4.1) Parameters and sampling frequency
|
|
- Camron Merritt
- 5 years ago
- Views:
Transcription
1 4.1) Parameters and sampling frequency At the height of two to three meters, fully expanded mature leaves were collected from each plant in the polythene bags and transported to the laboratory. The leaf samples were collected on seasonal basis and this frequency was strictly maintained throughout the year (November 2009 to October 2010). The following investigations were carried out in all the five plants Ficus religiosa, Ficus benghalensis, Ficus glomerata, Azadirachta indica and Polyalthia longifolia. 49 Dust fall on leaves was studied seasonally (winter, summer and rainy).for photosynthetic study the chlorophyll pigments (Chlorophyll a, chlorophyll b, and total chlorophyll) were studied and biochemical changes in leaves (Starch, phenols, and sugars- total, reducing and non-reducing) were studied on seasonal basis. Air pollution tolerance index and enzymatic activities i.e protease, catalase, peroxidase and invertase were studied in winter season. Experiment Physiological Metabolism Dustfall Chlorophyll APTI Biochemical Enzymatic
2 Biochemical Enzymatic Sugars Phenol Starch Protease Oxidase Invertase Total Reducing Non-reducing Catalase Peroxidase ) Method of measuring dust fall, chlorophyll pigments (total, a, and b) and air pollution tolerance index (APTI). A: Measurement of dust falls on the leaves From each plant, ten matured leaves were collected in the separate polythene bags during winter, summer and rainy from November 2009 to October Leaves were collected at the height of three to four meters from all the sites. For dust fall measurement, the method of Dry technique described by Das and Pattanayak (1997) was followed. In this technique first the intact leaf was weighted (in mg) then dust particulates from leaf surfaces were gently collected with the help of camel hair brushes and the weight of leaf was measured again. The amount of dust deposition in mg/cm 2 was calculaed as:- Weight of intact leaf- initial weight of leaf Dust content (mg/cm 2 ) = Total surface area of leaf (cm 2 )
3 B: Measurement of chlorophyll pigments The chlorophyll pigments in the leaves were estimated following the method of Arnon (1949). The fully expanded leaves from all the sites were collected in the poly-thene bags and transported to the laboratory. The leaves were washed out thoroughly with distilled water. Three replicates were used for each plant. 51 Weighted fresh leaf material was homogenized and extracted thrice in chilled 80% acetone (v/v). The volume of the acetone extract was made up to a known one and the optical density was read at 645nm and 663nm wavelengths on a spectrophotometer. The concentration of the chlorophyll pigments was calculated using the following formula and the results are expressed in mg/g fresh weight. Chlorophyll a = [(12.7 X OD at 663) (2.69 X OD at 645)] X dilution factor Chlorophyll b = [(22.9 X OD at 645) (4.68 X OD at 663)] X dilution factor Total chlorophyll = [(20.2 X OD at 645) (8.02 X OD at 663)] X dilution factor. C: Measurement of Air Pollution Tolerance Index Air pollution tolerance index (APTI) was determined by the method given by Singh and Rao, The samples were estimated for Leaf-extract ph (Singh and Rao, 1983), relative moisture content (Wealtherly, 1965), total chlorophyll (Arnon, 1949) and ascorbic acid (Abida begum et al., 2010). The fully expanded leaves from all the sites were collected in the poly-thene bags and transported to the laboratory. The leaves were washed out thoroughly with distilled water. Three replicates were used for each plant.
4 Estimation of Leaf-extract ph (Singh and Rao, 1983): 0.5 g of leaf material was ground to paste and dissolved in 50 ml of distilled water and Leaf-extract ph was measured by using calibrated digital ph meter. Relative moisture content (Wealtherly, 1965): Estimation of relative moisture content: Fresh leaf samples collected from the study area and were brought immediately to the laboratory and washed thoroughly. The excess water was removed with the help of filter paper. The initial weight of samples were taken (W1 g) and kept in oven at 600 o C until constant weight was obtained and the final weight was taken (W2 g). 52 Total Chlorophyll content was measured by the method of Arnon (1949) as mentioned above. Ascorbic acid content (AA) (mg/g) was measured using spectrophotemetric method. 1 g of the fresh foliage was put in a test-tube, 4 ml oxalic acid - EDTA extracting solution was added, then 1 ml of orthophosphoric acid and then 1 ml 5% tetraoxosulphate(vi) acid added to this mixture, 2 ml of ammonium molybdate was added and then 3 ml of water. The solution was then allowed to stand for 15 minutes. After which the absorbance at 760 nm was measured with a spectrophotometer (Abida Begum and Krishna, 2010). APTI given as: APTI = [AA (T + P) + R] 10 (Where AA is the ascorbic acid in mg/g, T is the total chlorophyll in mg/g, P is ph of leaf sample and R is relative water content in mg/g).
5 4.3) Method of measuring sugar (total, reducing and nonreducing), phenol and starch. A: Measurement of Sugars (Nelson, 1944) Principle: Monosaccharide readily reduces oxidizing agents such as Ferric cyanide, hydrogen peroxide or cupric ions (Cu ++ ). In such reactions the sugar is oxidized at carbonyl group and the oxidizing agent becomes reduced glucose or other sugars capable of reducing oxidizing agents are called reducing sugars. Thus by measuring the amount of oxidizing agent that is reduced by a sugar solution, it is possible to estimate the sugar concentration. The method involves the reduction of cupric ions (Cu ++ ) to cuprous ions (Cu + ) which in alkaline solution and forms yellow cuprous hydroxide, which in turn is converted by heat of the reaction to insoluble red cuprous oxide (Cu 2 O). 53 The amount of Cu 2 O formed can be increased by adding arsenomolybdic acid which in turn is reduced to lower oxides of molybdenum by Cu 2 O. The coloured complex produced is known as molybdenum blue. The intensity of the colour is related to the concentration of the reducing sugars in the sample. Procedure: 100mg plant material was weighed and homogenate with 10ml 80% ethanol. It was centrifuged for 10rpm for 10minutes. Supernatant 1 was collected; while 10 ml 80% ethanol was added again to the residue, centrifuge it and supernatant 2 was mixed with supernatant 1. Residue was discarded. Total sugars: To 1ml alcoholic aliquot, 1ml 1N H 2 SO 4 was added and heated at 49 0 C in water bath for 30 minutes for hydrolysis of the mixture. 1-2 drop of methyl red indicator was added. 1N NaOH was added drop wise for the neutralization (colour was to yellow from pink). 1ml Nelson Somogyi s reagent was added to it and the tube was kept in boiling water bath for 20 minutes. After cooling of the test tube, 1ml arsenomolybdate was added and final
6 volume was made up to 20ml with DW. O.D. was noted at 540nm. Blank was prepared in the same manner. Reducing sugars: To 1ml alcoholic aliquot, Nelson Somogyi s reagent was added and kept in boiling waterbath for 20min. After cooling of the test tube, 1ml arsenomolybdate was added and final volume was made upto 20ml with DW. O.D. was noted at 540nm. Blank was prepared in the same manner. Non-reducing sugar= Total sugar Reducing sugar. The result was expressed as mg/gm plant material. 54 Preparation of reagents: 1.80%Ethanol: 80ml Ethanol was diluted up to 100ml DW. 2.1N Sulphuric acid (H 2 SO 4 ): 2.77ml conc. H 2 SO 4 (95-98%) was diluted up to 100 ml with DW N Sodium hydroxide (NaOH): 4gm NaOH was dissolved up to 100ml with DW. 4. Methyl red indicator (1N): 0.1gm Methyl red powder was dissolved in 5ml 0.02M NaOH and final volume was made up to 250ml with DW. 5. Nelson Somogyi s reagent: Nelson A: 12.5gm Na 2 CO 3, 12.5gm Na-K-tartrate, 10gm NaHCO 3, 100gm Na 2 SO 4 were dissolved one by one and final volume made up to 50ml with DW. Nelson B: 15gm CuSO 4.7H 2 O dissolved up to 100ml with DW. Nelson Somogyi s reagent: 50ml Nelson A and 1ml Nelson B were mixed. 6. Arsenomolybdate reagent: 25gm Ammonium molybdate was dissolved in 450ml DW, 21ml conc. H 2 SO 4 was added to it. 3gm Sodium arsenate was dissolved in 25ml DW and both solutions were mixed. It was incubated at 35 0 C for overnight before use of it.
7 CHART- 4.3(a): FLOW CHART FOR TOTAL SUGARS AND REDUCING SUGARS 100 mg plant material was weighed Homogenized with 10 ml 80% ethanol Centrifuged at 5,000-10,000rpm for 10 minutes 55 Supernatant I Residue +10ml 80% ethanol Centrifuged at g for 10 minutes Supernatant II Residue Supernatant I+II Total sugar Reducing sugar 1 ml aliquot + 1ml aliquot + 1 ml 1 N H 2 SO 4 1ml Nelson Somogyi s reagent Incubated in water bath at 49 0 C for 30 min. Add 1-2 drop of methyl red indicator Add 1N NaOH drop wise for neutralization (Colour change Pink to Yellow) Incubated in boiling water bath for 20min Add 1ml Arsenomolybdate Final vol. made upto 20ml with D.W OD was noted at 540 nm 1ml Nelson Somogyi s reagent Incubate for 20mins in boiling water bath Add 1ml Arsenomolybdate Final volume was made up to 20ml with D.W. O.D. at 540nm.
8 The readings were compared with a standard which was prepared by using different concentration of glucose. The results were expressed as mg/g plant material. B: Measurement of Total phenols (Bray et al., 1954) Principle: Estimation of phenols using Folin- Ciocalteu s reagent is based on the reaction between phenols and an oxidizing agent phosphomolybdate which results in the formation of a blue complex (Bray et al., 1954). 56 Procedure: 100mg plant material was weighed and homogenate with 10ml 80% ethanol. It was centrifuged at ,000rpm for 10minutes. Supernatant 1 was collected; while 10 ml 80% ethanol was added again to the residue, centrifuge it and supernatant 2 was mixed with supernatant 1 and used for estimation. Residue was discarded.1ml alcoholic aliquot was mixed with 1ml 20% Na 2 CO 3 and 0.5ml Folin-Ciocalteau s reagent. It was boiled for 10minutes at C in water bath. Final volume was made up to 20ml with DW and O.D. was noted at 660nm. PPT were filtered or centrifuged before reading. Blank was prepared in the same manner. The result was expressed as mg/gm plant material. Preparation of reagents: 1. 80%Ethanol: 80ml Ethanol was diluted up to 100ml DW % Sodium carbonate (Na 2 CO 3 ):20gm Na 2 CO 3 was dissolved into 100ml DW. 2. Folin-ciocalteau s reagent (1N): Commercially available reagent (2N) was diluted with an equal volume of DW.
9 CHART4.3 (b): FLOW CHART FOR TOTAL PHENOLS 100mg plant material crushed in 10ml 80% ethanol Centrifuge at ,000 g for 10mins Supernatant 1 residue + 10ml 80% ethanol 57 Centrifuge Supernatant 2 + Supernatant 1 residue discarded 1ml aliquot + 1ml 20% Na 2 CO ml Folin-Ciocalteau s reagent Boil in water bath for 10mins Final volume made up to 20ml with DW Ppt. filtered O.D. at 660nm The readings were compared with a standard which was prepared by using tannic acid. The results were expressed as mg/g plant material.
10 C: Measurement of Starch (Chinoy, 1939) Principle: The plant material is treated with aqueous sodium hydroxide in the cold to dissolve the starch. This dissolved starch reacts with I 2 KI and gives a coloured product and the starch content is determined by calculated with the standard curve. Procedure: 100mg plant material was weighed and homogenate with 10ml 80% ethanol. It was centrifuged for 10minutes. Supernatant 1 was collected; while 10 ml 80% ethanol was added again to the residue centrifuge it and supernatant 2 was mixed with supernatant 1 and removed. Residue was used for starch estimation. The residue was dissolved in 20ml 0.7% KOH and boiled for gelatinization for 40 minutes. It was centrifuged after cooling and 1ml aliquot (Supernatant), 0.5ml 20% acetic acid; 1ml citrate buffer (0.05M, ph 5.0) and 1ml I 2 KI were added and incubated at room temperature for 10minutes. O.D. was taken at 600nm. Blank was prepared in the same manner. The result was expressed as mg/gm plant material. 58 Preparation of reagents: 1.80%Ethanol: 80ml Ethanol was diluted up to 100ml DW % Potassium hydroxide (KOH): 700 mg KOH was dissolved into 100ml DW %acetic acid: 20ml glacial acetic acid was diluted up to 100ml with DW. 4. I 2 KI Solution: 200mg iodine crystal and 2gm KI were dissolved up to 100ml with DW. 5. Citrate buffer: (0.05M, ph 5.0) Citrate X: 0.1M Citric acid (2.19gm Citric acid was dissolved into 100ml DW.) Citrate Y: 0.1M Sodium Citrate (2.94gm Sodium Citrate was dissolved into 100ml DW.) Citrate buffer: 20.5ml Citrate X and 29.5ml Citrate Y were dissolved up to 100ml with DW.
11 CHART4.3(c): FLOW CHART FOR STARCH 100mg plant material crushed in 10ml 80% ethanol Centrifuge at ,000 g for 10mins Supernatant 1 residue + 10ml 80% ethanol 59 Centrifuge at 5,000-10,000g for 10min Supernatant 2 + Supernatant 1 residue Discarded Dissolved in 20ml 0.7% KOH and boiled for gelatinization for 40mins. Cooled and centrifuged 1ml aliquot 0.5ml 20% acetic acid + 1ml citrate buffer + 1ml I 2 KI Incubate at room temperature for 10mins O.D. at 600nm The readings were compared with a standard which was prepared by using starch. The results were expressed as mg/g plant material.
12 4.4) Method of measuring enzyme activity (Protease, Catalase, Peroxide, Invertase). Method for enzyme extraction: Grind 1gm plant material in 10ml phosphate buffer. Centrifuge the extract at 10,000 rpm for 15minutes at 4 0 C refrigerated centrifuge. Preparation of reagents: 1. Phosphate buffer (0.1M, ph=7): Phosphate A: 0.2 M dibasic sodium phosphate (35.61g Na 2 HPO 4.7H 2 O was dissolved up to 1,000ml with DW. Phosphate B: 0.2 M monobasic sodium phosphate (31.21g NaH 2 PO 4.2H 2 O was dissolved up to 1,000ml with DW.) Phosphate buffer: 61ml Phosphate A and 39ml Phosphate B were diluted up to 100ml DW. 60 CHART: FLOW CHART FOR ENZYME EXTRACTION Take 1gm plant material Grind it in 10ml phosphate buffer Centrifuge at 10,000 rpm for 15mins at 4 0 C Use supernatant for enzyme activity. A: Measurement of Protease activity (Cruz et al., 1970) Principle: Protease hydrolyses protein into its constituent amino-acids. By estimating the amount of protein hydrolyzed in a solution, the activity of the protease enzyme can be known.
13 The Folin- Lowry assay used for protein estimation was used here for determining the activity of the protease enzyme. Protein reacts with Folin reagent to give a coloured complex. The intensity of the colour depends upon the amount and type of aromatic amino-acid produced due to hydrolyses by the enzyme. The type of amino-acid varies for different proteins. Procedure: 3 test tube, 1ml enzyme aliquot, 1ml phosphate buffer (0.2 M, ph=7.0 and 1ml 1% casein solution were mixed. It was incubated at room temperature for 60 min. 1ml 20% Trichloro acetic acid (TCA) was added to it. Standard tube had 1ml DW, 1ml Phosphate buffer, 1ml Trichloro acetic acid (TCA) and 1ml casein. Blank has 2ml DW, 1ml phosphate buffer and 1ml Trichloro acetic acid (TCA). All test tubes were incubated at room temperature for 60 minutes. All the 3 test tubes were centrifuged and residue was discarded. 1ml aliquot (supernatant) was mixed with 5ml Lowry C and incubated at room temperature for 10 min. 0.5 ml Folin - Ciocalteau s reagent was added in each and again incubated for 10 minutes at room temperature. O.D of blank was related with zero setting, standard gave the reading of casein, while test gave reading of reduced amount of protein at 600nm. Subtraction of control and test gave protease activity. Result was expressed as mg/protein reduced/gm plant material. Preparation of reagent: 1. Phosphate buffer (0.2M, ph=7): 61 Phosphate A: 0.2M Monobasic sodium phosphate. Phosphate B: 0.2M Dibasic sodium phosphate. Phosphate buffer: 39ml phosphate A was mixed with 61ml phosphate B. 2. 1% casein solution: 1gm casein was dissolved in 5ml 1N NaOH and final volume was made up to 100ml with DW % Trichloro acetic acid (TCA): 20gm TCA was dissolved in 100ml DW. 4. Lowry s reagent:
14 Lowry A: 2% Na 2 CO 3 in 0.1N NaOH. Lowry B: 0.5% CuSO 4 in 1% of Na-K-tartrate. Lowry C: 50ml Lowry A was mixed with 1ml Lowry B. 5. Folin-Ciocalteaus reagent (1N): Commercially available reagent (2N) was diluted with an equal volume of DW. CHART4.4 (a): FLOW CHART FOR PROTEASE Test Standard Blank 62 1ml enzyme extract 1ml DW 2ml DW 1ml Phosphate Buffer 1ml 1% Casein Incubated for 1hr 1ml 20% TCA (Trichloro Acetic Acid) Incubate for 1hr. Centrifuge Supernatant 1ml Supernatant Residue (Discarded) 5ml Lowry Reagent
15 Incubated for 10minutes Add 0.5ml Folin Reagent Incubated for 10min O.D. at 600 The readings were compared with a standard which was prepared by using different concentration of tyrosine. The results were expressed in mg tyrosine liberated/hr/mg protein. 63 B: Measurement of Catalase activity (Chance and Maehly, 1955) Principle: The enzyme catalase is an endogenous antioxidant present in all aerobic cells helping to facilitate the removal of hydrogen peroxide. The enzyme consists of 4 subunits of the same size, each of which contains a heme active site to accelerate the decomposition of H 2 O 2 to water and oxygen. Procedure: This activity was done by titration method. Reaction, mixture was prepared by mixing 3ml of phosphate buffer (0.1M, ph= 6.8), 1ml 0.1M H 2 O 2 and 1ml enzyme aliquot. It was incubated at room temperature for 1min. Futher reaction was stopped by addition of 10ml 20% H 2 SO 4. This mixture was titrated against 0.01N KMnO 4 to estimate the residual H 2 O 2 until a faint pink colour persisted for at least 15secs. This enzyme activity was expressed as amount of enzyme break down by H 2 O 2 /min/gm plant material. Preparation of reagent:- 1) Phosphate buffer (0.1M, ph=6.8): Phosphate A: 0.2M Monobasic sodium phosphate.
16 Phosphate B: 0.2M Dibasic sodium phosphate. Phosphate buffer: 51ml phosphate A and 49ml phosphate B were diluted upto 200ml DW. 2) 2% Sulphuric acid (H 2 SO 4 ): 2ml conc. H 2 SO 4 was diluted upto 100ml DW. 3) 0.01N Potassium permanganate (KMnO 4 ): mg KMnO 4 was dissolved upto 100ml DW. 64 4) 0.1M Hydrogen peroxide (H 2 O 2 ): 3.041ml H 2 O 2 (100v/v, 30%) was diluted upto 1000ml DW. CHART4.4(b): FLOW CHART FOR CATALASE 1ml Enzyme aliquot + 3ml phosphate buffer (0.1M, ph=6.8) + 1ml 0.1MH 2 O 2 Incubated at room temperature for 1min Added 10ml 2% H 2 SO 4 Titrated against 0.01N KMnO 4 to estimate the residual H 2 O 2 until a faint pink colour persisted for at least 15sec Expressed enzyme activity as amount of ml enzyme broke Down by H 2 O 2 /minute/gm plant material.
17 C: Measurement of Peroxidase activity (George, 1955) Procedure: 1ml enzyme aliquot was mixed with 1ml phosphate buffer (0.1M, ph= 6.4) and 1ml 20mM guaiacol. Optical density was noted at 420nm. 0.5ml H 2 O 2 was added and reading was noted after 2 mins. Calculation was done expressed as O.D difference/minute/gm plant material. Blank was prepared in the same manner. Preparation of the reagent: 1) Phosphate buffer (0.1 M, 6.4= ph): Phosphate A: 0.2 M monobasic sodium phosphate (27.8g NaH 2 PO 4.2H 2 O was dissolved upto 1,000ml with DW. Phosphate B: 0.2 M dibasic sodium phosphate (53.65g Na 2 HPO 4. 7H 2 O was dissolved upto 1,000 with DW. Phosphate buffer: 73.5ml Phosphate A and 23.5ml Phosphate B were diluted upto 200ml DW. 2) 20mM Guaiacol: 0.22ml Guaiacol was upto 100ml with DW. 65 CHART4.4(c): FLOW CHART FOR PEROXIDASE Test Blank 1ml Enzyme + 1ml DW + 1ml Phosphate buffer + 1ml Phosphate buffer + 1ml Guaiacol (20mM) 1ml Guaiacol (20 mm) O.D. at 420 nm 0.5ml H 2 O 2 Incubate for 2min O.D. read at 420nm
18 The results of enzyme activity were expressed as O.D. difference/min/mg protein. D: Measurement of Invertase (Hatch and Glasziou, 1963) Principle:Invertase is an enzyme which hydrolyses disaccharides into monosaccharide. Sucrose is the substance that gets reduced to monosaccharide that is reducing sugar which can be produces by invertase and can also be measured. Monosaccharide acts as an oxidizing agent and oxidizes Cu ++. The sugar that reduces the oxidizing agent is known as reducing sugar. The method involves reduction of cupric ions (Cu ++ ) into cuprous ion (Cu + ) which is alkaline in nature and forms yellow cuprous hydroxide, which in turn is converted by heat of the reaction to insoluble red cuprous oxide (Cu 2 O). The amount of Cu 2 O is dissolved in arsenomolybdate and forms a coloured complex. The developed colour is related to the concentration of the reducing sugars and is measured using the spectrometer. 66 Procedure: 3 test tubes were taken for this activity (1) test (2) control (3) blank.1ml enzyme aliquot, 1ml 0.1M sucrose solution in citrate buffer (0.1M, ph=5.4) and 1ml citrate buffer were added to the test tube.1ml DW, 1ml sucrose solution and 1ml citrate buffer were added to the control tube.2ml DW and 1ml citrate buffer were added to the blank tube. All test tubes were incubated at room temperature for 60minutes. After the incubation, 2ml absolute alcohol and 2ml 5% Sodium sulphate (Na 2 SO 4 ) were added in all 3t.t. All test tubes were again incubated in boiling water bath for removal or evaporation of alcohol (10-20 minutes). Impurities were filtered or centrifuged and each test tube had left 5ml solution. 1ml solution aliquot and 1ml Nelson somogyi s reagent were mixed and kept in boiling water bath for 20 minutes. 1ml arsenomolybdate was added to each after cooling. Final volume was made
19 up to 20ml with DW. O.D. was noted at 540nm.Blank was related with zero setting, control gave the reading of sucrose, while test gave reading of reduced sucrose. Subtraction of control and test gave Invertase activity. Result was expressed as glucose reduced/gm plant material. Preparation of the reagent: 1. Citrate buffer (0.1 M, ph=5.4): Citrate A: 0.1M citric acid (21.01gm citric acid was dissolved up to 1000ml with DW.) Citrate B: 0.1M Sodium citrate (29.41gm Sodium citrate was dissolved up to 1000ml with DW.) Citrate buffer: 16ml citrate A and 34ml citrate B were mixed M sucrose in citrate buffer: 3.42gm of sucrose was dissolved up to 1000ml with citrate buffer (0.1 M, PH-5.4) 3. Sodium sulphate (Na 2 SO 4 ) (5%): 5g Na 2 SO 4 was dissolved in 100ml D.W. 4. Nelson somogyi s reagent: Nelson A : 12.5 gm Na 2 CO 3, 12.5 gm Na-k tartrate, 10g NaHCO 3, 100g Na 2 SO 4 were dissolved one by one and final volume was made up to 500 ml DW. Nelson B :15g CuSO 4.7H 2 O was dissolved up to 100ml with DW. Nelson Somogyi s reagent: 50 ml Nelson A and 2ml Nelson B was mixed. 5. Arsenomolybdate reagent:- 25gm ammonium molybdate was dissolved in 450ml DW, 21 ml concentrated H 2 SO 4 was added to it. 3g sodium arsenate was dissolved in 25 ml DW and both solutions were mixed. It was incubated at 37 0 c over night before use of it. 67
20 CHART4.4 (d): FLOW CHART FOR INVERTASE Test Standard Blank 1ml enzyme extract 1ml DW 2ml DW 1ml Sucrose 68 1ml citrate Buffer (0.5 M, ph 5.4) Incubate for 1 hr 2ml alcohol + 2ml 5% Sodium Sulphate (Na 2 SO 4 ) Incubate in boiling water bath for evaporation of alcohol (10-20 minutes) Impurities were filtered or centrifuged 1ml solution aliquot + 1ml Nelson Somogyi s reagent Incubation in boiling water bath for 20 minutes Add 1ml AMR (Arsenomolybdate reagent) Prepare final volume 20ml with D.W. O.D. at 540nm
21 The O.D was compared with a standard which was prepared using different concentration of glucose. The results of enzyme activity were expressed as mg glucose released/hr/mg protein. 4.5 STATISTICAL RESULT It is essential to understand the relationship between different parameter when the study is completed Relationship between the parameters (Correlation) Any relationship between the two variable is known as correlation. If one variable increases or decreases with a corresponding increase or decrease of the other variable, a direct positive correlation exists between the two variables. If one variable decrease with an increase in the other variable, then there is a negative or inverse correlation. There are two different methods to study correlation Graphic method It is the simplest method of showing the relationship between two variable. In this one variable is represented on X-axis and other variable on Y- axis on graph paper. Data corresponding to X and Y axis were plotted in form of dots. And then estimated lines joining first and last points was drawn on the graph paper to find out correlation. Correlation coefficient The graphic method indicates the existence of a correlation. But it is not possible to calculate the extent or degree of relationship using these graph. So,this was calculated by using following formula
22 r = (dx. dy) (dx) 2. (dy) 2 Where, r is the correlation coefficient, x and y are the two variable dx is the deviation from the x-mean of the x variable, 70 dy is the deviation from the y mean of the y variable, (dx. dy) is the sum of the products of the deviations, (dx) 2 is the sum of the squares of the deviations of the x variable, (dy) 2 is the sum of the squares of the deviations of the y variable,
Aim: To study the effect of ph on the action of salivary amylase. NCERT
Exercise 28 Aim: To study the effect of ph on the action of salivary amylase. Principle: Optimal activity for most of the enzymes is generally observed between ph 5.0 and 9.0. However, a few enzymes, e.g.,
More informationEXPERIMENT 4 DETERMINATION OF REDUCING SUGARS, TOTAL REDUCING SUGARS, SUCROSE AND STARCH
Practical Manual Food Chemistry and Physiology EXPERIMENT 4 DETERMINATION OF REDUCING SUGARS, TOTAL REDUCING SUGARS, SUCROSE AND STARCH Structure 4.1 Introduction Objectives 4.2 Experiment 4a: Reducing
More informationMANUAL OF RESEARCH MiiTHODS FOR CRUSTACEAN BIOCHEMISTRY AND PHYSIOLOGY
f M ' CMFRI SPECIAL PUBLICATION Number 7 MANUAL OF RESEARCH MiiTHODS FOR CRUSTACEAN BIOCHEMISTRY AND PHYSIOLOGY ISSIH;(! on Hie occasion of the Wotkshop ott CRUSTACEAN BIOOHfcMISTHY AND PHYSIOLOGY jointly
More informationCHAPTER 3: MATERIALS AND METHODS
CHAPTER 3: MATERIALS AND METHODS Materials and Methods. The from different husks, fruits, and vegetables peels were estimated quantitatively by the following volumetric procedure s such as Bertrand s,
More informationPurity Tests for Modified Starches
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Purity Tests for Modified Starches This monograph was also published in: Compendium
More informationFor example, monosaccharides such as glucose are polar and soluble in water, whereas lipids are nonpolar and insoluble in water.
Biology 4A Laboratory Biologically Important Molecules Objectives Perform tests to detect the presence of carbohydrates, lipids, proteins, and nucleic acids Recognize the importance of a control in a biochemical
More informationQUANTITATIVE TEST (CHEMICAL) FOR SUGARS IN SUGARCANE. Talha Saeed. Faisal Iftikhar. Mam AMMARA AINEE
Assignment title QUANTITATIVE TEST (CHEMICAL) FOR SUGARS IN SUGARCANE Submitted by Subject Talha Saeed Roll # 37 Faisal Iftikhar Roll # 18 B.Sc. (Hons) Food Science and Technology 6 th Semester (Regular)
More informationHiPer Carbohydrates Estimation Teaching Kit (Quantitative)
HiPer Carbohydrates Estimation Teaching Kit (Quantitative) Product Code: HTBC003 Number of experiments that can be performed: 10 Duration of Experiment Protocol DNSA Method :1 hour Phenol Sulphuric Acid
More informationExperiment 1. Isolation of Glycogen from rat Liver
Experiment 1 Isolation of Glycogen from rat Liver Figure 35: FIG-2, Liver, PAS, 100x. Note the presence of a few scattered glycogen granules (GG). Objective To illustrate the method for isolating glycogen.
More informationMIXED XYLANASE, β-glucanase ENZYME PREPARATION, produced by a strain of HUMICOLA INSOLENS
MIXED XYLANASE, β-glucanase ENZYME PREPARATION, produced by a strain of HUMICOLA INSOLENS New specifications prepared at the 61st JECFA (2003) and published in FNP 52 Add 11 (2003). An ADI not specified
More information4. Determination of fat content (AOAC, 2000) Reagents
94 ANALYTICAL METHODS 1. Determination of moisture content (AOAC, 2000) 1. Dry the empty dish and lid in the oven at 105 C for 3 h and transfer to desiccator to cool. Weigh the empty dish and lid. 2. Weigh
More informationG/LITRE 5.0 g KOH g 0.5 g 0.05 g 0.01 g MgS047H20 NaCl CaCl2
A P P E N D IX -V III COMPOSITION OF USED MEDIA AND CHEMICAL REAGENTS 1. NITROGEN FREE BROMOTHYMOL BLUE (NFB) MEDIUM Dobereiner et al (1976) Same media was also used to check the effect of temperature
More informationMATERIALS AND METHOD
Chapter - 3 Histomorphology, Ecology and Biochemistry of leaf galls of Ficus glomerata Roxb. induced by Pauropsylla depressa Crawford. MATERIALS AND METHOD Field observations were confined in Saharanpur
More informationAZO-XYLAN (BIRCHWOOD)
ASSAY OF endo-1,4-ß-xylanase using AZO-XYLAN (BIRCHWOOD) S-AXBP S-AXBL 10/07 Megazyme International Ireland 2007 PRINCIPLE: This assay procedure is specific for endo-1,4-ß-d-xylanase activity. On incubation
More informationName: Period: Date: Testing for Biological Macromolecules Lab
Testing for Biological Macromolecules Lab Introduction: All living organisms are composed of various types of organic molecules, such as carbohydrates, starches, proteins, lipids and nucleic acids. These
More informationWork-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples:
Dr. Sanjeeva Srivastava IIT Bombay Work-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples: Sample preparation for serum proteome analysis Sample
More informationTECHNICAL BULLETIN METHOD 1: DETERMINATION OF TOTAL DIETARY FIBRE
TOTAL DIETARY FIBER KIT Cat N 32 v.3210051 TECHNICAL BULLETIN METHOD 1: DETERMINATION OF TOTAL DIETARY FIBRE Introduction This procedure for the determination of total dietary fiber is based on the method
More informationΒ-FRUCTOFURANOSIDASE ENZYME
KINETICS ANALYSIS OF Β-FRUCTOFURANOSIDASE ENZYME 2-The effects of enzyme concentration on the rate of an enzyme catalyzed reaction. Systematic names and numbers β-fructofuranosidase (EC 3.2.1.26) Reactions
More informationE55A GELATIN, GELLING GRADE Gelatina
00-0PDG.pdf 0 0 0 0 EA GELATIN, GELLING GRADE Gelatina DEFINITION Purified protein obtained from collagen of animals (including fish and poultry) by partial alkaline and/or acid hydrolysis, by enzymatic
More information5. BIOCHEMICAL COMPOSITION AND FOOD VALUE OF RIBBON FISH L. SAVALA
5. BIOCHEMICAL COMPOSITION AND FOOD VALUE OF RIBBON FISH L. SAVALA During present study, sixty specimens of fresh L. savala ranging from 200 to 600 mm of total length were collected from Baithkol, Majali
More informationMATERIAL AND METHODS
MATERIAL AND METHODS Material and Methods Glucose induced cataract was chosen as a model for the present study. A total of 210 fresh goat lenses were analyzed. Sample Collection: Goat eyeballs were obtained
More informationInt. J. Pharm. Sci. Rev. Res., 14(2), 2012; nᵒ 22, COMPARATIVE CARBOHYDRATES STATUS IN LEAF DEVELOPMENTAL STAGES OF CLEOME SPECIES
Research Article COMPARATIVE CARBOHYDRATES STATUS IN LEAF DEVELOPMENTAL STAGES OF CLEOME SPECIES Vishal T. Aparadh*, B. A. Karadge. Department of Botany, Shivaji University, Kolhapur. (M.S.) India. 416
More informationEnzymatic Assay of POLYGALACTURONASE (EC )
PRINCIPLE: Polygalacturonic Acid + H 2 O PG > Reducing Sugars Abbreviations: PG = Polygalacturonase CONDITIONS: T = 30 C, ph 5.0, A 540nm, Light path = 1 cm METHOD: Colorimetric REAGENTS: A. 50 mm Sodium
More informationAPPENDIX Heparin 2 mg heparin was dissolved in 0.9 % NaCl (10 ml). 200 µl of heparin was added to each 1 ml of blood to prevent coagulation.
APPENDIX 1 Preparation of reagents 1.1. Preparation of dosing solution Nonylphenol 15 mg of Nonylphenol was dissolved in olive oil (10 ml) and used as stock solution. The stock solution was serially diluted
More informationINTERNATIONAL ŒNOLOGICAL CODEX. DETERMINATION OF BETA-GLUCANASE (ß 1-3, ß 1-6) ACTIVITY IN ENZYME PREPARATIONS (Oeno 340/2010, Oeno )
DETERMINATION OF BETA-GLUCANASE (ß 1-3, ß 1-6) ACTIVITY IN ENZYME PREPARATIONS (Oeno 340/2010, Oeno 488-2013) General specifications These enzymatic activities are usually present within a complex enzymatic
More informationIODOMETRIC TITRATION
IODOMETRIC TITRATION Oxidizing agents In most iodometric titrations, when an excess of iodide ion is present, the tri-iodide ion is formed: I + I - I 3 - Since iodine is readily soluble in a solution of
More informationEnzymatic Assay of ß-GLUCOSIDASE (EC )
PRINCIPLE: ß-D-Glucoside + H 2 O ß-Glucosidase > D-Glucose + an Alcohol CONDITIONS: T = 37 C, ph = 5.0, A 540nm, Light path = 1 cm METHOD: Colorimetric 1 REAGENTS: A. 100 mm Sodium Acetate Buffer, ph 5.0
More informationResidue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016.
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Aspartame This monograph was also published in: Compendium of Food Additive
More information-Glucan (mixed linkage), colorimetric method
-Glucan (mixed linkage), colorimetric method Catalogue number: AK0027, 00 tests Introduction -Glucans are common components in cereals, bacteria, yeasts and mushrooms. Mixed linkage -glucans are naturally
More informationLACTOSE/ SUCROSE/D-GLUCOSE
www.megazyme.com LACTOSE/ SUCROSE/D-GLUCOSE ASSAY PROCEDURE FOR THE MEASUREMENT OF LACTOSE, SUCROSE AND D-GLUCOSE IN FLOURS K-LACSU 06/15 (100 Assays of each per Kit) Megazyme International Ireland 2015
More informationResearch Article GALLIC ACID AND FLAVONOID ACTIVITIES OF AMARANTHUS GANGETICUS
ISSN 2395-3411 Available online at www.ijpacr.com 238 Research Article GALLIC ACID AND FLAVONOID ACTIVITIES OF AMARANTHUS GANGETICUS G. Jyoti Jain 1* and S. Ramachandra Setty 2 1 Department of Pharmacology,
More information(Writing model for laboratory note book)
Paper: Lab 50 Syllabus *************************************************************************** Experiment: Organic Qualitative analysis 1) Detection of elements (Nitrogen, Sulphur and halogens). 2)
More informationScreening of bacteria producing amylase and its immobilization: a selective approach By Debasish Mondal
Screening of bacteria producing amylase and its immobilization: a selective approach By Debasish Mondal Article Summary (In short - What is your article about Just 2 or 3 lines) Category: Bacillus sp produce
More information6 The chemistry of living organisms
Living organisms are composed of about 22 different chemical elements. These are combined to form a great variety of compounds. Six major elements make up almost 99% of the mass of the human body, as shown
More informationB. 1% (w/v) Salicin Substrate Solution (Salicin) (Prepare 50 ml in Reagent A using Salicin, Sigma Prod. No. S-0625.)
SIGMA QUALITY CONTROL TEST PROCEDURE (Q]\PDWLFÃ$VVD\ÃRIÃ */8&26,'$6( PRINCIPLE: 'Glucoside + H 2 O Glucosidase > D-Glucose + an Alcohol CONDITIONS: T = 37 C, ph = 5.0, A 540nm, Light path = 1 cm METHOD:
More informationHEMICELLULASE from ASPERGILLUS NIGER, var.
HEMICELLULASE from ASPERGILLUS NIGER, var. Prepared at the 55th JECFA (2000) and published in FNP 52 Add 8 (2000), superseding tentative specifications prepared at the 31st JECFA (1987) and published in
More information2. 2,4 Dinitro phenyl hydrazine (DNPH): I mm in 1N HCl. 5. Working standard: 1 in 20 dilution of the stock standard.
-1 Estimation of Alanine Transaminase (ALT) (Mohun and Cook, 1957) Reagents I. Buffered substrate: [100 mm phosphate buffer, 200mM DL-alanine; 2 mm 2-oxo glutarate.}- Dissolved 1.5 g di potassium hydrogen
More informationBIOL 347L Laboratory Three
Introduction BIOL 347L Laboratory Three Osmosis in potato and carrot samples Osmosis is the movement of water molecules through a selectively permeable membrane into a region of higher solute concentration,
More informationINTERNATIONAL ŒNOLOGICAL CODEX
DETERMINATION OF POLYGALACTURONASE ACTIVITY IN ENZYMATIC PREPARATIONS endo- and exo-polygalacturonase activities (PG) (EC. 3.2.1.15 CAS N 9032-75-1) (Oeno 10/2008; Oeno 364-2012) General specifications
More informationEnzymatic Assay of PROTEASE (EC )
Enzymatic Assay of PROTEASE PRINCIPLE: Hemoglobin + H 2 O Protease > Amino Acids CONDITIONS: T = 37 C, ph = 2.8, A 660nm, Light path = 1 cm METHOD: Colorimetric REAGENTS: A. 50 mm Potassium Phthalate Buffer,
More informationTHERMALLY OXIDIZED SOYA BEAN OIL interacted with MONO- and DIGLYCERIDES of FATTY ACIDS
THERMALLY OXIDIZED SOYA BEAN OIL interacted with MONO- and DIGLYCERIDES of FATTY ACIDS Prepared at the 39th JECFA (1992), published in FNP 52 Add 1 (1992). Metals and arsenic specifications revised at
More informationEnzyme Action: Testing Catalase Activity
Enzyme Action: Testing Catalase Activity LabQuest 6A Many organisms can decompose hydrogen peroxide (H 2 O 2 ) enzymatically. Enzymes are globular proteins, responsible for most of the chemical activities
More informationQualitative test of protein-lab2
1- Qualitative chemical reactions of amino acid protein functional groups: Certain functional groups in proteins can react to produce characteristically colored products. The color intensity of the product
More informationSPECIFICATION CONTINUED Glucose has two isomers, α-glucose and β-glucose, with structures:
alevelbiology.co.uk SPECIFICATION Monosaccharides are the monomers from which larger carbohydrates are made. Glucose, galactose and fructose are common monosaccharides. A condensation reaction between
More informationProcine sphingomyelin ELISA Kit
Procine sphingomyelin ELISA Kit For the quantitative in vitro determination of Procine sphingomyelin concentrations in serum - plasma - celiac fluid - tissue homogenate - body fluid FOR LABORATORY RESEARCH
More informationCELLULASE from PENICILLIUM FUNICULOSUM
CELLULASE from PENICILLIUM FUNICULOSUM Prepared at the 55th JECFA (2000) and published in FNP 52 Add 8 (2000), superseding tentative specifications prepared at the 31st JECFA (1987) and published in FNP
More informationOrganic Molecule Composition of Milk: Lab Investigation
Name: Organic Molecule Composition of Milk: Lab Investigation Introduction & Background Milk & milk products have been a major food source from earliest recorded history. Milk is a natural, nutritionally
More informationTHERMALLY OXIDIZED SOYA BEAN OIL
THERMALLY OXIDIZED SOYA BEAN OIL Prepared at the 39th JECFA (1992), published in FNP 52 Add 1 (1992). Metals and arsenic specifications revised at the 55th JECFA (2000). An ADI of 0-3 mg/kg bw was established
More informationRAFFINOSE/ SUCROSE/ GLUCOSE
www.megazyme.com RAFFINOSE/ SUCROSE/ GLUCOSE ASSAY PROCEDURE K-RAFGL 06/5 (20 Assays per Kit) Megazyme International Ireland 205 INTRODUCTION: Grain legumes are an important component of both human and
More informationSequential Extraction of Plant Metabolites
ISSN: 2319-7706 Volume 4 Number 2 (2015) pp. 33-38 http://www.ijcmas.com Original Research Article Sequential Extraction of Plant Metabolites Shankar L. Laware* PG. Department of Botany, Fergusson College
More informationCORESTA Recommended Method No. 85
Cooperation Centre for Scientific Research Relative to Tobacco Routine Analytical Chemistry Sub-Group CORESTA Recommended Method No. 85 TOBACCO - DETERMINATION OF THE CONTENT OF TOTAL ALKALOIDS AS NICOTINE
More informationExperiment 9. NATURE OF α-amylase ACTIVITY ON STARCH
Experiment 9 NATURE OF α-amylase ACTIVITY ON STARC In Experiment 1 we described the action of α-amylase on starch as that of catalyzing the hydrolysis of α-1,4-glucosidic bonds at random in the interior
More informationCorn Starch Analysis B-47-1 PHOSPHORUS
Corn Starch Analysis B-47-1 PHOSPHORUS PRINCIPLE SCOPE The sample is ignited in the presence of a fixative to destroy organic matter and convert phosphorus to inorganic phosphates which are not volatilized
More information» Croscarmellose Sodium is a cross linked polymer of carboxymethylcellulose sodium.
BRIEFING Croscarmellose Sodium, NF 22 page 2856 and page 702 of PF 30(2) [Mar. Apr. 2004]. A modification is made in the test for Degree of substitution to correct the endpoint color to agree with the
More informationRoti -Quant universal
Roti -Quant universal Colorimetric protein concentration analysis 0120.1 A. The kit contains Roti -Quant universal Reagenz 1 (0118): 500 ml (0120.1) / 200 ml (0120.2) Danger H318-H315 P280- P305+P351+P338-P310
More informationBIOL 305L Spring 2019 Laboratory Six
Please print Full name clearly: BIOL 305L Spring 2019 Laboratory Six Osmosis in potato and carrot samples Introduction Osmosis is the movement of water molecules through a selectively permeable membrane
More informationCoQ10(Coenzyme Q10) ELISA Kit
CoQ10(Coenzyme Q10) ELISA Kit Catalogue No.: EU0196 Size: 48T/96T Reactivity: Universal Detection Range: 0.781-50ng/ml Sensitivity:
More information2. MATERIALS AND METHODS
2. MATERIALS AND METHODS 2.1 Seeds and germination Experiments were carried out in laboratory of Biosciences department of Veer Narmad South Gujarat University, Surat. The seeds of Cajanus cajan and Trigonella
More informationFigure 2. Figure 1. Name: Bio AP Lab Organic Molecules
Name: Bio AP Lab Organic Molecules BACKGROUND: A cell is a living chemistry laboratory in which most functions take the form of interactions between organic molecules. Most organic molecules found in living
More informationARTESUNATE TABLETS: Final text for revision of The International Pharmacopoeia (December 2009) ARTESUNATI COMPRESSI ARTESUNATE TABLETS
December 2009 ARTESUNATE TABLETS: Final text for revision of The International Pharmacopoeia (December 2009) This monograph was adopted at the Forty-fourth WHO Expert Committee on Specifications for Pharmaceutical
More informationHydroxypropyl Starch (Tentative)
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Hydroxypropyl Starch (Tentative) This monograph was also published in: Compendium
More informationEnzyme activity Page 1 of 8
Enzyme activity Page 1 of 8 Contains: - protease - L-glutaminase - Leucine aminopeptidase - sucrase - amylase - cellulase - lipase - catalase - carboxypeptidase Protease Assay Based on: Frankena J., van
More informationKinetics analysis of β-fructofuranosidase enzyme. 1-Effect of Time Incubation On The Rate Of An Enzymatic Reaction
Kinetics analysis of β-fructofuranosidase enzyme 1-Effect of Time Incubation On The Rate Of An Enzymatic Reaction Enzyme kinetics It is the study of the chemical reactions that are catalyzed by enzymes.
More informationHigh-density Lipoprotein Cholesterol (HDL-C) Assay Kit
(FOR RESEARCH USE ONLY. DO NOT USE IT IN CLINICAL DIAGNOSIS!) High-density Lipoprotein Cholesterol (HDL-C) Assay Kit (Double reagents) Catalog No: E-BC-K221 Method: Colorimetric method Specification: 96T
More informationLab #4: Nutrition & Assays for Detecting Biological Molecules - Introduction
Lab #4: Nutrition & Assays for Detecting Biological Molecules - Introduction Most biological molecules fall into one of four varieties: proteins, carbohydrates, lipids and nucleic acids. These are sometimes
More informationNitrate/Nitrite Assay Kit Manual Catalog #
BIOO RESEARCH PRODUCTS Nitrate/Nitrite Assay Kit Manual Catalog # 1305-01 This kit is manufactured to the international quality standard ISO 9001:2008. ISO CI#: SARA-2009-CA-0114-01-B BIOO Scientific Corp.2011
More informationData sheet. TBARS Assay kit. (Colorimetric/Fluorometric) Kit Contents. MDA-TBA Adduct. 2-Thiobarbituric Acid. Cat. No: CA995.
Data sheet Cat. No: CA995 TBARS Assay kit (Colorimetric/Fluorometric) Introduction Oxidative stress in the cellular environment results in the formation of highly reactive and unstable lipid hydroperoxides.
More informationBiological molecules = Biomolecules = Compounds of life
Biological molecules = Biomolecules = Compounds of life Carbohydrates Proteins & Amino Acids Mono-saccharides Olego-saccharides Di-saccharides Poly-saccharides Lipids Oils & Fats Amino acids Proteins Enzymes
More informationA Revised Method for Determining Phosphate-Phosphorus Levels in Sugar Beet Leaf Petioles 1
A Revised Method for Determining Phosphate-Phosphorus Levels in Sugar Beet Leaf Petioles 1 G. E. VARVEL, G. A. PETERSON and F. N. ANDERSON 2 &ceived/orpublicalionj..ne 1,1976 Knowledge of plant nutrient
More informationBRIEFING. Nonharmonized attributes: Identification, Heavy metals, Characters, Labeling, Bacterial endotoxins, Sterility, Storage.
BRIEFING Citric Acid, Anhydrous, page 872 of PF 28(3) [May June 2002]. The European Pharmacopoeia is the coordinating pharmacopeia for the international harmonization of the compendial standards for the
More informationENZYME ACTIVITY. Practical 3
Practical 3 ENZYME ACTIVITY BACKGROUND Enzymes speed up chemical reactions by lowering activation energy (that is, the energy needed for a reaction to begin). In every chemical reaction, the starting materials
More informationGENERAL TESTS FOR CARBOHYDRATE. By Sandip Kanazariya
GENERAL TESTS FOR CARBOHYDRATE By Sandip Kanazariya Introduction Carbohydrates are of great importance to human beings. They are major part of our diet, providing 60-70% of total energy required by the
More informationCOMPARISON AND BIOCHEMICAL ESTIMATION OF THREE PRIMARY METABOLITES OF MEDICINALLY IMPORTANT PLANT AMLA (PHYLLANTHUS EMBLICA)
International Journal of Education and Science Research Review E- ISSN 2348-6457 Volume-1, Issue-3 June- 2014 P- ISSN 2349-1817 COMPARISON AND BIOCHEMICAL ESTIMATION OF THREE PRIMARY METABOLITES OF MEDICINALLY
More informationSALIVA TEST Introduction
SALIVA TEST Introduction This is a practical lesson using saliva to learn digestive enzyme activity. We can check the existence of reducing sugars clearly by Benedict s reaction after salivary enzyme decomposes
More informationGLYCOGEN BEFORE THE LAB YOU HAVE TO READ ABOUT:
GLYCGEN BEFRE THE LAB YU HAVE T READ ABUT:. Glycogen structure. 2. Glycogen synthesis and degradation (reactions with structural formulas and enzymes). 3. The role of glycogen in liver and muscles. INTRDUCTIN
More informationBRIEFING Assay + + +
BRIEFING Sodium Starch Glycolate, NF 22 page 2933 and page 3202 of PF 22(6) [Nov. Dec. 1996]. The United States Pharmacopeia is the coordinating pharmacopeia for the international harmonization of the
More informationAMYLOGLUCOSIDASE from ASPERGILLUS NIGER, var.
AMYLOGLUCOSIDASE from ASPERGILLUS NIGER, var. SYNONYMS INS No. 1100 Prepared at the 59 th JECFA (2002) and published in FNP 52 Add 10 (2002), superseding tentative specifications prepared at the 55 th
More informationAZO-WHEAT ARABINOXYLAN
www.megazyme.com ASSAY OF endo-1,4-b-xylanase using AZO-WHEAT ARABINOXYLAN S-AWAXP S-AWAXL 05/17 Megazyme 2017 PRINCIPLE: This assay procedure is specific for endo-1,4-β-d-xylanase activity. On incubation
More informationExperiment 20 Identification of Some Carbohydrates
Experiment 20 Identification of Some arbohydrates arbohydrates are the direct product of the photosynthetic combination of carbon dioxide and water. By weight, they are the most common organic compounds
More informationHAGEDORN AND JENSEN TO THE DETER- REDUCING SUGARS. MINATION OF LARGER QUANTITIES OF XIV. AN APPLICATION OF THE METHOD OF
XIV. AN APPLICATION OF THE METHOD OF HAGEDORN AND JENSEN TO THE DETER- MINATION OF LARGER QUANTITIES OF REDUCING SUGARS. By CHARLES SAMUEL HANES (Junior Scholar of the Exhibition of 1851). From the Botany
More informationPetrolatum. Stage 4, Revision 1. Petrolatum is a purified semi solid mixture of hydrocarbons obtained from petroleum.
1 001-1208PDG.pdf Petrolatum Stage 4, Revision 1 Definition Petrolatum is a purified semi solid mixture of hydrocarbons obtained from petroleum. It may contain a suitable antioxidant. Description and Solubility
More informationFood acidity FIRST LAB
Food acidity FIRST LAB objective To determine total acidity of milk, juice, vinegar and oil acid value Food acidity Food acids are usually organic acids, with citric, malic, lactic, tartaric, and acetic
More informationASSAY OF using AZO-FRUCTAN S-AZFR5 11/17
www.megazyme.com ASSAY OF endo-fructanase using AZO-FRUCTAN S-AZFR5 11/17 Megazyme 2017 PRINCIPLE: The substrate is the high molecular weight fraction of chicory fructan (DP ~ 20-60) dyed with an azo-dye
More informationSMOKE FLAVOURINGS. Wood smoke flavour, Smoke condensate
SMOKE FLAVOURINGS Prepared at the 57th JECFA (2001) and published in FNP 52 Add. 9 (2001), superseding tentative specifications prepared at the 55th JECFA (2000) and published in FNP 52 Add. 8 (2000).
More informationFeedstuffs Analysis G-22-1 PROTEIN
Feedstuffs Analysis G-22-1 PROTEIN PRINCIPLE SCOPE Many modifications of the Kjeldahl method have been accepted for the estimation of protein in organic materials. It comprises sample oxidation and conversion
More information» Monohydrate Citric Acid contains one molecule of water of hydration. It contains not less than 99.5 percent and not more than 100.
BRIEFING Citric Acid, Monohydrate. The European Pharmacopoeia is the coordinating pharmacopeia for the international harmonization of the compendial standards for the Citric Acid, Monohydrate monograph,
More information6.02 Uniformity of Dosage Units
6.02 Uniformity of Dosage Units Change 1. Content Uniformity, 3. Criteria and Table 6.02-2 as follows: 1. Content Uniformity Select not less than 30 units, and proceed as follows for the dosage form designated.
More informationAppendix II. Barton's reagent:
Appendix II SOLUTIONS AND REAGENTS Barton's reagent: A. Dissolved 25 g ammonium molybdate in 400 ml de-ionized water (15 ). B. Dissolved 1.25 g ammonium metavanadate in 300 ml of boiling de-ionized water
More information--> Buy True-PDF --> Auto-delivered in 0~10 minutes. GB Translated English of Chinese Standard: GB5009.
Translated English of Chinese Standard: GB5009.259-2016 www.chinesestandard.net Sales@ChineseStandard.net NATIONAL STANDARD GB OF THE PEOPLE S REPUBLIC OF CHINA National food safety standard Determination
More informationTitle Revision n date
A. THIN LAYER CHROMATOGRAPHIC TECHNIQUE (TLC) 1. SCOPE The method describes the identification of hydrocortisone acetate, dexamethasone, betamethasone, betamethasone 17-valerate and triamcinolone acetonide
More informationThis chapter deals with the evaluation of alpha amylase inhibitory
This chapter deals with the evaluation of alpha amylase inhibitory activity of different extracts isolated from leaves of Aloe vera L. and leaves of Azadiracta indica A Juss. collected from Bharatpur and
More informationPectins. Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Pectins This monograph was also published in: Compendium of Food Additive Specifications.
More informationEnzyme Action: Testing Catalase Activity
Enzyme Action: Testing Catalase Activity Pennsylvania Science Standards: S11.A.1.1.4 S11.A.1.3.1 S11.A.2.2.2.1 S11.A.2.2.2.2 Keystone Eligible Content Bio.B.4.1.1, Bio.B.4.1.2, and Bio.B.4.2.5 Introduction
More informationOCR (A) Biology A-level
OCR (A) Biology A-level Topic 2.2: Biological molecules Notes Water Water is a very important molecule which is a major component of cells, for instance: Water is a polar molecule due to uneven distribution
More informationDate... Name... Group... Urine sample (Tube No 2)
Date... Name... Group... Instructions for the practical lesson on biochemistry Topic: Non-protein nitrogen compounds Task 1: Estimation of creatinine in serum and urine 1. Trichloroacetic acid 1.22 mol/l
More informationDetermination of Tanninoids. Analytical Pharmacognosy
QUIZ If the manager of a phytopharmaceutical industry wish to buy one chromatographic equipment, which one you recommend, HPLC or TLC densitometer. What are the reasons to support your recommendation.
More informationNitrate and Nitrite Key Words: 1. Introduction 1.1. Nature, Mechanism of Action, and Biological Effects (Fig. 1)
7 Nitrate and Nitrite Key Words: Nitrate; nitrite; methemoglobin; blood pressure; asphyxia; spinach; spongy cadmium column; zinc metal; sodium nitrate; sodium nitrite; ammonia buffer solution; Jones reductor.
More informationCatalase Lab - A Bio ENZYME ACTIVITY Investigation Created by Gen Nelson, modified by Dr G
Catalase Lab - A Bio ENZYME ACTIVITY Investigation Created by Gen Nelson, modified by Dr G INTRODUCTION Hydrogen peroxide (H 2O 2) is a poisonous byproduct of metabolism that can damage cells if it is
More informationQUALITATIVE TESTS OF CARBOHYDRATE
QUALITATIVE TESTS OF CARBOHYDRATE MACROMOLECULE CARBOHYDRATES Are the key source of energy used by living things. Also serve as extracellular structural elements as in cell wall of bacteria and plant.
More informationBCH302 [Practical] 1
BCH302 [Practical] 1 Carbohydrates are defined as the polyhydroxy aldehydes or polyhydroxy ketones. Most, but not all carbohydrate have a formula (CH 2 O)n (hence the name hydrate of carbon). Sugars ends
More information