion studies Figure 4 shows th
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