Showing 6 results for Chromatography
Sabzevari O, Daliraj A, Mohammadi M, Rastegar H,
Volume 64, Issue 12 (11-2006)
Abstract
Background: Infant formula, depending on the source, contains various fatty acids, which may possess important nutritional and biological value for infants. The presence of some of these fatty acids in infant formula, however, can be harmful and toxic for the infant. In this regard, more attention has been paid to erucic acid since its accumulation in myocardial tissues may cause damage to the heart. Therefore, a limit has been set by the Codex Alimentarius for the presence of erucic acid in infant formula (less than 1% of total fatty acids). The purpose of the present study is to investigate amount of erucic acid present in three infant formulas used predominantly in Iran.
Methods: Gas chromatography (GC) is a valuable method applied for the separation of fatty acids, including erucic acid, from oils and oily food. Three brands of infant formulas, namely Humana, Biomil and Multi, were analyzed by GC using a wall coated open tubular (WCOT) fused silica column and flame ionization detector (FID). Heneicosanoic acid was employed as an internal standard.
Results: The findings showed that Humana and Biomil infant formula samples contained 0.06% and 0.002% erucic acid (from total fatty acids), respectively, while no erucic acid was detected in the Multi infant formula samples.
Conclusion: The amount of erucic acid in the studied infant formulas was far below the Codex limit of 1% total fatty acids.
Mahmoodzadeh A, Morady A, Zarrinnahad H, Pooshang Bagheri K, Ghasemi-Dehkordi P, Mahdavi M, Shahbazzadeh D, Shahmorady H,
Volume 70, Issue 12 (3-2013)
Abstract
Background: Gastric cancer (GC) is one of the most common cancers worldwide and in Iran. Conventional therapies are surgery and chemotherapy. Current studies are evaluating natural compounds in inhibiting growth of cancer cell. In this study isolated peptide melittin with 26 amino acids from bee venom and its impact on the viability and proliferation of gastric cancer cells was investigated.
Methods: At first melittin was purified from honeybee venom using a reversed-phase high performance liquid chromatography (RP- HPLC) and C18 column. In order to investigate whether melittin, a 26 amino acids peptide which is the main components of honeybee venom, inhibits proliferation of human gastric adenocarcinoma cell line (AGS cells), MTT ((3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide) assay was performed. Hemolytic assay carried out in order to confirm the biologic activity of the isolated melittin. AGS cells were plated in a 96-well plate and treated with serially diluted concentrations of melittin for 6 and 12 hours. The mortality of the cells was measured via MTT assay at 540 nm.
Results: The obtained chromatogram from RP-HPLC showed that melittin comprises 50% of the studied bee venom. SDS-PAGE analysis of melittin fraction confirmed purity of isolated melittin. Hemolytic activity assay indicates that isolated melittin shows a strong hemolytic activity (HD50=0.5). MTT assay showed that melittin strongly inhibits proliferation of gastric cancer cells at concentrations more than 2µg/ml. This inhibitory effect is dependent to melittin concentration and incubation time.
Conclusion: This study provides evidence that melittin inhibits proliferation of the gastric cancer cells. Results showed that isolated melittin from honey bee venom have cytotoxic effect on AGS cell line with a trend of increasing cytotoxicity with increasing concentration and incubation time.
Marjan Rismanchi , Pooneh Mokarram , Mahvash Alizadeh Naeeni , Mahdi Paryan , Zohreh Honardar , Soudabeh Kavousipour , Abbas Alipour ,
Volume 71, Issue 12 (3-2014)
Abstract
Background: Colorectal Cancer (CRC) is the third common cancer in the world. One of the pathways in colorectal tumor genesis is Microsatellite Instability (MSI+). MSI is detected in about 15% of all colorectal cancers. Colorectal tumors with MSI have dis-tinctive features compared with Microsatellite Stable (MSS) tumors. Due to the high percentage of MSI+ in patients with CRC in Iran, screening of this type of CRC is im-perative. In current study, two markers (BAT-26 and BAT-25) were used to determine an appropriate screening technique with high sensitivity and specificity to diagnose MSI status in patients with CRC.
Methods: Allelic variation in two markers (BAT-26 and BAT-25) was analyzed in tis-sues and sera of 44 normal volunteers and tumor and matched normal mucosal tissues as well as sera of 44 patients with sporadic colorectal cancer by Real Time PCR (Hy-bridization probe) and High-Performance Liquid Chromatography (HPLC) techniques. The sensitivity and specificity of Real Time PCR and HPLC compared with sequencing as gold standard. The data were statistically analyzed using Student’s t-test and 2 or fisher exact test, where applicable with (P<0.05). Receiver-operating-characteristic (ROC) curves were used to evaluate the sensitivity and specificity.
Results: The sensitivity and specificity of BAT-26 with Real Time PCR method (Hy-bridization probe) were 100% in comparison with gold standard method. Whereas the sensitivity and specificity of BAT-26 and BAT-25 with HPLC were 83%, 100% and 50%, 97%, respectively. Neither HPLC nor Real time PCR could detect circulating DNA with MSI property in sera.
Conclusion: The sensitivity and specificity of real time PCR in MSI detection is the same as sequencing method and more than HPLC. BAT-26 marker is more sensitive than BAT-25 and MSI detection with Real time PCR could be considered as an accu-rate method to diagnose MSI in CRC tissues not sera.
Alireza Ebrahimi , Zohre Niknami , Fahime Nazari , Mahasti Ghavami Adel , Amir Atashi , Abdolfattah Sarrafnejad ,
Volume 74, Issue 3 (6-2016)
Abstract
Hemoglobinopathies are most common inherited disorders in the world; approximately 7 percent of the worldwide population and 5-6 percent of population of Iran are carriers. The hemoglobin disorders inherit as autosomal recessive and are very common in the Mediterranean area and much of the Asia and Africa. The control of this inherited disorders need to genetic counseling and accurate screening by more advanced and more accurate methods. This study explains features of current Iran hemoglobin disorders, nominates the accessible methods for screening them and introduces the capillary zone electrophoresis as a rapid and more accurate method. The required data were extracted of various articles and then for good explanation, current Iran hemoglobinopathies properties were showed in the tables and electropherograms of important hemoglobin disorders in Iran population were provided for help to interpretation results of blood tests by capillary zone electrophoresis method. Hemoglobin disorders are including thalassemias and hemoglobin variants; Disruption in the production and malfunction of globin chains cause types of hemoglobin disorders. We cannot introduce one of clinical laboratory tests as critical and basic method for screening and distinguishing types of inherited hemoglobin disorders as alone. For distinguishing the types of them must be prepared enough information and data of the hemoglobin disorders and for more accurate analysis must be used simultaneously different methods as gel electrophoresis, high performance liquid chromatography, isoelectric focusing, capillary zone electrophoresis or molecular tests. The capillary electrophoresis is an accurate and rapid method for screening types of the hemoglobin disorders. Other side this method cannot analyze all of them, so must be used biochemical, biophysical and molecular methods for confirmation the results. This review showed we can use the capillary electrophoresis and HPLC as two complementary methods for hemoglobinopathies screening. We can analyze by the methods more hemoglobin disorders and decrease more laboratory errors. Moreover, we must have patient history, hematological indices, information and data of types of hemoglobinopathies. The patient history and complete blood count results as red blood cell count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration can be useful and helpful in screening the hemoglobin disorders and then distinguishing all of hemoglobin disorders.
Amir Mirzaie , Shoreh Zare Karizi ,
Volume 74, Issue 9 (12-2016)
Abstract
Background: Centaurea cyanus is an endemic and well-known herbal medicine in Iran, is an annual flowering plant in the family of Asteraceae. The flowers are the part used in modern herbal medicine and are considered to have tonic, stimulant and emmenagogue properties, with action similar to that of blessed thistle. The aim this study was to investigate the phytochemical constituents of C. cyanus extract, its antioxidant, anti-tumor and anti-bacterial activities.
Methods: This experimental study was conducted from June to January of 2015 in Islamic Azad University of Varamin, Iran. At first, the phytochemical components of C. cyanus extract was analyzed using gas chromatography–mass spectrometry (GC-MS) method. Subsequently, the antibacterial potential of the extract was evaluated against 4 pathogenic bacteria including Staphylococcus aureus, Streptococcus pyogenes, Psedomonas aeroginosa and Klebsiella pnemoniae via minimum inhibitory concentration (MIC) mathod. Moreover, the anti-oxidant and anti-tumor activities of extract on colon cancer cell line (HT29) were investigate using DPPH and MTT colorimetric methods, respectively. Finally, the Bax and Bcl2 apoptosis gene expression level was analyzed by quantitative Real-time PCR technique.
Results: GC-MS analysis of C. cyanus extract was shown 19 major components and the most frequent component was belonged to n-Hexadecanoic acid (36.4%) and Linoleic acid (19.3%). The maximum antibacterial activity of extract was observed on S. aureus and P. aeroginosa isolates. The antioxidant activity of the extract was 0.109±0.07 mg/ml. Moreover, the MTT results show that extract had IC50= 26.04±0.45 on HT29 cell line. The Real-time PCR results showed the expression level of Bax and Bcl2 was significantly increased and decreased respectively in colon cancer cell line (2.63±0.54 (P< 0.05), 0.38±0.72 (P< 0.05)).
Conclusion: The results of this study show that the extract had significant anti-bacterial and anti-cancer effects and it appear that the extract has potential uses for pharmaceutical industries.
Mariam Bagheri, Hashem Khorsand Mohammadpour, Kamran Mousavi Hosseini ,
Volume 78, Issue 12 (3-2021)
Abstract
Background: Due to multiple roles of albumin in the body, injection of its medicinal product as one of the therapeutic or management strategies under conditions such as severe bleeding, burns, liver failure, and neonatal hemolytic diseases is on the physicians' agenda. Considering that albumin is the most abundant plasma protein, designing an appropriate method to purify it is highly important. There are several methods such as human plasma fractionation, chromatographic, or Salting-out methods for the isolation and purification of the human albumin. The present study investigates a direct and combined ion-exchange chromatography approach for purification of albumin from human plasma and compares the quality of the final products obtained by both ion-exchange chromatographic methods.
Methods: This study was carried out from January 2019 to October 2019 at the Blood Transfusion Research Center, High institute for research and education in transfusion medicine, affiliated with the Iranian Blood Transfusion Organization. For this study, 10 human plasma bags were randomly collected. After thawing, all 10 human plasma bags were pooled, and in order to separate cryo paste, it was centrifuged at 4000 g for 10 minutes at the temperature of 1 Centigrade degree. Then the obtained cryo poor plasma was used to purify the albumin protein by direct and combined methods of ion-exchange chromatography. The purity of the final products was compared by cellulose acetate electrophoresis and SDS-PAGE tests. The sample obtained by the combined approach was pasteurized and HPLC analysis was performed to investigate any polymer aggregates.
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Results: In contrast to the direct method, the final product obtained by combined ion-exchange chromatography had a good purity by the average of about 95% and the amount of polymer was estimated to be less than 5% by HPLC analysis (P<0.05).
Conclusion: By diluting the plasma and subsequently reducing the ionic strength, albumin can be separated from human plasma with a high degree of purity only by two steps of ion-exchange chromatography.
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