Tuesday, January 28, 2020

RP-HPLC Method for Estimation of Ferrous Ascorbate

RP-HPLC Method for Estimation of Ferrous Ascorbate Development and validation of RP-HPLC method for estimation of ferrous ascorbate and folic acid in bulk and in pharmaceutical formulation ABSTRACT A simple, sensitive, precise and reproducible RP-HPLC method has been developed and validated for estimating the contents of ferrous ascorbate and folic acid combination in bulk and pharmaceutical formulation. The analysis was performed with Gemini-NX-C18 (250Ãâ€"4.60 mm, 5 µm) column using mobile phase phosphate buffer (pH 7): methanol (98:2, v/v) for ferrous ascorbate whereas phosphate buffer: methanol (88:12, v/v) for folic acid. Mobile phase was delivered at flow rate of 1.0 ml/min for ferrous ascorbate and folic acid. The detection wavelength was 264 nm for ferrous ascorbate and 280 nm for folic acid. The retention time of ferrous ascorbate and folic acid was found to be 2.7 and 6.8 min, respectively. The linearity was obeyed in the concentration range of 10-50  µg/mL for ferrous ascorbate (r2 = 0.999) and 10-50  µg/mL for folic acid (r2= 0.999). Tablet content of ferrous ascorbate and folic acid was found 100.44 % and 98.69 %, respectively. INTRODUCTION Iron deficiency and sickle cell is the most common cause of anemia worldwide, accounting for about half of all anemia cases. Prevalence of anemia in India is high because of low dietary intake, poor availability of iron and chronic blood loss due to hookworm infestation and malaria. An exogenous source of folic acid is necessary for the maintenance of normal erythropoiesis. Study was carried out to assess efficacy, safety and tolerability of combination of ferrous ascorbate and folic acid (Phosfomin-XT) in patients with iron deficiency anemia, the report sho wn the increase in bioavailability of ferrous in presence of ascorbate salt. ferrous ascorbate (Fig. 1.A) is newly introduced salt of ferrous having more bioavailability than other salts of ferrous ascorbate [1]. But till date no more literature available for estimation of ferrous ascorbate. The use of therapeutic multivitamins are indicated in cases of deficiency in pathological conditions in which the nutritional requirements a re greatly increased or in conditions in which absorption, utilization, or excretion of vitamins are abnormal [2]. Folic acid (Fig. 1.B) deficiency is the result of megaloblastic anemia [3]. Ion pair chromatographic method was reported for simultaneous determination of seven water soluble vitamins nicotinamide, thiamine, riboflavin, pyridoxine, pyridoxal, pyridoxamine, cyanocobalamine and folic acid [2]. HPLC method was developed for estimation of folic acid [2-4]. RP-HPLC method for estimation of folic acid was also reported in Indian pharmacopoeia (2010) [5]. But till date, no report is available for estimation of ferrous ascorbate in combination with folic acid in bulk and pharmaceutical formulation. An attempt has made to develop and validate a new, rapid and sensitive method for estimating the content of ferrous ascorbate and folic acid. Experimental Conditions Instrumentation The HPLC chromatographic system Agilent Technologies 1200 series, assembled with a G1315D quaternary pump, a G1315D diode array detector, a rheodyne injector fitted with a 20 ÃŽ ¼L loop and EZ Chrome Elite software. Reagents and chemicals Ferrous ascorbate and folic acid were obtained as a gift samples from Curex Pharmaceutical Pvt. Ltd., Jalgaon (MS) and S. kant Healthcare Ltd, Wapi (gujrat). HPLC grade acetonitrile, and methanol was purchased from Rankem Ltd., India. Double distilled water was generated in house. IPHARED tablets (ferrous ascorbate equivalent to elemental iron and folic acid in ratio of 100:1.5 mg) were purchased from a local market. Standard stock solution of ferrous ascorbate (100  µg/mL) An accurately weighed quantity of about 10 mg ferrous ascorbate was dissolved in 100.0 mL of phosphate buffer (pH 7). Working standard solution of ferrous ascorbate Working standard solution of ferrous ascorbate in the concentration range of 10–50 ÃŽ ¼g/mL was prepared by diluting an aliquot portion of about 1– 5 mL ferrous ascorbate stock solution (100 ÃŽ ¼g/mL) to 10 mL with mobile phase. Standard stock solution of folic acid (100  µg/mL) An accurately weighed quantity of about 10 mg folic acid was mixed with 0.5 mL 2.86% solution of sodium carbonate and diluted it upto 100 ml with mobile phase [5]. Working standard solution of folic acid Similarly working standard solution of folic acid in the concentration range of 10–50 ÃŽ ¼g/mL was prepared by diluting an aliquot portion of about 1–5 mL folic acid stock solution (100 ÃŽ ¼g/mL) to 10 mL with mobile phase. Sample preparation for Tablet analysis An accurately weight 20 tablets were crushed to obtained fine powder. Sample preparation of ferrous ascorbate A quantity of tablet powder equivalent to 10 mg of ferrous ascorbate (11.31 mg) was dissolved in 100 mL phosphate buffer pH 7, sonicated for 30 minutes. The solution was filtered. The aliquot portion of filtrate (2.0 mL) was diluted to 10.0 mL with mobile phase. Sample preparation of folic acid A quantity of tablet powder equivalent to 5 mg of folic acid (2.72 g) was mixed with 0.5 mL of 2.86% of sodium carbonate solution and then dissolved in 100 mL mobile phase. The mixture was sonicated for 30 minutes. The solution was filtered. An aliquot portion of filtrate (4.0 mL) was further diluted to 10.0 mL with mobile phase. Chromatographic condition ferrous ascorbate and folic acid Chromatographic condition ferrous ascorbate The optimized chromatographic conditions for ferrous ascorbate estimation were Gemini-NX- C18 (250Ãâ€" 4.60, 5 µm) as a stationary phase, mixed phosphate buffer (pH 7): methanol (98:2, v/v) as mobile phase, flow rate 1 mL/min with detection wavelength 264 nm. The chromatogram for ferrous ascorbate is as shown in (Fig. 2). The optimized chromatographic conditions for folic acid estimation were Gemini-NX- C18 (250Ãâ€" 4.60, 5 µm) as a stationary phase, phosphate buffer: methanol (88:12, v/v) as mobile phase, flow rate 1 mL/min with detection wavelength 280 nm. The chromatogram for ferrous ascorbate is as shown in (Fig. 3). Results and discussion Ferrous ascorbate and folic acid combination was used in management of anemia [6]. In literature various HPLC methods for estimation of folic acid was reported [7-9]. But no method was reported for the estimation of ferrous ascorbate and simultaneous estimation of ferrous ascorbate and folic acid combination. A numbers of trials were carried out for simultaneous estimation of ferrous ascorbate and folic acid. Initially, therefore attempt was done for simultaneous estimation of both drugs by reversed-phase, HPLC separation was tried using phosphate buffer pH 3 and methanol (98:2) as the mobile phase, in which ferrous ascorbate eluted at 2.4 min which is nearer to dead time of column and folic acid was not eluted, the resolution and numbers of theroetical plates was also poor. The organic content of mobile phase was also investigated in different ratio to optimize the separation of ferrous ascorbate and folic acid simultaneously. But no mobile phase was found suitable for analysis. The refore, attempt was made to estimate ferrous ascorbate and folic acid in bulk and in pharmaceutical formulation separately using different chromatographic condition which was optimized separately for ferrous ascorbate and folic acid (Table I). Therefore attempt was made to estimate ferrous ascorbate and folic acid by using two different mobile phases and sample preparation method. The mobile phase phosphate buffer pH 7- Methanol (98:2) was selected for ferrous ascorbate estimation. The buffer-methanol (88:12, v/v) was selected for folic acid estimation as per Indian pharmacopoeia [5]. The constant flow rate 1mL/min was maintained for both analysis. Detection wavelength 264 nm for ferrous ascorbate and 280 nm were selected for folic acid. Retention time was found to be 2.7 min for ferrous ascorbate and 6.8 min for and folic acid. VALIDATION PARAMETERS [10, 11]. System suitability System suitability parameters, such as number of theoretical plates, HETP, and peak tailing, were determined. The results obtained are shown in Table I the number of theoretical plates for ferrous ascorbate and folic acid was found 3219 and 2535 respectively with separate mobile phase. Linearity and range Ferrous ascorbate and folic acid follows a linearity of response between 10–50  µg/mL. Linearity study was performed separately by analyzing five different concentrations of ferrous ascorbate and folic acid. The Solution were diluted appropriately with mobile phase to obtain concentrations in the range of 10-50 ÃŽ ¼g/ml. All measurements were repeated three times for each concentration. Y = 23397x + 28729 is the equation for ferrous ascorbate with R2 value 0.999 Y = 188287x +150052 is the equation for folic acid with R2 value 0.999 Precision The repeatability was studied by multiple injections of a homogenous sample of 20  µg/mL of ferrous ascorbate and folic acid separately. The results of repeatability study of ferrous ascorbate and folic acid are as follows. The result of intra-day precision study, % RSD for ferrous ascorbate was found 1.31 and for folic acid was found to be 1.61. The result of Inter-day precision study, the % RSD for ferrous ascorbate was found 1.60 and for folic acid was found to be 1.54 respectively. The results of the precision study indicate that the method is reliable and reproducible, with a relative standard deviation less than 2.0%. Accuracy Recovery study was performed to validate the accuracy of the developed method. Recovery study was performed at 80%, 100% and 120 % level. The known amount of standard ferrous ascorbate was added to preanalyzed sample and subjected for HPLC analysis. Procedure for ferrous ascorbate Sample solution A quantity of tablet powder equivalent to 10 mg ferrous ascorbate (11.35 mg) was dissolved in 100 mL phosphate buffer, sonicated for 30 minutes. The solution was filtered. Standard stock solution of ferrous ascorbate (100  µg/mL) An accurately weighed quantity of about 10 mg ferrous ascorbate was dissolved in 100.0 mL of phosphate buffer pH 7. Working solution of ferrous ascorbate Three sets containing 10.0 ml volumetric flask were taken. To each set, 2 ml of sample solution was added. To the first set (80%) of three volumetric flasks, a quantity of about 1.6 mL of standard stock solution of ferrous ascorbate was added. To the second set (100%) of three volumetric flasks, a quantity of about 2.0 mL of standard stock solution ferrous ascorbate was added. To the third set (120%) of three volumetric flasks, a quantity of about 2.4 mL of standard stock solution of ferrous ascorbate was added. The volume was made up to the mark with mobile phase in all flasks. The sample and standard solution were analyzed as per optimized chromatographic conditions for ferrous ascorbate. Procedure for folic acid Sample solution of folic acid A quantity of tablet powder equivalent to 10 mg folic acid (5360 mg) was dissolved in 1mL of 2.86% of solution of ferrous sodium carbonate and volume was made with 100 mL buffer, sonicated for 30 minutes. The solution was filtered through whatman filter paper No.41. Standard stock solution of folic acid (100  µg/mL) Weighed accurately about 10 mg folic acid mixed with 0.5 ml 2.86% solution of sodium carbonate solution and diluted with 100 ml of mobile phase. Working solution Three sets containing 10.0 ml volumetric flask were taken. To each set, 2 ml of sample solution was added. To the first set (80%) of three volumetric flasks, a quantity of about 1.6 mL of standard stock solution of folic acid was added. To the second set (100 %) of three volumetric flasks, a quantity of about 2.0 mL of standard stock solution folic acid was added. To the third set (120 %) of three volumetric flasks, a quantity of about 2.4 mL of standard stock solution of folic acid was added. Then, the volume was made up to the mark with mobile phase in all flasks. The sample and standard solution were analyzed as per optimized chromatographic conditions given for folic acid. These results of accuracy are as summarized in Table no: II Determination of the limit of detection and quantitation (LOD and LOQ) The limit of detection and limit of quantitation is determined by using following formulae and result is given below. , Where SD = Standard deviation of response, S = Slope of linearity curve equation The result of LOD (ÃŽ ¼g/ml) of ferrous ascorbate and folic acid was found to be 0.259 and 0.213 respectively. The result of LOQ (ÃŽ ¼g/ml) of ferrous ascorbate and folic acid was found to be 0.784 and 0.645 respectively. Robustness As per ICH norms, small but deliberate variations, by altering the flow rate of the mobile phase, were made to check the method’s capacity to remain unaffected (method stability). The change was made in flow rate by altering flow rate 0.9 mL /min and 1.1 mL/min Results of analysis are summarized in Table no: III Tablet analysis Contents of ferrous ascorbate and folic acid found in the tablets by the proposed method are shown in Table VI the low RSD values indicate that the method is precise and accurate. Specificity study Specificity study was conducted for ferrous ascorbate and folic acid in presence of excipients present in tablet. Preparation of physical mixture for specificity study The average weight of 20 tablets was found to be 804 mg. Therefore, as per labelled claim, tablet formulation excipients in tablet were 75 mg. Mixture of drug and excipients {728  µg/mL ferrous ascorbate, 1.5  µg/mL folic acid and 75  µg/mL excipients in equal parts (like lactose monohydrate, microcrystalline cellulose, magnesium stearate, starch pregelatinised etc.)} was prepared. The solution was analyzed by optimized chromatographic conditions given for ferrous ascorbate and folic acid. The peak purity value of ferrous ascorbate and folic acid was found to be more than the input threshold value. It shows that the peak of ferrous ascorbate and folic acid was free from any impurity or co-elution in presence excipients present in tablet. The VU-spectrum of ferrous ascorbate and folic acid is as shown in Fig. 4 and Fig. 5 Also the peak purity spectrum of ferrous ascorbate and folic acid was shown in Fig. 6 and Fig. 7 Conclusion RP-HPLC method for estimation ferrous ascorbate and folic acid in bulk and in pharmaceutical formulation was successfully developed as per ICH guidelines. The method was found accurate, precise and sensetive hence it can be used for routine analysis by any analyst.

Sunday, January 19, 2020

A Winless Battle :: essays papers

A Winless Battle Sometimes in life, there are times when you are faced with a struggle and often times, no matter how hard you try to get yourself out of this struggle, you just can’t and you end up feeling as though you have lost the battle. In â€Å"A Streetcar Named Desire,† by Tennessee Williams, there are four characters that really display this situation. Blanche, Stanley, Stella, and Mitch are the four characters that display this situation. In Tennessee Williams play, â€Å"A Streetcar Named Desire,† there are no victors, only victims. In this play, Blanche is the older sister of Stella and she comes to Elysian Fields to live with her sister because she has nowhere else to live because she was asked to leave the last town that she was living in because of the life that she was choosing to live there. Blanche also lost her family’s estate, Belle Reve. Therefore, she had to go to the last place that she could think of and that was the house of her sister, Stella. From the start of the play, it is imminent that Blanche is the victim of her own life, and not a victor. She first lost her husband when she was a very young girl because of the fact that she found out that he was gay. She was in love with this man and when she found out, she was just heartbroken. However, she didn’t want to tell him that she knew about his secret so she kept it a secret for a while. But then, one night while they were dancing, she all of sudden just came out and told him what she knew. He was so embarrassed by this tha t he ran out of the room and shot and killed himself. That is one of the battles that she was the victim of. Then when she loses Belle Reve and is forced to live in a town called Laurel, she chooses to live a life that she doesn’t want to. She forces herself into prostitution because she has no other way of making money to help support herself. During this time, she is living in a hotel called the Flamingo and many men are coming up to her room every night. This disturbs the peace within the hotel and when the hotel and the town of Laurel figure out what she is doing in the hotel, they tell her to leave the hotel and also the town because they don’t want anyone like her to be there.

Saturday, January 11, 2020

Conflict Analysis Essay

‘When conflict arises, it is impossible not to take a side. Everyone has a role’ Expository Essay: One of the great myths of conflict is that is can always be contained. Whether it is encountered in the personal or political sphere, conflict will often spread beyond the main combatants. History demonstrates that wars escalate in a world where connections between individuals and states are so complex and it is so difficult to come to a definite conclusion. An argument between friends will generally force those on the perimeter to choose sides. Conflict by its very nature creates polarities. Even when conflict is internal, it rarely remains so. When a dispute begins, the world is viewed in black and white terms with no middle ground. Attempts to reconcile differences are often misinterpreted as tacit support for one side or the other. Conflict is defined by incompatible positions that force individuals to take sides. War constructs an environment in which on a political level, everyone is on a side. However, in another sense, everyone is doing what they can or must to survive in their particular situation. Throughout their time as prisoners of war, the women in the camp always disregard the Japanese as their enemy. For some, this is because of nationalistic allegiances, but for other it is because of the way they are treated. Their battle is not against Japan, but against humiliation and the use of brutality to degrade and harm others. The Japanese impose many acts of humiliation upon the women in the camp, such as forcing them to bow and scramble for food, to haul water for Japanese baths, to rise upon order and to assert the supremacy of the Japanese Empire over the British. This humiliation is explained as a reversal of the humiliation to which the Japanese felt the Europeans were subjected colonised Asian people. Hence, why divisions in individuals are experienced during war and consequently force oneself to choose a side. Conflict can rarely be contained and it inevitably spreads beyond the original dispute. Class prejudices often causes disputes between individuals  demonstrating how support and unity is essential in times of conflict. Sometimes the women in the camp are surprised to discover themselves degrading attitudes that they despise in their captors. Prejudices based on class and race must be confronted if conflict is not to destroy all hope and dignity. However, the women do not last long living under the pressures of camp life and soon they are all equal in scrambling for food and bowing to the Japanese flag. Cultural and educational backgrounds have a more profound impact on the relationships formed within the group and arguments raised between individuals. When a racially motivated fight breaks out over a lost bar of soap, it threatens to undermine the unity and strength of the women. This demonstrates the immense importance to disregard social class for these women. The middle ground can be the most dangerous place in a conflict because both sides may regard this position with suspicion. Margaret (Daisy Drummond) frequently seeks to mediate between the Japanese and their victims. Her bravery and compassion and her unshakeable belief in the basic goodness of individuals, puts her life at great risk. Her bravery is evident when she marches out with water for Susan who is weakening from her torture. This action may have led to similar punishment for herself. Sometimes conflict is like a bushfire it spreads and spreads until everyone is forced to take some kind of action. It does not respect those who wish to stay out of its way, anymore than it does those who try to encounter it head on. Arguments, disputes and battles of all sorts are an undeniable part of the fabric of human existence. The goal is always to look for a way to end the fighting and heal the wounds. It is thus crucial not to underestimate the momentum that develops so quickly when conflict arises. Persuasive Essay: We sang wonderfully today. The concert was a huge success! Even Captain Tanaka clapped at the end. This would have been unthinkable a few months ago. Maybe the Japs are beginning to think we are humans after all. Well I don’t think I could ever think of them that way. Adrienne and Daisy were talking about that after the concert. The interpreter asked them to perform a traditional Japanese song. Daisy looked like she was about to agree, but Adrienne said â€Å"NO!† straight away. Adrienne admitted she hates the Japs, but Daisy said the worse they acted the most sorry she felt for them. I’ve been thinking about that a lot. It seems Daisy doesn’t see things like most of us do. She doesn’t see us as innocent and the Japs as evil. She just doesn’t take sides like that. It is easer to wonder if Daisy is like this because she is just too soft-a pushover who will help anyone-maybe just to be liked. I knew some girls like that back at school, they’d get bullied all the time but still hang around with the bullies. But that’s not Daisy. She is the bravest of us all. She marched straight out in front of the soldiers with water for Susan that horrible day and she bargained for Susan’s release too. But that’s it, she wanted to help Susan, but she never wanted to hurt the Japs. For her it isn’t us against them, she feels compassion for everyone. That’s amazing but I don’t know anyone else like Daisy. Most of us are like Adrienne. We have to take sides because the war puts us on a side. I don’t understand how anyone could possibly feel compassion towards the Japs; they have put us through extraordinary challenges, where I personally have had to test my morals. But I have to put aside my morals for the time being, if I want to survive in this corrupt environment. I often relate my thoughts back to Daisy’s, she is so compassionate and highly respected though the camp. Does this mean she accepts the behaviours of the Japs, humiliating us? Daisy was in the missionaries, maybe she feels a sense of understanding towards the Japs? ‘When conflict arises, it is impossible not to take a side. Everyone has a role’ Expository Essay: One of the great myths of conflict is that is can always be contained. Whether it is encountered in the personal or political sphere, conflict will often spread beyond the main combatants. History demonstrates that wars escalate in a world where connections between individuals and states are so complex and it is so difficult to come to a definite conclusion. An argument between friends will generally force those on the perimeter to choose sides. Conflict by its very nature creates polarities. Even when conflict is internal, it rarely remains so. When a dispute begins, the world is viewed in black and white terms with no middle ground. Attempts to reconcile differences are often misinterpreted as tacit support for one side or the other. Conflict is defined by incompatible positions that force individuals to take sides. War constructs an environment in which on a political level, everyone is on a side. However, in another sense, everyone is doing what they can or must to survive in their particular situation. Throughout their time as prisoners of war, the women in the camp always disregard the Japanese as their enemy. For some, this is because of nationalistic allegiances, but for other it is because of the way they are treated. Their battle is not against Japan, but against humiliation and the use of brutality to degrade and harm others. The Japanese impose many acts of humiliation upon the women in the camp, such as forcing them to bow and scramble for food, to haul water for Japanese baths, to rise upon order and to assert the supremacy of the Japanese Empire over the British. This humiliation is explained as a reversal of the humiliation to which the Japanese felt the Europeans were subjected colonised Asian people. Hence, why divisions in individuals are experienced during war and consequently force oneself to choose a side. – Conflict can rarely be contained and it inevitably spreads beyond the original dispute. Class prejudices often causes disputes between individuals demonstrating how support and unity is essential in times of conflict.  Sometimes the women in the camp are surprised to discover themselves degrading attitudes that they despise in their captors. Prejudices based on class and race must be confronted if conflict is not to destroy all hope and dignity. However, the women do not last long living under the pressures of camp life and soon they are all equal in scrambling for food and bowing to the Japanese flag. Cultural and educational backgrounds have a more profound impact on the relationships formed within the group and arguments raised between individuals. When a racially motivated fight breaks out over a lost bar of soap, it threatens to undermine the unity and strength of the women. This demonstrates the immense importance to disregard social class for these women. -indigenous Australians. The middle ground can be the most dangerous place in a conflict because both sides may regard this position with suspicion. Margaret (Daisy Drummond) frequently seeks to mediate between the Japanese and their victims. Her bravery and compassion and her unshakeable belief in the basic goodness of individuals, puts her life at great risk. Her bravery is evident when she marches out with water for Susan who is weakening from her torture. This action may have led to similar punishment for herself. Daisy demonstrates that in times of conflict we can have courage to not comprise our convictions even in the direst circumstances. Her strength comes from her authentic faith. When the observation is made following an incident of nauseating brutality that she doesn’t hate the oppressors, Margaret responds, â€Å"I just can’t bring myself to hate people. The worse they behave, the more sorry I feel for them.† This powerful character illustrates that in times of serve conflict our morals do not have to be compromised if we have the strength to do so. Sometimes conflict is like a bushfire it spreads and spreads until everyone is forced to take some kind of action. It does not respect those who wish to stay out of its way, anymore than it does those who try to encounter it head on. Arguments, disputes and battles of all sorts are an undeniable part of the fabric of human existence. The goal is always to look for a way to end the fighting and heal the wounds. It is thus crucial not to underestimate the momentum that develops so quickly when conflict arises.

Friday, January 3, 2020

Site Admin s Code Of Ethics - 811 Words

Site admin s code of ethics seems to change depending upon the situation and/or person(s) involved. Site admin makes inappropriate personal comments to students and staff, and has a history of openly criticizing others, including students and her own superiors. On the part of parents, I have never heard a kind word spoken about site admin in the many years with the program. When students take the Title 1 survey, they must go to the library and are told by site admin to say nice things, and they do, but about the teachers as shown by last year s results. The parent surveys are sent home on paper and turned in directly to site admin. Morevier, site admin has a policy that office personal are not allowed to talk with other staff members regarding what transpires in the office. Site admin would rather meet with teachers individually than as a group, and has different expectations for different teachers depending upon whether you happen to be favored or not, which changes from day to day, month to month and year to year. Moral is very low and frustration very high among most of the teachers because they do not feel respected, supported, understood or valued. Instead, the majority feel micro-managed, talked down to and regularly picked on. Site admin often engages individual teachers to manage other staff members. 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