Caffeine , Analysis of caffeine

Introduce, properties, comparative analysis

1. Comparative analysis OF CAFFEINE PRESENTED BY GUIDED BY MR. SHUBHAM GAJANAN WAGH PROF. DR.K.B.GABHANE VIDYA BHARATI COLLEGE OF PHARMACY, AMRAVATI-444602 2019-20

2. INDEX Sr. No. Contents Page No. 1 Introduction 3 2 Structure & Properties of Caffeine 4 3 Structural-Activity Relationship 5 4 Mechanism of Action 6 5 Metabolism of Caffeine 7-8 6 Advantages 9 7 Disadvantages 10 8 Identification 11 9 Case studies 12 -15 10 Conclusion 16 11 Reference 17 2 Sr. No. Contents Page No. 1 Introduction 3 2 Structure & Properties of Caffeine 4 3 Structural-Activity Relationship 5 4 Mechanism of Action 6 5 Metabolism of Caffeine 7-8 6 Advantages 9 7 Disadvantages 10 8 Identification 11 9 Case studies 12 -15 10 Conclusion 16 11 Reference 17

3. Introduction Caffeine is a pharmacologically active substance and depending on the dose, can be a mild central nervous system stimulant. It is the world’s most consumed psychoactive drug which is legal and unregulated in most parts of the world. Caffeine is a bitter, white crystalline power and p urine derivative which is a m ethylxanthine alkaloid. 3

4. Structure & PROPERTIES OF CAFFEINE Sr.No. Properties Values/ Appearance 1 Molecular weight 194.19 g/mol. 2 Density 1.2gm/cm 3 3 Melting Point 237 ° C 4 pKa value 10.4 5 Solubility in water Slightly soluble 6 Boiling point 178 °C 7 Appearance White Needles/ Powder IUPAC Name : 1,3,7-Trimethylpurine-2,6 - dione Sy nonyms : Guaranine Methyltheobromine 1,3,7-TrimethylxanthineTheine Molecular Formula : C 8 H 10 N 4 O 2 Structure of caffeine 4

5. Structural-Activity relationship Effects of the structural a ctivity relationship of the xanthenes molecule on adenosine antagonism and systematic effects: Substitution in position 1 is necessary for high affinity and selectivity towards adenosine receptor sites Substitution in position 3 increases bronchodilator effect Substitution in position 7 decreases both adenosine receptor antagonism and bronchodilator potency Substitution in position 9 results in decreased adenosine receptor affinity Substitution in position 8 increases adenosine antagonism and selectivity towards A 1  receptors  X anthines molecule s 5

6. Mechanism of action Adenosine is a central nervous system neuromodulator that has a specific receptors. When adenosine is bind to its receptors neural activity slow down a nd you feel sleepy. Adenosine thus facilitates sleep and dilates the blood vessels. Caffeine act as a adenosine-receptor antagonist. This means that binds to same receptors but without reducing neural activity. Fewer receptors are thus available to the natural braking action of adenosine and neural activity therefore speed's up. Caffeine also stimulate the pituitary gland to secrete the hormones of adrenal gland to produce more adrenaline so it increase your attention level and gives your entire system an extra burst of energy Similarity in double ring structure 6

7. Metabolism Biosynthesis B io synthesis of caffeine is as shown in Fig . Xanthosine is the precursor for caffeine biosynthesis I n the presence of 7-methylxanthosine synthase and S-adenosyl-L - methionine enzymes 7-methylxanthosine is produce from xanthosine. I n the presence of methylxanthosine nucleotidase enzyme gives 7-methylxanthine from 7-methylxanthosine . Later 7-methyl xanthine gives first gives Theobromine and then caffeine in presence of enzymes caffeine synthase and S- adenosyl- l -methionine Paraxanthine is an intermediate synthesized during caffeine biosynthesis which also leads to the synthesis of caffeine. Biosynthesis of Caffeine 7

8. Catabolism The catabolism process is initiated by the degradation of caffeine to xanthine as shown in Fi g . Intermediates such as 3-methyl x anthine, theophylline are formed . B efore the formation of xanthine ,then it is degraded into uric acid , allantoin , allantoate and finally to ammonia and carbondioxide . Caffeine is primarily metabolised in the liver by cytochrome P450 enzymes, which are responsible for more than 90% of caffeine clearance . Catabolism of Caffeine 8

9. Advantages When caffeine is consumed in lower doses (20-200) mg it has the following advantages on the body . Caffeine inhibits adenosine which has an inhibitory effect on neurotransmitters. This increases the activity dopamine . It causes relief from tension and headache. The chances of suffering from depression also reduce . The risk of hepatocellular and endometrial cancer also reduces. It acts as a thermogenic agent by inhibiting the degradation of intracellular cAMP caused by phosphodiesterase .This helps in weight control C affeine has an anticariogenic effect that helps in preventing dental caries which is caused by the production of acid by various types of bacteria such as Streptococcus mutants, Actinomycetes. C affeine helps in reducing blood cholesterol levels and fat absorption by the body and hence reduces obesity. This helps in reducing the risk of a atherosclerosis. It acts as an antioxidant due to the presence of various polyphenols, it helps in preventing oxidative stress in vivo. It also increases the activity of leucocytes . 9

10. Disadvantages When caffeine is consumed in excess, it brings health hazards. They are as follows : Excess caffeine reduces the time a person is able to sleep. It is because of this phenomenon that people suffering from insomnia . According to earlier studies , adenosine causes vasodilatation. Presence of caffeine stimulates the release of nor epinephrine from the adrenal glands which causes vasoconstriction in turn causes an increase in heart rate and blood pressure . Studies have shown that caffeine increases the stiffness of arteries causing a decrease in elasticity of arteries. A high dose of caffeine (>1000 mg) causes caffeine addiction. Anxiety is the main symptom of caffeine addiction, there is an increase in the risk of bladder, the condition of patients suffering from glaucoma is worsened by the increase in pressure within the eye. There is an increase in the risk of osteoporosis due to decreased calcium absorption by bones and increased calcium secretion in urine. The overdose of caffeine also causes premenstrual symptoms in women and there is an increase in the secretion of insulin by the β cells of the pancreas, thus increasing the risk of diabetes. 10

11. IDENTIFICATION Murexide Test The  murexide test  is an analytical technique to identify the presence of  caffeine  and other  purine derivatives  in a sample. These compounds do not respond to the common alkaloids detecting tests such as  Dragendorff’s reagents. In this test the alkaloids are mixed with a tiny amount of  potassium chlorate  and a drop of  hydrochloric acid. The sample is then evaporated to dryness and the resulting residue is exposed to  ammonia  vapour. Purine alkaloids produce a pink colour in this test. Murexide (ammonium purpurate) with a purple colour is also produced in this test . 11

12. Case studies Fourier transform infrared spectroscopy has proved to be a good technique for quantifying a wide range of materials especially pharmaceutical products when it is used in combination with Attenuated Total Reflectance Accessory. TGA/DTA a combined technique in thermal analysis measures the physical changes and chemical changes that occur in characterizing the samples to show thermal stability and material purity . Apparatus FTIR Nicolet 6700 equipped with Attenuated Total Reflectance accessory designed for use with a diamond crystal (200-30000 cm-1) to measure the reflectance of solid and liquid samples. The spectra can be transformed by recording the absorbance as a function of wavelength. The concentrations can be calculated from absorbance measurements using a calibration curve. The interferogram is equipped with KBr beam splitter and DTGS/KBr detector. 12

13. Preparation of standard caffeine: Standard solutions of caffeine containing 0,2,4,6,8 and 10 mg in chloroform were pipette separately in 100 mg dry KBr. The standard samples were mixed, crushed and dried under N 2 gas to remove traces of chloroform. Preparation of coffee samples: Solid coffee samples finely powdered were used directly for analysis Conditions of measurements : Wave numbers 400-4000 cm -1 , Number of scans 32 , Resolution 4 cm -1 . Preparation of Samples 13

14. Determination of caffeine and active ingredient in different coffee samples by FTIR Overlain FTIR spectra FTIR/ATR spectra of solid samples of pure caffeine and coffee samples from top to bottom, Arabian coffee blend, roasted coffee blend, Instant coffee and caffeine standard respectively 14

15. Tga Analysis of caffeine samples Above demonstrates the thermograms of the samples with percentages loss of masses caused by heat flow through coffee samples. 15

16. Conclusion The aim of the comparative analysis has been focused on coffee samples containing spices to know their effect on caffeine. The results showed the absence of any chemical effect on the caffeine content of coffee samples as exhibited in the FTIR/ATR spectra. The purity of caffeine in Arabian coffee and roasted was maintained in the thermogram whereas the Instant coffee showed signs of impurity in the thermogram. Finally , the presence of spices in Arabian coffee and roasted home made coffee offers curative benefits and preventive medication as presented in the introduction which is good for the health of coffee consumers. 16

17. Reference International Journal of Engineering and Applied Sciences (IJEAS) ISSN: 2394-3661, Volume-2, Issue-12, December 2015 (by Mohammad A. Abdulla) Hindawi Publishing Corporation International Journal of Analytical Chemistry Volume 2015, Article ID 170239, 7 pages http://dx.doi.org/10.1155/2015/170239 (by Showkat Ahmad Bhawani, SimSiong Fong,and Mohammad Nassir Mohammad Ibrahim) https://www.hindawi.com/journals/ijac/2015/170239/ International Journal of Research In Science & Engineering e-ISSN: 2394-8299Volume:2 Issue:6 p-ISSN: 2394-8280 https://www.ncbi.nlm.nih.gov/m/pubmed/1658821/ https://en.m.wikipedia.org/wiki/Murexide_test https://www.mdpi.com/1420-3049/21/8/974/ html 17