Vol. 31, November, 2009.


Amany A. Salem and Naglaa H. M. Hassanen

Food Technology Research Institute, Agricultural Researches Center, Giza, Egypt.

The current study is an alternative treatment using Ammi visnaga L. “khella” on nephrocalcinsis rats as a preventive agent against the development of kidney stones. The experimental was conducted in normal and nephrocalcinosis rats during 40 days. Several parameters were followed after 10, 25 and 40 days as three stages of experimental period. The parameters examined included weight of the rats at the end each of stage, water intake, urinary volume, liver functions in serum, kidney functions in serum and urine and some minerals in serum and urine. At the end of each stage, rats were killed followed by weight of organs and histopathological was studied. Also, the present study deigned to produce some products by addition Ammi visnaga L. seeds as especial foods for kidney patients which were sensory evaluated. The results show that, there are non significant differences between groups for organs weight except that of kidney of the nephrocalcinosis rats which had the highest weight compared with control rats. No significant difference between groups for liver functions. The treatments with Ammi visnaga L. show that, the water extracts were improved the kidney functions and tissues more than seeds. Also, the whole seeds were better than the ground seeds for kidney functions, tissues and urinary chemistries. Histopathological showed a slight differences in liver tissues. But, the kidney tissues resulted in, interstitial fibrosis, associated with cystic dilatation, marked vacuolations of epithelial lining and necrosis of renal tubules. The sensory evaluation showed that, the addition of water extracts to dibis, fig jam and arake juice changed the colour of product and had the lowest scores for taste.  




Omneya Khowessah, Maha Amin and Emad B. Basalious

Department of Pharmaceuticsand Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Kasr El Aini Street, Egypt.

Aceclofenac-loaded PLGA nanoparticles were prepared by spontaneous emulsification solvent diffusion method (SESD). The composition of the organic phase was varied to study the effect of acetone to dichloromethane ratio on nanoparticle formation, particle size and entrapment efficiency. Results indicated that 1:1 ratio of acetone:dichloromethane was the optimum organic solvent mixture for nanoparticle preparation. A 24 full factorial design based on the selected organic solvent mixture was applied. The independent variables were the amount of drug, the concentration of PVA in the aqueous phase as well as the type of PLGA and its concentration. The effects of these parameters on entrapment efficiency, particle size and drug release were investigated. Differential scanning calorimetric (DSC) studies and measurement of zeta potential were performed on the optimized aceclofenac-loaded nanoparticles formulation to investigate the crystalline nature of the drug after entrapment and the surface charge of the prepared nanoparticles. Results revealed that increasing PLGA concentration results in higher entrapment efficiency, while the use of 5% PVA in the aqueous external phase results in smaller particle size of the prepared nanoparticles. In-vitro release studies in phosphate buffer (pH 7.4) showed extended drug release and fitted the theoretical target release profile. Most formulations exhibited Fickian diffusion drug release profiles. The optimized formulation, prepared using 94.07 mg aceclofenac, 400mg PLGA-RH and 4.72 % PVA, had entrapment efficiency of 67.72 %, a mean particle size of 593 nm and exhibited a prolonged release (t50%=5.23hrs). Aceclofenac loaded PLGA nanoparticles seem to be a promising ocular delivery system for extending the ocular anti-inflammatory action of aceclofenac.




Wafaa M.E. Abdel Fattah* Hoda M. El-Kholy* and Sherif A. Nassib**

*Biochemistry Department and **Internal Medicine Department, Faculty of Medicine for Girls, Al-Azhar University

Background and aim of work: Immunomodulatory mediators play a crucial role in the pathogenesis of heart failure [HF]. Vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-l (ICAM-1) are important inflammatory mediators of leukocyte adhesion to vascular endothelium. The plasma level of the soluble forms of these molecules elevate in chronic and acute inflammation. Furthermore, the immune modulator CD40-CD40 ligand (CD40L) has been receiving increased attention, since it plays a key role in the pathophysiology of multicellular vascular events such as thrombosis, inflammation, and atherosclerosis. Based on the previous fact we sought to evaluate the role of plasma VCAM-1, ICAM-l and soluble CD40L in human HF. Subjects and methods: This study was done on 30 male patients their ages ranged from 45 to 55 years. Fifteen patients have previous history of ischemic heart with atherosclerosis and are suffering from chronic HF and fifteen patients with acute myocardial infarction (MI) complicated with HF during acute phase, with past history of hypertension. Another 10 normal healthy male volunteers were taken as control with the same age range. Plasma VCAM-1, ICAM-1 and serum soluble CD40L [sCD40L] levels were measured in all groups. Lipid profile, serum creatine phosphokinase (CPK) and its (MB) fraction were also measured for patients only. Our results revealed that there were significant increase in plasma VCAM-1, ICAM-1 and serum sCD40L. In both chronic and acute heart failure cases compared to control P<0.01. Our results also revealed that a positive significant correlation between sCD40L and VCAM-1 in chronic HF r=0.565, P= 0.028. and a positive significant correlation between sCD40L and ICAM-1 in acute heart failure r=0.851, P<0.01 also there was a positive significant correlation between sCD40L and LDL in both chronic and acute HF cases P<0.01.



Abd El-Salam I. Mohammed

Department of Pharmacognosy, Faculty of Pharmacy, Al-Azhar University,

Cairo, Egypt

Three anthraquinone aglycons; chrysophanol, physcion and emodin, together with two anthraquinone glycosides; emodin-6-O-α-L-rhamnopyranoside and emodin-6-O-β-D-glucopyranoside were isolated for the first time from the stem of Rhamnus disperma. The structure identification of the isolated glycosides have been established on the bases of NMR spectroscopic data. Whereas,the aglycons have been established by co-TLC with authentic samples as well as EI-MS.


Esmat E. Zein El-Din*, Sanna A. El-Gizawy*, Sahar M. El-Haggar**,

and Doaa A. Habib*

*Department of Pharmaceutical Technology and **Department of Clinical Pharmacy, Faculty of Pharmacy, Tanta University, Tanta, Egypt.

The purpose of this research was to prepare meloxicam tablets with enhanced in vitro dissolution utilizing different formulation additives.For poorly soluble, highly permeable (class II) drugs, such as meloxicam, the rate of oral absorption is often controlled by the dissolution rate in the gastrointestinal tract. Different formulations of meloxicam tablets were prepared by applying a 23 full factorial design to investigate the influences of three independent variables on the in vitro drug dissolution: type of diluent (X1), type of disintegrant (X2) and inclusion of alkalinizing agent (X3). The results indicated that factor X1 showed the largest significant effect on drug dissolution followed by factor X3 in pH 6.8. In pH 1.0, factor X3showed the highest coefficient value followed by factor X1.Meloxicam tablets R, F3 and F6 were considered for in vivo study representing the reference product, and the tablets of lowest and highest dissolution, respectively. Single dose oral pharmacokinetic study in healthy adult volunteers revealed that F6 showed significantly higher (P<0.05) absorption rate (Ka) compared with R and F3. In conclusion, the formulation additives may play a significant role in improving in vitro dissolution and consequently in vivo absorption of class II drugs.




Manal M. Fouad, *Afaf O. Mohamed,   Sawsan A. Abdel Razeq, *Zeinab A.Elsherif

and *Mona M. Hasan

Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University,

Cairo, Egypt

*National Organization For Drug Control and Research (NODCAR), Giza, Egypt

Three Simple, rapid and accurate methods are presented for the quantitative determination of tazarotene. The first is an HPLC method, performed on an Eclipse C18 column, using a mobile phase of acetonitrile-methanol-H2O (60:30:10 v/v/v) with UV detection at 353 nm. The second is based on the quantitative densitometric evaluation of thin layer chromatogram of tazarotene at 260 and 353 nm after developing with a mobile phase of toluene-chloroform-methanol-ammonia (5:6:3:0.1 v/v/v/v), whilst the third one measures the fluorescence intensity of tazarotene in acetone at 466 nm (λex 345 nm). Regression analysis of Beer’s plots show good correlations (0.9992 - 0.9996) over concentration ranges of 1-10 μg ml-1, 0.5 - 4.5 μg / spot and 0.05 - 0.5 μg ml-l for the three methods, respectively. These methods can be used as stability indicating ones to determine the intact drug in presence of its alkaline induced degradation products up to more than 90% for the first and second methods while up to 30% for the third one. The proposed methods are successfully applied to analyse tazarotene in its formulations; Acnitaz gel with mean recoveries ranging from 98.22 to 99.78 %. The results obtained are validated and statistically analysed and found to be in accordance with those given by a reported method.



Rafat M. Mohareb, Karam A. El-Sharkaway, *Mona M. Hussein

and **Hend M. El-Sehrawi

Organic Chemistry Department, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), El-Wahat Road, 6 October City, Egypt.

*Department of Chemistry, Faculty of Science, Cairo University, Giza, A. R. Egypt.

**Pharmaceutical Chemistry Department, Faculty of Pharmacy (Girls), El-Azhar University, Nasr City, Cairo, A. R. Egypt.

The reaction of cyanoacetylhydrazine (I) with w-bromo(4-methoxyacetophenone (II) gave the hydrazide-hydrazone derivative III. Compound III reacted with either potassium cyanide or potassium thiocyanide to give the cyanide or thiocyanide derivatives IVa or IVb respectively. The reaction of compound III with either hydrazine hydrate or phenylhydrazine gave the hydrazine derivatives VIa or VIb respectively. The latter compounds underwent a series of heterocyclization when react with different reagents to give 1,3,4-triazine and pyridine derivatives. The antidepressant, sedative and analgesic activities of the newly synthesized products were evaluated.



Sahar M. El-Haggar

Department of Clinical Pharmacy, Faculty of Pharmacy, University of Tanta,

Tanta, Egypt

The purpose of this research was to utilize the design of experiment in optimization of topical delivery of sildenafil from gel formulations. Sildenafil is the first oral therapeutic agent used for the management of male erectile dysfunction. Topical delivery of sildenafil may be a successful alternative to oral administration. This is because topical drug delivery avoids pre-systemic metabolism, may sustain plasma drug levels to overcome short duration of action and may reduce incidence of adverse effects. Different formulations of sildenafil gel were prepared by applying a 22 full factorial design to investigate the influences of two independent variables on the in vitro drug release: the type of enhancers (A) at two levels (10% PEG 400 and 10% Tween 80) and type of gelling agent (B) at two levels (3% HPMC and 20% Poloxamer). All the designed formulations were subjected to in vitro and in vivo evaluations. The release rate constant (k), the consistency index (K), thixotropic index, and the degree of erectile erection were selected as dependent variables. The drug release pattern of all the tested formulations was best fitted to zero-order. The rheological behavior was pseudo-plastic for all designed formulations. The statistical analysis indicated that factors A and B have their largest significant effect on release rat constant and consistency index in their low level (Poloxamer) and high level (PEG 400), respectively. Factor B has higher coefficient value compared with A indicating that more significant role of the gelling agent in improving drug release. The clinical study was conducted by applying the gel topically on the glans of the penis of 40 patients with erectile dysfunction (ED) in a random, parallel, double blind, prospective and placebo-controlled design. The highest degree of erection was achieved by the optimized formula in relation to drug release and viscosity and containing Poloxamer and PEG 400. In conclusion, the optimized formula may be considered as a promising tool for topical drug delivery for treatment of ED.




Wafaa M.E. Abdel Fattah*, Hani A. Hussein** and Maha Abou-Zekri***

*Biochemistry Department, Faculty of Medicine for Girls, Al-Azhar University.

** Pediatrics Department, Faculty of Medicine for Boys, Al-Azhar University and ***Pediatrics Department, Kasr-Al Ainy Faculty of Medicine, Cairo University.

Background: Helicobacter pylori (H. pylori) infections are relatively common infection and worldwide in distribution. The causative organism is Helicobacter pylori which is a gram negative, urease producing bacillus. H.pylori infection has been correlated with iron deficiency anemia mainly in children. Iron deficiency might affect oxygen metabolism, free radicals and DNA synthesis through cytochromic enzymes and other metalloenzymes. Iron deficiency anemia is also associated with fall in serum ferritin which is the major iron storage protein in nearly all organisms. Oxidative stress and oxidative damage caused by free radicals is believed to play a role in pathogensis of H.pylori infection. This oxidative stress cause lipid peroxidation and protein glycation can be assessed by measurement of Malondialdhyde (MDA) and Fructosamine respectively. Aim of work: Given the importance of glycated protein in causing pathological complication in various diseases, it was deemed pertinent to investigate the levels of glycated plasma protein and the possible association with the levels of lipid peroxides in H. pylori infected patients who were anemic. Furthermore, to explore the effect of treatment on the levels of these parameters. Subjects and methods: This study was done on 30 children (17 females, 13 males). They were divided into two groups. The first group contains 15 healthy children were taken as a control and the second group contains 15 H.pylori infected anemic children. Blood samples were collected from both groups. The infected group then received anti-H.pylori therapy for 14 days and iron supplementation for six weeks and another blood samples were collected. Serum MDA, fructosamine and glucose were evaluated in control and in patients before and after treatment. Serum iron, ferritin and hemoglobin were also evaluated in both groups. Our results revealed a highly significant increase in the levels of MDA and fructosamine in anemic H.pylori infected children compared with control group (P<0.01) while iron,hemoglobin and ferritin levels were highly significantly decreased.In anemic H.pylori group receiving anti H.pylori therapy and iron supplement,the result show MDA and fructosamine levels decreased significantly (P<0.01) and (P<0.05) respectively compared to the same group before therapy. There was a significant increase in iron (P<0.01),hemoglobin (P<0.01) and ferritin (P<0.01) levels after six weeks of treatment.




Manal M. Fouad,Sawsan A. Abdel Razeq, Zeinab A. Elsaid, *Lobna A. Hussin

and *Amira M. Elkosasy

Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University,

Cairo, Egypt.

*Analytical Chemistry Department, Faculty of Pharmacy, Ein-Shams University, Cairo, Egypt.

Two simple, sensitive and valid methods for the determination of nalbuphine- hydrochloride are investigated. The first is a spectrophotometric method involving the use of three reagents, where nalbuphine HCl reacts with 3- methyl–2–benzothiazolinone in presence of Ce (IV) via an oxidative coupling mechanism to yield a red colored product measured at 420 and 536 nm. Owing to its reducing properties, the drug also reacts with Folin–ciocaltea’s reagent or phenanthroline– iron (III) to produce a blue colored product or an orange–red ferrion complex measured at 730 nm or 513 nm, respectively. The second method is a spectrofluorimertic one in which the fluorescence intensity of the drug in water is measured at λem 323 nm (λex257 nm). The linear concentration ranges are 10–100, 5–40, 0.5–5 µg ml-1 for the spectrophotometric method regarding the three reagents, respectively and 1-10µg ml-1 for the fluorimetric method. The accuracies of the proposed methods amount to 98.4-100.8±0.13-1.21 % while inter and intraday RSD obtained are 0.05-2.63%. Optimum experimental conditions and their applicability to the determination of the cited drug in pharmaceutical formulations are described. The results obtained were statistically analyzed and found to be in accordance with those obtained from a reported method.




Magdy A. Al-Tahhan*, Samy H. Mohammad* and Ehab M. Salem**

Medical Biochemistry* and Internal Medicine** Departments, Faculty of Medicine, Zagazig University.

Diabetic nephropathy is a common complication in patients with either type 1 or type 2 diabetes. It is characterized by glomerulosclerosis and tubulointerstitial fibrosis as a result of overproduction and/or decreased degradation of extracellular matrix (ECM) in the kidney. In that context plasminogen activator inhibitor-1 (PAI-1) and visfatin may contribute to these processes .Twenty type 2 diabetic patients without nephropathy and 37 diabetic patients with nephropathy (17 with microalbuminuria and 20 with overt albuminuria) were recruited, matched for their age, sex and body mass index (BMI). Twenty healthy non diabetic subjects served as controls. Plasma PAI-1, visfatin and serum oxidation susceptibility of lipoproteins were measured for all subjects. Significantly higher plasma PAI-1 and visfatin levels were observed in all diabetic patients as compared to control group. Patients with nephropathy showed significantly higher mean plasma PAI-1 and visfatin levels compared to patients without nephropathy. Interestingly, patients with overt albuminuria showed significantly higher mean plasma PAI-1 and visfatin levels as compared with patients with microalbuminuria indicating that PAI-1 and visfatin are involved in the pathogenesis and in progression of diabetic nephropathy. Significant positive correlations were observed between blood glucose, HbA1c, serum triglycerides, cholesterol, oxidation susceptibility of lipoproteins and PAI-1 and visfatin levels, while significant negative correlation were detected between HDL-cholesterol and plasma PAI-1 and visfatin levels. In conclusion PAI-1 and visfatin could play a pivotal role in the pathogenesis and progression of the disease and interruption of PAI-1 and visfatin production by either therapeutic modulation or tight glycemic and lipidemic control might be useful in treatment and prevention of diabetic nephropathy.




Maha A. Marzouk and Zeinab M. Gebaly*

Department of Pharmaceutics, Faculty of Pharmacy, Department of Histology*, Faculty of Medicine. Al-Azhar University, Nasr City, Cairo, Egypt.

The objective of this study was to a design transdermal matrix system capable of diffusing Diclofenac sodium (DS) a non-steroidal anti-inflammatory drug, at a constant rate over an extended period of time as an alternative route of topical administration. The drug was characterized for its physicochemical properties like solubility in water and in buffer solutions of different pHs and partition coefficient using n-octanol. Transdermal patches containing DS were formulated using hydroxypropyl methylcellulose (HPMC) as polymer and were plasticized with polyethylene glycol 400 (PEG 400). The patches were evaluated for uniformity of thickness and weight, moisture content, moisture uptake, flatness and folding endurance. The in-vitro release rates of DS from HPMC patches containing different penetration enhancers, such as methyl nicotinate, camphor, glycrrhizin, thymol, dimethyl formamide (DMF), n-octanol, isopropyl myristate (IPM), propylene glycol (PG), α-tocopherol, turpentine oil, eucalyptus oil and methyl salicylate were studied. In-vitro skin permeation study using abdominal rabbit skin was conducted in a modified diffusion cell. The cumulative amount permeated at 24 h (µg/cm2), steady-state flux jss (µg/cm2. h), lag time tL(h), permeability coefficient kp (cm/h), partition coefficient k, and Diffusion coefficient D (cm2/h) were determined for the prepared patches in comparison with the commercial gel. An accelerated stability study was conducted for all the patches at 42oC and 75% RH for 6 months. The patches that gave best results, dealing with in-vitro release and permeation studies, were evaluated for their inhibitory activities using croton-oil induced mouse ear edema by topical administration. It was found that the selected transdermal patch containing turpentine oil produced better results as compared with the commercial gel. This was confirmed by histological examination of the inflamed ears treated by the selected patches. A skin irritation study was also performed with turpentine DS patches on rabbit's skin, and no visible reaction was observed. Hence, it can be reasonably concluded that the HPMC patch containing turpentine oil as penetration enhancer was the best choice for manufacturing DS into transdermal matrix type patches to sustain its release characteristics beside its safe properties.




Abdel Naser Singab*, Khaled M. Meselhy**, Asmaa I. Ali***, Amany A. Sleem****

*Pharmacognosy Department, Faculty of Pharmacy, Ain Shams University

**Pharmacognosy Department, Faculty of Pharmacy, Cairo University

***Pharmacognosy Department, Faculty of Pharmacy, Misr International University

****Pharmacology Department, National Research Center, Giza, Egypt.

Certain phytochemical studies on the total oleogumresin of Boswellia carteri Birdwood (Somalia), (Bursereaceae) were carried out and different fractions obtained thereof (gum, oleoresin, volatile oil, and resin). GC/MS (gas chromatography coupled by mass spectrum) was used for qualitative and quantitative analyses of essential oil revealed high percentage of oxygenated constituents (65.29%), while un-oxygenated compounds being (22.63%). The GLC (gas liquid chromatography) analysis of USM (unsaponifiable matter) from petroleum ether extract of oleogumresin revealed 80.96% of hydrocarbons, while sterols being 2.6% of USM. On the other hand fatty acids are composed of saturated acids (47.5%) and unsaturated ones (44.54%). HPLC (high performance liquid chromatography) was used for qualitative and quantitative analyses of gum hydrolysate where glucouronic acid represented the major component (38%). Boswellic acid and two boswellic acid derivatives (3-O-stilbene boswellic acid and acetyl boswellic acid) were isolated from the resin after removal of gum and oil. Major diterpene Incensole was isolated from hydrolyzed resin. Five tested samples prepared from oleogumresin (total aqueous, gum, oleoresin, oil and resin) exhibited significant bioactivities including: analgesic, anti-inflammatory, antipyretic, antioxidant, chronic anti-hyperglycaemic, hepato-protective, antiwormal and antimicrobial activities. In addition a remarkable immunstimulant effect was exhibited by total aqueous extract, as well as gum.



Olaya A.El-Bahrawy*, Reem H. Shaker*, Mai M.H. Selim** and Hala H. Mohamed*

Anatomy* and Histology** Departments, Faculty of Medicine for Girls,

Al-Azhar University, Nasr City, Cairo, Egypt.

Introduction: Heavy metals are widely distributed in the environment, some of them are present in food, water and air. Even in the absence of occupational exposure. Although lead is one of the most dangerous heavy metals, specially toxic to children, it is used in many domestic and cosmetic purposes. Kidneys can remove some of the toxic products of lead metabolism from the blood stream by their excretion in urine. Aim of the work: This work is planned to study the relation between the effect of lead acetate on the kidney structure of infant albino rats at variable duration and forms of exposure. For example indirect lead exposure of offsprings through their pregnant mothers or direct exposure of the offsprings. Materials and methods: 18 offsprings of albino rats were used in the present experiment. The offsprings were divided into 3 main groups. Each group was divided into 2 subgroups according to the age of the offsprings (2 weeks & 1 month).Gr I: The control group. Gr II: Offsprings of exposed pregnant mothers (the lead was given to the pregnant mothers). Gr III: The exposed offsprings group (the lead was given to the offsprings after birth). At the end of the experiment the kidney specimens were collected, processed and subjected to light and electron microscopic studies. Results: Exposure to lead sublethal doses, whether directly to the offsprings or indirectly through exposure of the pregnant mothers, affected the kidney histogenesis and caused certain destructive changes in the renal cortex. Whatever type of lead exposure (direct or indirect) delayed the maturity of the kidney. Conclusion: Lead has harmful effects on the cortical kidney tissue specially during the period of embryogenesis and early fetal life. Thus it is advisable to avoid lead exposure especially during pregnancy and early fetal life. Key wards: lead acetate, kidney histogenesis, offsprings, pregnant female, environmental pollution.



Khalid A.Attia, Hamed M.Abou-Seada, Monir A.Amin and Ragab A.Said.

Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University,

Cairo, Egypt.

Five simple, sensitive, accurate and precise methods are developed for determination of gatifloxacin (GTF) in bulk and in pharmaceutical preparations. The first method depends on charge transfer complexing of GTF with 2,3-Dichloro-5,6-Dicyano-1,4-Benzoquinone (DDQ) to produce purple red colour measured at 587 nm. Beer’s low was obeyed in the range of 10 –80 µg ml-1 with LOD of 0.92 µg ml-1 and LOQ of 3.1 µg ml-1. The second method depends on ion pairing of GTF with eosin-Y dye at pH 1.6 to produce red coloured complex measured at 541 nm. Beer’s low was obeyed in the range of 1 – 9 µgml-1. LOD and LOQ were found to be 0.243 and 0.810 µg ml-1 respectively. The third method depends on the reaction of GTF with 1, 10–orthophenanthroline Fe (III) reagent and the obtained red coloured reaction product measuring at 510 nm. A linear relationship between absorbance and concentration was obtained in the range of 1 – 8 µg ml-1.LOD and LOQ were found to be 0.101 and 0.338 µg ml-1 respectively. The fourth method is an HPLC stability – indicating one where the intact drug GTF, internal standard (ceprofloxacin) and GTF degradation product were separated using a Scharlau nucleosil C18 column (250 mm x 4.6 mm ID x 5µm) with a mobile phase consists of 0.05 M KH2PO4 containing 0.6 % triethylamine adjusted to pH 3 using orthophosphoric acid 85%: acetonitrile: tetrahydrofuran (81 : 18 : 1 by volume) at a flow rate 1 ml min-1 and UV detection at 293 nm. A good linearity was obtained in the range of 0.25 – 5µgml-1.The LOD was 0.055 µg ml-1 and the LOQ was 0.183µgml-1 .The last method is stability – indicating First – Derivative (1D) one for the determination of intact GTF in presence of its degradation product at 300 nm in the range of 2 – 20 µg ml-1 with LOD of 0.036 µg ml-1 and LOQ of 0.12 µg ml-1. The percent recoveries of these methods are 100.19 – 100.77 %. The obtained results were compared with those of the reported method and no significant difference was observed regarding accuracy and precision.

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