Using Gas Chromatography-Mass Spectrometry (GC-MS) Technique for Analysis of Bioactive Compounds of Methanolic Leaves extract of Lepidium sativum

 

Hussein J. Hussein1, Imad Hadi Hameed*2, Mohammed Yahya Hadi 3

1Department of Biology, College of Science for women, University of Babylon, Iraq

2College of Nursing, University of Babylon, Iraq

3College of Biotechnology, Al-Qasim Green University, Iraq

*Corresponding Author E-mail: imad_dna@yahoo.com

 

ABSTRACT:

Cress (Lepidium sativum), sometimes referred to as garden cress to distinguish it from similar plants also referred to as cress (from old Germanic cresso which means sharp, spicy), is a rather fast-growing, edible herb. The objective of this study was analysis of the secondary metabolite products. Bioactives are chemical compounds often referred to as secondary metabolites. Ninteeth bioactive compounds were identified in the methanolic extract of Lepidium sativum. The identification of bioactive chemical compounds is based on the peak area, retention time molecular weight and molecular formula. GC-MS analysis of Lepidium sativum revealed the existence of the Glycerin, Monoethanolamine, 1-Deoxy-d-mannitol, 1-Nitro-2-propanol , 2-Butanamine , (S)-,  Furfural, Allyl isothiocyanate , Paromomycin, 2-Hydroxy-2-(5-methylfuran-2-yl)1-phenylethanone , 3,6-Diazahomoadamantan-9-one Hydrazone , 2,3,4-Trimethoxycinnamic acid , 2-Naphthalenol , 2,3,4,4a,5,6,7-octahydro-1,4a-dimethyl-7-(2)- , cis-Vaccenic acid , 9-Octadecenamide , γ-Tocopherol , Phthalic acid , decyl oct-3-yl ester , Ergosta-5,22-dien-3-ol,acetate , ( 3β,22E)- , Campesterol and Cholest-5-en-3-ol ,24-propylidene-,(3β). GC-MS is widely used in pharmaceutical industries for analytical research and development, quality control, quality assurance, production, pilot plants departments for active pharmaceutical ingredients (API), bulk drugs and formulations.

 

KEYWORDS: GC-MS, Bioactive Compounds, Leaves, Lepidium sativum.

 

 


INTRODUCTION:

Lepidium sativum have been widely used to treat a number of diseases in traditional system of medicine throughout Iraq. It is useful in case of lumbago or any other pains about the loins through rheumatism 1-8. Its external application with lime juice for relief of internal inflammation and rheumatic pain is useful. Garden cress is genetically related to watercress and mustard, sharing their peppery, tangy flavor and aroma. In some regions, garden cress is known as mustard and cress, garden pepper cress, pepperwort, pepper grass, or poor man's pepper.

This annual plant can reach a height of 60 cm (~24 inches), with many branches on the upper part. The white to pinkish flowers are only 2 mm (1/12 of an inch) across, clustered in branched racemes. It is useful in asthma, cough, diarrhoea, dysentry, skin disease, blood disorder 9-20. It is having hot and bitter property. It is consider as tonic, galactagogue, aphrodisiac, destroys vata and kapha, cures dysentery, good for pain in abdomen, blood and skin disease and tumours. The fresh fruit is good for injury, skin and eye disease. Its seeds are hot, and leaves hot and dry, diuretic, aphrodisiac, good in inflammation and spleen disease, in chest complaints, bronchitis, and rheumatism and muscular pains. The root is used in secondary syphilis and tenesmus. According to Bellow the seeds are also considered to be galactagogue in the Punjab, and are administered after being boiled with milk, to cause abortion. It is used in the treatment of asthma, cough and bleeding piles, leaves are mildly stimulant and diuretic and useful in scorbutic diseases and liver complaints 21-38. The root is used in secondary syphilis and tenesmus. The seeds of the plant are rubefacient, galactogogue, emmenagogue, laxative, tonic, aphrodisiac and diuretic. They are used in poultice for hurts and sprains. The aim of this study was analysis of the secondary metabolite products of Lepidium sativum.

 

MATERIALS AND METHODS:

Gas chromatography – Mass Spectrum analysis

Interpretation of mass spectrum was conducted using the database of National Institute of Standards and Technology (NIST, USA). The database consists of more than 62,000 patterns of known compounds. The spectrum of the extract was matched with the spectrum of the known components stored in the NIST library. Lepidium sativum GC–MS analysis were carried out in a GC system (Agilent 7890A series, USA). The flow rate of the carrier gas, helium (He) was set to beat 1 mL min−1, split ratio was 1:50. The injector temperature was adjusted at 250◦C, while the detector temperature was fixed to280◦C. The column temperature was kept at 40◦C for 1 min followed by linear programming to raise the temperature from 40◦to 120◦C (at 4 Cº min−1with 2 min hold time), 120 Cº to 170 Cº (at 6 Cº min−1with 1 min hold time) and 170 Cº to 200 Cº (at10◦C min−1with 1 min hold time) 39-52. The transfer line was heated at 280 Cº. Two microliter of FAME sample was injected for analysis. Mass spectra were acquired in scan mode (70 eV); in the range of 50–550 m/z.

 

Statistical analysis

Results of the study were based on analysis of variance (ANOVA) using Statistica Software. A significance level of 0.05 was used for all statistical tests.

 

 


 

Table 1. Major phytochemical compounds identified in methanolic extract of Lepidium sativum.

Serial

No.

Phytochemical compound

RT (min)

Molecular Weight

Exact Mass

Chemical structure

MS Fragment- ions

Pharmacological actions

1

Glycerin

3.31

92

92.047344

 

61,74

anti-inflammatory

2

Monoethanolamine

3.356

61

61.052764

 

61

Anti-Bacterial Agents

3

1-Deoxy-d-mannitol

3.453

166

166.084124

 

61,73,103,166

antispasmodic

4

1-Nitro-2-propanol

3.505

105

105.042593

 

61,69,90,104

antimicrobial

5

2-Butanamine , (S)-

3.59

73

73.0891495

 

58,72

New chemical compound

6

Furfural

3.676

96

96.021129

 

51,67,96

Anti inflammatory activity and analgesic activity

7

Allyl isothiocyanate

3.894

99

99.0142703

 

58,72,84,99

antimicrobial activity

8

Paromomycin

4.414

615

615.296303

 

57,67,80,94,109,191,227,259,292,324

Antimicrobial activity

9

2-Hydroxy-2-(5-methylfuran-2-yl)1-phenylethanone

4.334

216

216.078644

 

51,77,105,122,137,180,214

anticancer activity

10

3,6-Diazahomoadamantan-9-one Hydrazone

8.797

180

180.137497

 

58,72,95,121,180

Anti-microbial

11

2,3,4-Trimethoxycinnamic acid

13.964

238

238.084124

 

53,77,107,163,179,207,238

Anti-stress effects

12

2-Naphthalenol , 2,3,4,4a,5,6,7-octahydro-1,4a-dimethyl-7-(2)-

14.308

238

238.19328

 

55,67,81,107,123,161,177,205,220,238

anti- inflammatory

13

cis-Vaccenic acid

15.423

282

282.25588

 

55,69,83,97,111,123,165,193,222,264,282

anti-asthmatic, anti-inflammatory

14

9-Octadecenamide

17.346

281

281.271864

 

55,72,83,122,220,281

anti- inflammatory action

15

γ-Tocopherol

22.295

416

416.36543

 

57,69,107,151,191,246,288,330,372,416

antioxidant activity

16

Phthalic acid , decyl oct-3-yl ester

21.906

418

418.30831

 

57,104,149,167,193,251,307

anti-bacterial activity

17

Ergosta-5,22-dien-3-ol,acetate , ( 3β,22E)-

23.119

440

440.36543

 

55,67,91,145,213,255,281,327,380

New chemical compound

18

Campesterol

23.543

400

400.370516

 

55,81,145,213,255,289,315,400

anti-inflammatory, and in vitro cytotoxic activities

19

Cholest-5-en-3-ol ,24-propylidene-,(3β)-

24.195

426

426.386166

 

55,69,95,229,281,314,341,393,426

Antibacterial activity

 

 

 

 

Figure 1: GC-MS chromatogram of methanolic extract of Lepidium sativum.

 


 

Figure 2: Mass spectrum of  Glycerin   with Retention Time (RT)= 3.310

 

Figure 3: Mass spectrum of   Monoethanolamine  with Retention Time (RT)= 3.356

 

Figure 4: Mass spectrum of   1-Deoxy-d-mannitol  with Retention Time (RT)= 3.453

 

Figure 5: Mass spectrum of  1-Nitro-2-propanol   with Retention Time (RT)=3.505

 

Figure 6: Mass spectrum of  2-Butanamine , (S)-   with Retention Time (RT)= 3.590

 

 

Figure 7: Mass spectrum of  Furfural   with Retention Time (RT)= 3.676

 

 

Figure 8: Mass spectrum of  Allyl isothiocyanate   with Retention Time (RT)= 3.894

 

Figure 9: Mass spectrum of   Paromomycin  with Retention Time (RT)= 4.414

 

 

Figure 10: Mass spectrum of  2-Hydroxy-2-(5-methylfuran-2-yl)1-phenylethanone   with Retention Time (RT)= 4.334

 

 

Figure 11: Mass spectrum of  3,6-Diazahomoadamantan-9-one Hydrazone with Retention Time (RT)= 8.797

 

Figure 12: Mass spectrum of  2,3,4-Trimethoxycinnamic acid  with Retention Time (RT)= 13.964

 

Figure 13: Mass spectrum of  2-Naphthalenol , 2,3,4,4a,5,6,7-octahydro-1,4a-dimethyl-7-(2)-  with Retention Time (RT)= 14.308

 

Figure 14: Mass spectrum of cis-Vaccenic acid  with Retention Time (RT)=15.423

 

Figure 15: Mass spectrum of  9-Octadecenamide  with Retention Time (RT)=17.346

 

 

Figure 16: Mass spectrum of  γ-Tocopherol  with Retention Time (RT)= 22.295

 

Figure 17: Mass spectrum of  Phthalic acid , decyl oct-3-yl ester  with Retention Time (RT)=21.906


 

Figure 18: Mass spectrum of   Ergosta-5,22-dien-3-ol,acetate , ( 3β,22E)-  with Retention Time (RT)=23.119

 

 

Figure 19: Mass spectrum of   Campesterol  with Retention Time (RT)= 23.543

 

Figure 20: Mass spectrum of Cholest-5-en-3-ol ,24-propylidene-,(3β)-   with Retention Time (RT)=24.195


 

 


RESULTS AND DISCUSSION:

Identification of biochemical compounds

Analysis of compounds was carried out in methanolic extract of Lepidium sativum, shown in Table 1. The GC-MS chromatogram of the peaks of the compounds detected was shown in Figure 1. Chromatogram GC-MS analysis of the methanol extract of Lepidium sativum showed the presence of thirty one major peaks and the components corresponding to the peaks were determined as follows. All peaks were determined to be Glycerin , Monoethanolamine , 1-Deoxy-d-mannitol , 1-Nitro-2-propanol , 2-Butanamine , (S)- ,  Furfural , Allyl isothiocyanate , Paromomycin, 2-Hydroxy-2-(5-methylfuran-2-yl)1-phenylethanone, 3,6-Diazahomoadamantan-9-one Hydrazone, 2,3,4-Trimethoxycinnamic acid, 2-Naphthalenol, 2,3,4,4a,5,6,7-octahydro-1,4a-dimethyl-7-(2)-, cis-Vaccenic acid, 9-Octadecenamide , γ-Tocopherol , Phthalic acid , decyl oct-3-yl ester , Ergosta-5,22-dien-3-ol,acetate , ( 3β,22E)- , Campesterol and Cholest-5-en-3-ol ,24-propylidene-,(3β). Chromatography is the term used to describe a separation technique in which a mobile phase carrying a mixture is caused to move in contact with a selectively absorbent stationary phase. Flowers are also much prized by some invalids being palatable and beautiful root is used in secondary syphilis and tenesmus. Seeds are diuretic, alterative, tonic, aphrodisiac, carminative, galaetogogue, and emmenogogue. Gas chromatography has a very wide field of applications. But, its first and main area of use is in the separation and analysis of multi component mixtures such as essential oils, hydrocarbons and solvents. Intrinsically, with the use of the flame ionization detector and the electron capture detector (which have very high sensitivities) gas chromatography can quantitatively determine materials present at very low concentrations. It follows, that the second most important application area is in pollution studies, forensic work and general trace analysis. Because of its simplicity, sensitivity, and effectiveness in separating components of mixtures, gas chromatography is one of the most important tools in chemistry. It is widely used for quantitative and qualitative analysis of mixtures, for the purification of compounds, and for the determination of such thermo chemical constants as heats of solution and vaporization, vapor pressure, and activity coefficients 51-55. A knowledge of the chemical constituents of plants is desirable not only for the discovery of therapeutic agents, but also because such information may be of great value in disclosing new sources of economic phytocompounds for the synthesis of complex chemical substances and for discovering the actual significance of folkloric remedies. Higher plants as sources of bioactive compounds continue to play a dominant role in the maintenance of human health 55-65.

 

CONCLUSION:

In recent years GC-MS studies have been increasingly applied for the analysis of medicinal plants as this technique has proved to be a valuable method for the analysis of non-polar components and volatile essential oil, fatty acids, lipids and alkaloids.

 

ACKNOWLEDGEMENT:

I thank Dr. Amean A. Al-yasiri, College of Nursing, for valuable suggestions and encouragement.

 

REFERENCES:

1.          Mohammed GJ, Kadhim MJ, Hameed IH. Proteus species: Characterization and herbal antibacterial: A review. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(11): 1844-1854.

2.          Shireen SK, Hameed IH, Hamza LF. Acorus calamus: Parts used, insecticidal, anti-fungal, antitumour and anti-inflammatory activity: A review. International Journal of Pharmaceutical Quality Assurance. 2017; 8(3): 153-157.  

3.          Huda JA, Hameed IH, Hamza LF. Anethum graveolens: Physicochemical properties, medicinal uses, antimicrobial effects, antioxidant effect, anti-inflammatory and analgesic effects: A review. International Journal of Pharmaceutical Quality Assurance. 2017; 8(3): 88-91. 

4.          Altaee N, Kadhim MJ, Hameed IH. Detection of volatile compounds produced by pseudomonas aeruginosa isolated from UTI patients by gas chromatography-mass spectrometry. International Journal of Toxicological and Pharmacological Research. 2016; 8(6): 462-470.

5.          Hussein HM, Hameed IH, Ubaid JM. Analysis of the secondary metabolite products of Ammi majus and evaluation anti-insect activity. International Journal of Pharmacognosy and Phytochemical research. 2016; 8(8): 1192-1189.

6.          Hussein HM, Ubaid JM, Hameed IH. Inscticidal activity of methanolic seeds extract of Ricinus communis on adult of callosobruchus maculatus (coleopteran:brauchidae) and analysis of its phytochemical composition. International Journal of Pharmacognosy and Phytochemical research. 2016; 8(8): 1385-1397.

7.          Ubaid JM, Hussein HM, Hameed IH. Determination of bioactive chemical composition of Callosobruchus maculutus and investigation of its anti-fungal activity. International journal of Pharmcognosy and Phytochemical research. 2016; 8(8): 1293-1299.

8.          Ibraheam IA, Hussein HM, Hameed IH. Cyclamen persicum: Methanolic Extract Using Gas Chromatography-Mass Spectrometry (GC-MS) Technique. International Journal of Pharmaceutical Quality Assurance. 2017; 8(4); 200-213.

9.          Ibraheam IA, Hadi MY, Hameed IH. Analysis of Bioactive Compounds of Methanolic Leaves extract of Mentha pulegium Using Gas Chromatography-Mass Spectrometry (GC-MS) Technique. International Journal of Pharmaceutical Quality Assurance. 2017; 8(4); 174-182.

10.       Hadi MY, Hameed IH, Ibraheam IA. Ceratonia siliqua: Characterization, Pharmaceutical Products and Analysis of Bioactive Compounds: A Review. Research Journal of Pharmacy and Technology. 2017; 10(10): 3585-3589.

11.       Hadi MY, Hameed IH, Ibraheam IA. Mentha pulegium: Medicinal uses, Anti-Hepatic, Antibacterial, Antioxidant effect and Analysis of Bioactive Natural Compounds: A Review. Research Journal of Pharmacy and Technology. 2017; 10(10): 3580-3584.

12.       Kadhim MJ, Sosa AA, Hameed IH. Evaluation of anti-bacterial activity and bioactive chemical analysis of Ocimum basilicum using Fourier transform infrared (FT-IR) and gas chromatography-mass spectrometry (GC-MS) techniques. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(6): 127-146.

13.       Mohammed GJ, Kadhim MJ, Hussein HM. Characterization of bioactive chemical compounds from Aspergillus terreus and evaluation of antibacterial and antifungal activity. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(6): 889-905.  

14.       Hameed IH, Altameme HJ, Idan SA. Artemisia annua: Biochemical products analysis of methanolic aerial parts extract and anti-microbial capacity. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2016; 7(2): 1843- 1868

15.       Hussein AO, Mohammed GJ, Hadi MY, Hameed IH. Phytochemical screening of methanolic dried galls extract of Quercus infectoria using gas chromatography-mass spectrometry (GC-MS) and Fourier transform-infrared (FT-IR). Journal of Pharmacognosy and Phytotherapy. 2016; 8(3): 49-59.

16.       Sosa AA, Bagi SH, Hameed IH. Analysis of bioactive chemical compounds of Euphorbia lathyrus using gas chromatography-mass spectrometry and fourier-transform infrared spectroscopy. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(5): 109-126.

17.       Altameme HJ, Hadi MY, Hameed IH. Phytochemical analysis of Urtica dioica leaves by fourier-transform infrared spectroscopy and gas chromatography-mass spectrometry. Journal of Pharmacognosy and Phytotherapy. 2015; 7(10): 238-252.

18.       Mohammed GJ, Omran AM, Hussein HM. Antibacterial and Phytochemical Analysis of Piper nigrum using Gas Chromatography-Mass Spectrum and Fourier-Transform Infrared Spectroscopy. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(6): 977-996.

19.       Hamza LF, Kamal SA, Hameed IH. Determination of metabolites products by Penicillium expansum and evaluating antimicobial activity. Journal of Pharmacognosy and Phytotherapy. 2015; 7(9): 194-220.

20.       Jasim H, Hussein AO, Hameed IH, Kareem MA. Characterization of alkaloid constitution and evaluation of antimicrobial activity of Solanum nigrum using gas chromatography mass spectrometry (GC-MS). Journal of Pharmacognosy and Phytotherapy. 2015; 7(4): 56-72.

21.       Hadi MY, Mohammed GJ, Hameed IH. Analysis of bioactive chemical compounds of Nigella sativa using gas chromatography-mass spectrometry. Journal of Pharmacognosy and Phytotherapy. 2016; 8(2): 8-24.

22.       Hameed IH, Ibraheam IA, Kadhim HJ. Gas chromatography mass spectrum and fourier-transform infrared spectroscopy analysis of methanolic extract of Rosmarinus oficinalis leaves. Journal of Pharmacognosy and Phytotherapy. 2015; 7 (6): 90-106.

23.       Shareef HK, Muhammed HJ, Hussein HM, Hameed IH. Antibacterial effect of ginger (Zingiber officinale) roscoe and bioactive chemical analysis using gas chromatography mass spectrum. Oriental Journal of Chemistry. 2016; 32(2): 20-40.

24.       Al-Jassaci MJ, Mohammed GJ, Hameed IH. Secondary Metabolites Analysis of Saccharomyces cerievisiae and Evaluation of Antibacterial Activity. International Journal of Pharmaceutical and Clinical Research. 2016; 8(5): 304-315.

25.       Mohammed GJ, Al-Jassani MJ, Hameed IH. Anti-bacterial, Antifungal Activity and Chemical analysis of Punica grantanum (Pomegranate peel) using GC-MS and FTIR spectroscopy. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(3): 480-494.

26.       Dhahir BM, Hameed IH, Jaber AR. Prospective and Retrospective Study of Fractures According to Trauma Mechanism and Type of Bone Fracture. Research Journal of Pharmacy and Technology. 2017; 10(10):1827-1835.

27.       Hapeep MA, Hameed IH, Jasim AA. Risk Factors, Cause and Site of Firearm Injuries: A Prospective and Retrospective Study. Research Journal of Pharmacy and Technology. 2017; 10(10): 3420-3425.

28.       Jasim AA, Hameed IH, Hapeep MA. Traumatic Events in an Urban and Rural Population of Children, Adolescents and Adults in Babylon Governorate - Iraq. Research Journal of Pharmacy and Technology. 2017; 10(10): 3429-3434.

29.       Al-Marzoqi AH, Hadi MY, Hameed IH. Determination of metabolites products by Cassia angustifolia and evaluate antimicobial activity. Journal of Pharmacognosy and Phytotherapy. 2016; 8(2): 25-48.

30.       Altameme HJ, Hameed IH, Abu-Serag NA. Analysis of bioactive phytochemical compounds of two medicinal plants, Equisetum arvense and Alchemila valgaris seed using gas chromatography-mass spectrometry and fourier-transform infrared spectroscopy. Malays. Appl. Biol. 2015; 44(4): 47–58.

31.       Hameed IH, Hamza LF, Kamal SA. Analysis of bioactive chemical compounds of Aspergillus niger by using gas chromatography-mass spectrometry and fourier-transform infrared spectroscopy. Journal of Pharmacognosy and Phytotherapy. 2015;7(8): 132-163.

32.       Hameed IH, Hussein HJ, Kareem MA, Hamad NS. Identification of five newly described bioactive chemical compounds in methanolic extract of Mentha viridis by using gas chromatography-mass spectrometry (GC-MS). Journal of Pharmacognosy and Phytotherapy. 2015; 7 (7): 107-125.

33.       Hussein HM, Hameed IH, Ibraheem OA. Antimicrobial Activity and spectral chemical analysis of methanolic leaves extract of Adiantum Capillus-Veneris using GC-MS and FT-IR spectroscopy.  International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(3): 369-385.

34.       Hussein HJ, Hadi MY, Hameed IH. Study of chemical composition of Foeniculum vulgare using Fourier transform infrared spectrophotometer and gas chromatography - mass spectrometry. Journal of Pharmacognosy and Phytotherapy. 2016; 8(3): 60-89.

35.       Kadhim MJ, Mohammed GJ, Hameed IH. In vitro antibacterial, antifungal and phytochemical analysis of methanolic fruit extract of Cassia fistula. Oriental Journal of Chemistry. 2016; 32(2): 10-30.

36.       Altameme HJ, Hameed IH, Idan SA, Hadi MY. Biochemical analysis of Origanum vulgare seeds by fourier-transform infrared (FT-IR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Journal of Pharmacognosy and Phytotherapy. 2015; 7(9): 221-237.

37.       Hussein HM. Analysis of trace heavy metals and volatile chemical compounds of Lepidium sativum using atomic absorption spectroscopy, gas chromatography-mass spectrometric and fourier-transform infrared spectroscopy. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2016; 7(4): 2529 – 2555.

38.       Hameed IH. A new polymorphic positions discovered in mitochondrial DNA hypervariable region HVIII from central and north-central of Iraq. Mitochondrial DNA. 2016; 27(5): 3250-4.

39.       Jaddoa HH, Hameed IH, Mohammed GJ. Analysis of volatile metabolites released by Staphylococcus aureus using gas chromatography-Mass spectrometry and determination of its antifungal activity. Oriental Journal of Chemistry. 2016; 32(4): 8-24.

40.       Hameed IH, Salman HD, Mohammed GJ. Evaluation of antifungal and antibacterial activity and analysis of bioactive phytochemical compounds of Cinnamomum zeylanicum (Cinnamon bark) using gas chromatography-mass spectrometry. Oriental Journal of Chemistry. 2016; 32(4): 16-25.

41.       Hameed IH, Jebor MA, Ommer AJ, Abdulzahra AI. Haplotype data of mitochondrial DNA coding region encompassing nucleotide positions 11,719–12,184 and evaluate the importance of these positions for forensic genetic purposes in Iraq. Mitochondrial DNA. 2016; 27(2): 1324-1327.

42.       Kadhim MJ, Mohammed GJ, Hussein HM. Analysis of bioactive metabolites from Candida albicans using (GC-MS) and evaluation of antibacterial activity. International Journal of Pharmaceutical and Clinical Research. 2016; 8(7): 655-670.

43.       Mohammad A, Imad H. Autosomal STR: From locus information to next generation sequencing technology. Research Journal of Biotechnology. 2013; 1(2): 15-23. 

44.       Hameed IH, Abdulzahra AI, Jebor MA, Kqueen CY. Ommer, A.J. Haplotypes and variable position detection in the mitochondrial DNA coding region encompassing nucleotide positions 10,716-11,184. Mitochondrial DNA. 2015; 3(6): 1-9.

45.       Ubaid JM, Hussein HM, Hameed IH. Analysis of bioactive compounds of Tribolium castaneum and evaluation of anti-bacterial activity. International Journal of Pharmaceutical and Clinical Research. 2016; 8(7): 655-670.

46.       Altaee N, Kadhim MJ, Hameed IH. Detection of volatile compounds produced by Pseudomonas aeruginosa isolated from UTI patients by gas chromatography-mass spectrometry. International Journal of Current Pharmaceutical Review and Research. 2017; 7(6): 8-24.

47.       Altaee N, Kadhim MJ, Hameed IH. Characterization of metabolites produced by E. coli and analysis of its chemical compounds using GC-MS. International Journal of Current Pharmaceutical Review and Research. 2017; 7(6): 13-19.

48.       Hussein JH, Ubaid JM, Hameed IH. Gas chromatography – mass spectrum analysis of volatile components of methanolic leaves extract of Cordia myxa. International Journal of Current Pharmaceutical Review and Research. 2017; 7(6): 16-22.

49.       Hameed, I.H., Al-Rubaye A.F. and Kadhim, M.J. Antimicrobial Activity of Medicinal Plants and Urinary Tract Infections. International Journal of Pharmaceutical and Clinical Research. 2017; 8(11): 44-54. 

50.       Kadhim WA, Kadhim, M.J., Hameed, I.H. Antibacterial Activity of Several Plant Extracts Against Proteus Species. International Journal of Pharmaceutical and Clinical Research. 2017; 8(11): 88-94.

51.       Ahmed MD, Hameed IH, Abd-Ali MQ. Prospective and Retrospective Study of the Acute Heart Attack Cases in Marjan Hospital-Hillah City-Iraq. Research Journal of Pharmacy and Technology. 2017; 10(10): 3408-3416.

52.       Fakhir DF, Hameed IH, Flayyih SS. Burns Injuries: A Prospective Statistical Study of 112 patients. Research Journal of Pharmacy and Technology. 2017; 10(10): 3401-3407.

53.       Mekhlef AK, Hameed IH, Khudhair ME. Prevalence of Physical Injuries on the Head, Neck and Entire Body in, Hilla, Iraq. Research Journal of Pharmacy and Technology. 2017; 10(10): 3276-3282.

54.       Kadhim MJ. In Vitro antifungal potential of Acinetobacter baumannii and determination of its chemical composition by gas chromatography-mass spectrometry. Der Pharma Chemica. 2016; 8(19): 657-665.

55.       Hameed IH, Al-Rubaye AF, Kadhim MJ. Antimicrobial Activity of Medicinal Plants and Urinary Tract Infections. International Journal of Pharmaceutical and Clinical Research. 2017; 9(1): 44-50.

56.       Al-Rubaye AF, Hameed IH, Kadhim MJ. A Review: Uses of Gas Chromatography-Mass Spectrometry (GC-MS) Technique for Analysis of Bioactive Natural Compounds of Some Plants. International Journal of Toxicological and Pharmacological Research. 2017; 9(1); 81-85.

57.       Al-Rubaye AF, Kadhim MJ, Hameed IH. Characterization of Antifungal Secondary Metabolites Produced by Klebsiella pneumoniae and Screening of its Chemical Compounds Using GC-MS. International Journal of Current Pharmaceutical Review and Research. 8(2); 141-148.

58.       Hameed IH, Al-Rubaye AF, Kadhim MJ. Uses of Nuclear Magnetic Resonance Spectroscopy Technique in Pharmaceutical Analysis: A Review. International Journal of Current Pharmaceutical Review and Research. 2017; 8(2): 79-84.

59.       Al-Rubaye AF, Kadhim MJ, Hameed IH. Phytochemical Profiles of Methanolic Seeds Extract of Cuminum cyminum using GC-MS Technique. International Journal of Current Pharmaceutical Review and Research. 2017; 8(2): 114-124.

60.       Hameed IH, Al-Rubaye AF, Kadhim MJ. Urinary Tract Infections: Characterization and Herbal Antimicrobial Activity: A Review. International Journal of Current Pharmaceutical Review and Research. 2017; 8(2): 184-191.

61.       Kadhim MJ, Kaizal AF, Hameed IH. Medicinal Plants Used for Treatment of Rheumatoid Arthritis: A Review. International Journal of Pharmaceutical and Clinical Research. 2016; 8(12): 1685-1694.

62.       Ubaid JM, Kadhim MJ, Hameed IH. Study of Bioactive Methanolic Extract of Camponotus fellah Using Gas Chromatography – Mass Spectrum. International Journal of Toxicological and Pharmacological Research. 2016; 8(6): 434-439.

63.       Hussein HM, Hameed IH, Ubaid JM. Analysis of the secondary metabolite products of Ammi majus and evaluation anti-insect activity. International journal of Pharmacognosy and Phytochemical Research. 2016; 8(8): 1192-1189.

64.       Hussein HM, Ubaid JM, Hameed IH. Inscticidal activity of methanolic seeds extract of Ricinus communis on adult of callosobruchus maculatus (coleopteran:brauchidae) and analysis of its phytochemical composition. International Journal of Pharmacognosy and Phytochemical Research. 2016; 8(8): 1385-1397.

65.       Ubaid JM, Hussein HM, Hameed IH. Determination of bioactive chemical composition of Callosobruchus maculutus and investigation of its anti-fungal activity. International Journal of Pharmcognosy and Phytochemical research. 2016; 8(8): 1293-1299.

 

 

 

Received on 05.07.2017             Modified on 22.08.2017

Accepted on 28.09.2017           © RJPT All right reserved

Research J. Pharm. and Tech 2017; 10(11): 3981-3989.

DOI: 10.5958/0974-360X.2017.00723.5