Determination of Adulterants in Honey Using HPLC Method

Determination of Adulterants in Honey Using HPLC Method Determination of adulterants such as hydroxymethylfurfural (HMF) in honey using HPLC method 2. Materials and Methods 2.1 Materials Standard of hydroxymethylfurfural was purchased from sigma Aldrich. All the samples and standards were diluted using distilled deionised water. Methanol, sodium hydroxide,diphenyl-1-pikryl, ascorbic acid, 2,2- hydrochloric acid and acetic acid were of analytical reagent grade and purchased from Techno PharmChem, Avonchem and Alpha Chemika Ltd. Most glassware used are stainless steel dish, porcelain crucible, beakers (5, 25, 100 and 250 mL), pipettes (1, 10 and 50 mL), burette (10 and 25 mL), measuring cylinders (10, 100, 250 and 500 mL) and volumetric flasks (10, 100, 200, 1000 mL) were of grade A. All the glasswares were rinsed with distilled deionised water to remove surface contaminants prior to use. Oven, Incubator and furnace used were of make Gallenkamp, Memmert and respectively The absorbance was recorded on a Perlong DNM-9602 Microplate Reader. 2.2 Description of honey sample 13 Honey samples of different brands (Wescobee, Elodie, EL BREZAL, Hosen, Tropic, Lune de miel, Mc Mahon’s, ALSHIFA, Casino, Sunny, Gitto’s, Rodrigues Honey, and Natural Honey) were bought from Intermart Hypermarket, Jumbo Hypermarket and Monoprix Hypermarket. Brief details of different honey analysed, highlighting the manufacturing and expiry date, lot no, country of origin, and ingredients (if specified) are given in table 5. Table 5: Description of honey samples Sample code Sample Brand/ Name Details SP1 (a) Wescobee 100% Pure Honey Manufacturing date: May 2014 Expiry date: May 2017 Lot no:4107A Origin: Australia Ingredient: Pure honey SP9 (b) Elodie Miel Fruità © Manufacturing date: NA Expiry date: 28.08.15 Lot no: 4059ID Origin: France Ingredient: NA SP3 (c) EL BREZAL Eucalyptus Honey Manufacturing date: February 2014 Expiry date: February 2017 Lot no: 2247 Origin: Spain Ingredient: 100% pure natural honey SP8 (d) Hosen Pure Honey Manufacturing date: 25.10.13 Expiry date: 24.10.15 Lot no: NA Origin: China Ingredient: 100% honey SP6 (e) Tropic Lychee Honey Manufacturing date: September 2012 Expiry date: August 2014 Lot no: TP-002 Origin: India Ingredient: Pure honey SP10 (f) Lune de miel Miel de fleur Manufacturing date: NA Expiry date: 01.05.16 Lot no:L28663A Origin: France Ingredient: 100% pure honey SP5 (g) Mc Mahon’s Pure Honey Manufacturing date: NA Expiry date: March 2016 Lot no: B140813 Origin: Australia Ingredient: NA SP7 (h) ALSHIFA Natural Honey Manufacturing date: November 2013 Expiry date: October 2018 Lot no: NA Origin: Saudi Arabia Ingredient: 100% natural SP4 (i) Casino Miel de fleur Manufacturing date: NA Expiry date: 10.03.16 Lot no: 206354 Origin: France Ingredient: NA SP12 (j) Sunny Pure Honey Manufacturing date: NA Expiry date: 24.01.16 Lot no: NA Origin: Mauritius Ingredient: Honey SP11 (k) Gitto’s Special Honey Manufacturing date: 27.06.14 Expiry date: 26.06.15 Lot no: NA Origin: Mauritius Ingredient: Cane sugar syrup 85%, pure honey 15% SP2 (l) Rodrigues Honey Manufacturing date: NA Expiry date: NA Lot no: NA Origin: Rodrigues Ingredient: NA SP13 (m) Natural Honey Manufacturing date: NA Expiry date: NA Lot no: NA Origin: Mauritius Ingredient: NA 2.3 Methods 2.3.1 Sample preparation prior to physicochemical analysis 2.3.1.1 Moisture Moisture was determined according to AOAC method (925.45D) (Appendix I). Stainless steel dish with 25g sand and a glass rod were dried in an oven for 1 hour, cooled in a desiccator then weighed. 1g of homogenised honey sample was added and mixed thoroughly with the sand by means of the rod. The latter was then heated on steam bath for 15 min and dried in an oven for 2 hours at 60 °C, removed, allowed to cool in desiccator and weighed to a constant mass. 2.3.1.2 Ash Ash was determined according to AOAC method (920.181) (Appendix II) such that 5g of homogenized honey samples were added to pre-weighed empty porcelain. The samples were then allowed to dry on a water bath and heated on a hot plate until carbonized. The resulting carbonized samples were place in furnace at 600 °C for 6 hours, removed, allowed to cool in desiccator and weighed. 2.3.1.3 pH pH was measured at 25 °C by preparing a 10% (w/v) solution (dry weight basis) in distilled deionised water by a Delta Ohm HD 8706 pH meter. 2.3.1.4 Total Acidity Free acids, lactones, total acidity and pH were measured using a Mettler Toledo MP 220 pH meter according to the AOAC method 962.19 (Appendix III) as follows: 10g of honey samples were weighed in a 250 mL beaker and dissolved in 75 mL of CO2 free distilled deionised water (obtained by cooling freshly boiled deionised water). The mixtures were stirred using magnetic stirrer and titrated against 0.05M sodium hydroxide at a rate of 5 mL/min until the pH reached 8.50. 10 mL of 0.05M sodium hydroxide was pipetted and back-titrated with 0.05M hydrochloric acid to pH 8.30. A blank titration was also performed using similar procedure. Acidity of honey samples were calculated as follows: 2.3.1.6 Electrical conductivity The electrical conductivity was determined based on a method derived from Apiservices from the ash content of the honey samples according to the equation: 2.3.2 Sample preparation for HMF determination prior to HPLC analysis Aliquots of honey samples were prepared by weighing 1g of honey and were diluted to 10 ml with distilled water, filtered on 0.45 mm filter and injected into an HPLC equipped with a UV detector. The HPLC column was an Agilent, C18, 5 µm, 125 x 4 mm. The HPLC conditions were the following: isocratic mobile phase, 1% of acetic acid and methanol in the ratio (80:20); flow rate, 0.25 ml/min; injection volume, 2  µl, temperature; 30 °C. All the solvents were of HPLC grade. The chromatograms were monitored at 285 nm. All the samples were analysed in triplicates and after every 6 samples, a standard check was analysed. HMF was identified by analysing the peak in honey with a standard HMF, and by comparison of the spectra of the HMF standard with that of one honey samples. The amount of HMF in the honey samples was determined using a calibration curve and by comparing the peak area of the standard and the resulting samples. All honey samples were stored at room temperature (25–30 °C) in a well closed container and the HMF content of each sample was analysed on a monthly basis throughout a period of four months. To determine HMF progress during heat treatment, honey samples were subjected to heat treatment by placing 1g honey sample in a glass container, and heat in a water bath at 40 ºC, 60  ºC, 80 ºC, and 100 ºC for 5 min. The time was calculated when temperature reached the required degree. The honey samples were then cooled rapidly to room temperature (25 °C) and proceed as above to determine the HMF content. 2.3.2.1 Calibration curve for HMF for HPLC analysis A 100 ppm stock solution of HMF standard was prepared by dissolving 0.0101 g of HMF standard in 100 mL of distilled deionised water in a 100 mL volumetric flask. From the 100 ppm stock solution, 10 ppm, 20 ppm, 30 ppm, 40 ppm and 50 ppm standard solution were prepared separately in 10 mL volumetric flasks. The different volumes of the stock solutions which were diluted to 10 mL are given in Table 4. The resulting standards were analysed on a HPLC UV detector at 285 nm and a calibration curve was plotted. Table 6: Volume of 100 ppm stock to prepare different concentration of HMF Concentration/ppm Volume of 100 ppm stock used/mL 10 1.00 20 2.00 30 3.00 40 4.00 50 5.00 A 10 ppm spike sample of HMF was prepared by pipetting 200  µL of 100 ppm stock solution of HMF standard and transferred to the 2 g sample of honey and diluted to 20 mL with distilled deionised water. 2.3.2.2 Limit of detection and quantification The limit of detection and quantification of HMF was calculated according to EPA method SW-846 (Appendix V). LOD is defined to be the minimum level at which the analyte can be detected reliably with signal-to noise 3:1. Different standards of HMF was analysed namely 0.1, 0.05 and 0.04ppm such that detection limit of HMF was thus then established by analysing a 0.05ppm HMF standard solution seven times and the standard deviation of the repeats for the analyse was multiplied by a factor 3.14 based on student t-statistics. The limit of quantification with signal-to noise ratio 10:1 was calculated by multiplying the obtained standard deviation by 10. 2.3.3 Anti-oxidative property using DPPH radical scavenging activity 4.5mg of DPPH (1, 1-diphenyl-2-picrylhydrazyl) was dissolved in 100 mL methanol and wrapped in aluminium foil to prevent light from entering. For the assay, a 96 wells Elisa plate was used. 100 µL of test sample was placed in the first well using micropipette. 50 µL methanol was added to all other wells and serial dilution was done. 50 µL of sample from the first well was pipette and transferred to the second well previously containing 50 µL methanol and the solution was mixed to ensure homogeneity. The 50 µL of the resulting solution was pipette and transferred to the third well and so on. Each well now contained 100 µL of solution after the dilution. 100 µL DPPH (4.5mg/100ml) solution was then added to every well. The solutions were incubated for 30 minutes at 37 °C in an incubator and the absorbances of the resulting solutions were read at 492nm on a Perlong DNM-9602 Microplate Reader. The % scavenging activity of the samples was calculated as follows: Antioxidative property of the samples; firstly with no heating of the honeys and secondly with a heating temperature of 100 °C for five minutes were performed and proceed similarly to that of control ascorbic acid. Note: A yellowish change in colour indicates the presence of ascorbic acid activity. 1311

My Philosophy :: Education Teaching Essays

My Philosophy Education is important because it serves as a foundation for your quality of life. The over all purpose of any education is to teach children the world. It gives you a structure that is always changing as you learn more and more everyday. I want to become a physical education and health teacher because I feel that the education of a healthy lifestyle and active play need to be incorporated into everyone’s lives. I hope to provide students with the knowledge they need to stay healthy and to desire to be that way. What is my philosophy in teaching? Well, I am eclectic and have a few that I fit into. I find mainly that I am a progressivist. I also agree with certain aspects from the philosophical stances idealism, realism, and existentialism. Being a physical education major, I really do not see how I could become a traditional teacher. I do not picture myself taking on much of a democratic role, especially in the gym setting that I would primarily be in. I assume that I will tak e up the role of a facilitating teacher and have student centered activities where we will learn new things together. I plan to give my students the freedom to make their own decisions for the most part, because no one is going to have fun and want to participate much at all if they are not in an enjoyable setting. There will be rules that my students and I will determine together, and these are what will remain for the entire duration of the class as long as they do not take advantage of the rules or myself. In a way, I will give my students the opportunity to lead their class, as long as they do it responsibly and do not get out of hand. I think that my curricular area is important because of the rise of childhood obesity in the United States today. I hope to contribute, in a way, to the decline of obesity in the future. As a teacher I hope to accomplish the impossible, as I am sure that everyone sets out to do in any field. I want students to leave my class feeling like I care about them and that they belong in my class as well. I want to be one of the teachers that the students look up to and feel comfortable and at ease around. My Philosophy :: Education Teaching Essays My Philosophy Education is important because it serves as a foundation for your quality of life. The over all purpose of any education is to teach children the world. It gives you a structure that is always changing as you learn more and more everyday. I want to become a physical education and health teacher because I feel that the education of a healthy lifestyle and active play need to be incorporated into everyone’s lives. I hope to provide students with the knowledge they need to stay healthy and to desire to be that way. What is my philosophy in teaching? Well, I am eclectic and have a few that I fit into. I find mainly that I am a progressivist. I also agree with certain aspects from the philosophical stances idealism, realism, and existentialism. Being a physical education major, I really do not see how I could become a traditional teacher. I do not picture myself taking on much of a democratic role, especially in the gym setting that I would primarily be in. I assume that I will tak e up the role of a facilitating teacher and have student centered activities where we will learn new things together. I plan to give my students the freedom to make their own decisions for the most part, because no one is going to have fun and want to participate much at all if they are not in an enjoyable setting. There will be rules that my students and I will determine together, and these are what will remain for the entire duration of the class as long as they do not take advantage of the rules or myself. In a way, I will give my students the opportunity to lead their class, as long as they do it responsibly and do not get out of hand. I think that my curricular area is important because of the rise of childhood obesity in the United States today. I hope to contribute, in a way, to the decline of obesity in the future. As a teacher I hope to accomplish the impossible, as I am sure that everyone sets out to do in any field. I want students to leave my class feeling like I care about them and that they belong in my class as well. I want to be one of the teachers that the students look up to and feel comfortable and at ease around.

The Associations Anxiety

The Associations between Anxiety and Psychological and Biological Factors Many people believe that anxiety exists in the occasions that give you a lot of stress. Presence of anxiety is mainly due to the environmental elements. In my opinion, anxiety is a result of imbalance of internal states of human being rather that purely caused by environmental factors. The external environment only triggers the disequilibrium of mental status but not all the person will experience anxiety under a same stressful situation. But first, we must understand what anxiety is and how it is formed.In this report, some case studies in General Anxiety Disorder (GAD) help to illustrate my point. First of all, anxiety is a combination of different emotions like nervousness, fear, apprehension, and worrying. It is different from stress. Stress is only an unpleasant state in our body and leads to some biological arousal such as increase in heart beat rate and breathing rate. Stress can be caused by several kin d of emotion like nervous, embarrass or fear which is usually caused by external stimulus. Stress will disappear without the presence of the stimulus. However, for anxiety, formation of tress is more than environmental irritation.Under the same situation, some people will feel anxious but some may not. Different person have different level of stress and anxiety towards same stimulus. Thus, there must be something other than external causes in appearance on anxiety. It may be more related to the cognition of the person which is how the person Judge and value an object or issue as different people have various perceptions and determinations. In understanding which type of person with certain kind of Judgment tends to experience anxiety rather than Just tress, psychological factor including ego and personality can give an explanation to this phenomenon.Other than psychological aspect, some biological factors also lead to creation of anxiety in GAD patients. First of all, anxiety tends to associate with ego. A person with a weaker ego tends to experience stronger anxiety in high frequency. It can be explained by the model proposed by Sigmund Freud. Ego is to please the id which is referred to humans basic instincts, sex and aggressive, and at the same time it also mediate id with superego which is the moral of human beings. Ego helps maintain a healthy and stable psychological state and solve the internal conflicts between id and superego.When the internal conflicts cannot be solved, anxiety will appears which is an unhealthy emotions affecting people's behaviors. There are several factors leading to a weak ego but the main concern is the childhood development of the person which is further illustrated by a Neo-Freudian psychiatrist Harry Stack Sullivan. In Sullivan findings, person with unhealthy growth in childhood block one in building a healthy ego and hence affecting the developing healthy personality. People have unpleasant experience in childhood may result in stronger id or weaker superego.So, ego is not able to balance them and affect the development of healthy personality. Their behaviors may deviate from social norms which make them feel anxious when receiving negative consequences. Also, when people receive condition parental love, they tend to have weaker ego and lower self-esteem. Since their parents only express their cares to the children when they have good performance, they cannot accept their faults and always attribute the reason of failure towards homeless. Once they cannot fulfill their expectations or goals, they will suspect their own abilities and afraid of being blame by others.They overlook the environmental factors and overestimate personal factors. Ego is being destroyed as the central value has been doubted or even denied. This can explain the reason of people having weak ego and unhealthy personality tends to experience intense anxiety in higher frequency and thus proving the association between ego and anxiety . In a recent research done by Assai and her group mates (201 1), it reveals the relation between psychological and behavioral control and self-reported GAD symptoms. It is found that children's behaviors are greatly influenced by parents.Conditional love is actually a kind of parental control that love will be given only following parents' instruction. Under this circumstances, children do not receive chance to develop ego and understand who they are and what they really want to do. They over-rely on their family and thus cannot bearing risk. When they step out into school or society, they have to deal with different challenges individually. Therefore, heir ability to handle problems will be lower and thus experience higher stress level. Some of them may even anxious about things do not have to worry.It shows that excessive parental control which is an unhealthy childhood development leads to the symptoms of over-reaction toward a minor issue. Even though the object or event is sti ll ambiguous and not imposing direct danger to the person, (s)he is daddy interpret it as a threat and (s)he cannot adore anymore. Anxiety arouses and interfere the normal behaviors. In the studies carried by Boston Children's Hospital, children with GAD will anxious in true events, past behaviors, social acceptance, family matters, personal abilities and perceived personal.It has no â€Å"on-off' switch for the worries and they tend to get extreme perceptions. For normal children, they will still have these worries but it usually appears in the present of certain stimulus or occasions which trigger your thought. Stress on them is specified on targeted issue and will disappear after all. However, children with GAD are not easy to relax and concentrate since the stimulus for their worries refer to lower self-esteem and weaker ego which permanently exists before any changing to improve it. In the world, most of people have healthy and happy childhood period.Nevertheless, some of them still get anxious frequently. The appearance of anxiety seems not only associated with ego. In biological field, scientist studied the heredity of which characteristics, both superior and inferior, can be carried down to next generation. Personality can also be inherited by parents. People with type A personality is ambitious, rigidly, sensitive, impatient. They cannot easily accept failure. A little setback will trigger their nervousness and take precautions in order to turn the thing back to correct track. In there words, they have low adaptability and over-react to changes.They will have relatively higher stress level than normal person. Type A personality can be genetically inhered by last generation but also be shaped by environment. People staying at the high tension state over a long period of time, they will more likely feel anxious. For example, people work in the company which required high accuracy and stable performance with long working hours. They will experience extr emely high level of stress for long time. They afraid of the consequences of making a mistake and thus need to double check and make sure everything is competed and correct before submitting to boss.Those people may have chance to become perfectionism which they will force themselves act perfectly at any moment. They may be rigid that they cannot handle problem in a more flexible way. They will be impatient and try to finish all the cases as fast as they can. They are not able to relax and anxious of something that does not need to worry about at that time. Moreover, the deficiency of brain may also leads to anxiety. Some of GAD patients are diagnosed that part of the brain component or levels of neurotransmitters are abnormal. In psychological studies, neurotransmitters are important in regulating humans behaviors and mental health.Especially for GAB, serotonin and endorphins, they are relatively significant in maintaining emotions. GAB is an inhibitory neurotransmitter which contr ibutes to canceling the effects of the excitatory SSP. Decrease in level of GAB leads to anxiety as the person will always has high activity level in brain transmission and keep him at a relatively high tension state. Serotonin involves in regulation of sleep and wakefulness, eating and aggression. Abnormal amount of serotonin leads to difficulty in getting sufficient rest and become aggressive. Endorphins help to relief pain and some pleasurable emotions.Lacking of endorphins make the person hard to neither enter into positive emotions nor eliminate the negative moods. The person will become irritable and sensitive and thus not able to relax. A GAD patient, James, whose brain was not functioning well and having imbalance level of various neurotransmitters, turned him into an anxious person. He was easily distracted by minor thing and often nervous. He could not sleep well and complained of headache. It serious disturbed his cognition and he could not have clear mind to make Judgmen t. He might than always has bad performance which brings him stress again.Other than neurotransmitter, defective in brain also cause the instability of emotion. Other than abnormal level of neurotransmitters, James also suffered from malfunction of temporal lobe located in cerebrum which processes and coordinates stimuli received for learning, thinking, remembering and conscious awareness. Temporal lobe is responsible in process of various sounds and sights stimuli and managing the long term memory storage. As a result, he had difficulties in memorizing instructions and recognizing the work. He always forgot simple procedures.By comparing to normal person that can finish targets, James tends to have weak self-esteem and thus give a lot of pressure and blame himself of the failure. Besides, malfunction of some emotion centers such as magical also lead to GAD. Researchers have found that some GAD patients have larger volume of magical. Magical plays a central role in the fear response and aggression. Also, it promotes the fight or flight response which they person will either attack or escape hen facing dangerous. The enlarged magical is easier be triggered to alter people and hence creating fear and aggression.It can explain why the patient will feel anxious to stimuli which do not carry any threat. Brain activity is influenced by the magical so that they always get nervous. Therefore, existence of anxiety is affected by the brain structures. All people experience stress but people with psychological or biological problems tends to develop anxiety over same level of pressure. People act differently towards under the same environment the aforementioned evidence and studies suggest a utter way to understand anxiety that psychological and biological factors may have greater association with anxiety.

The Media Responsible For Females Essay - 1679 Words

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Females, especially young girls are developing disorded eating or an eating disorder as a result of the media promoting the super slender body, and with advertisements encouraging consumers to purchase high in fat foods at the same time, is leading to more individuals becoming obese. The space between an ordinary female and an ‘ideal’ female is widening, to which causes an increase in anxiety and to help overcome the feeling of anxiety, a decrease in body weight seems to be the answer. A diet isShow MoreRelatedGender Inequality According to Functionalist and Marxist Feminist Perspective900 Words   |  4 Pagessociety; the film Missrepresentation, by Jennifer Newsom reveals the lack of female presence in lead or authoritative roles in media, in comparison to men (Newsom, 2011). The film Missrepresentation, by Jennifer Newsom, is about the underrepresentation of women by the media (2011). 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