aT quate ANALYSIS Naregen Osten ty peat ig ttowen fn os sate ssh as ne te cannot be used for nitrogen estimation by Kjeldshl's method, However, the use af geese dung digestion reduces miagen bah pete ever eee vii adele thal Cee ea mre (i) The tiation may be ear: indicctor (the first the absorveat, 1 out by 0.02 (N) HCH solution with methyl redanethylene blue mixed srance of violet colour indie hi etl pint) in case hotie acid solution is used as ii) A blank test should be performed wit hes pe tine of the original Se E analytical glucose and the titte value of the origi: (iv) 0.02 (N) NaOH may be used for tiation of eveess acid using phenolphatateia as indicator nd saturated 0.02 (N) oxalic acid solution have to be used for determination of strengtty of sodium hydroxide solution. : 2. To determine the strength of iple + A known volume of Febiling's solution is ceduced 69 ed unknown strengths. ‘The st the volume of the sugar sol piven glucose solution using Kebling's solu ric oxide by glucase solution of «supplied ghicose solution in is then eateutated (rom on consumed, Cat 20o + 2Cuh (tartarate complerd. Glucose Feinling’s soln, Red ppt Glovonic acia Materials required and th Fehling > Cu,0 4 Clty 207 A: 17.32 ain, of hydsated copper sulphate is issolved in 250 ml, of distilled water, inne Unt fe gad 25 gm. of sodium hydroxide is dissolved in 250 Standard glucose s weighing bout! Methylene blue indicator £1.25 ym. of pure wah} Jacose (A + R) is weighed accurately from a ng volume is made up to the math, = DSW vere & : 0.5 gi, of methytone blue dissolved in 250 mi, of distilled water. ied [Appeon 8 Unknown solution + Will Procedure : Pelli Fehiing B in a clean dry bottle or cei onied! flask 25 mil, of distilled water is added, boiles over a wite soit eran a burette, first i T mil. postion aati he solution assures drop wail the blue colour just disappears keepin hs im stare boii ion. This gives. gm approxienate value, Addition of metlytene rulicator just before the end point (only 0.5-- Unk. of glucose solution is left for reaching end point) sharpens end point, ‘The process is repeated until a sencordant result has been reached, Similar titrations ate performed at least three times with supplied coution when concordant readings we expected Experimental Result + Table 1. Weiehing. of glucose for standard solution of 250 ml > gm. dissolved in UHL] solution is prepared by thorough mixing of equal volumes of Febli flask, 25 ml of itis pipetted out for each titeat A and into a 250 mb ieose solution is faint blue colour and then drop by wre and Tie weigh [ Final wt wy pm wp gm, Table Li, Stands ation of Kefiling's solution + Vol. of Behl soln. | Vol, of glucose solution | Mean volume of glucose] required tn,nation of strength of unknown Same as Table U. ulation : Weight of glucose dissolved in 250 ml, = w gm. Vol. of Fehliny 25 ml. Mean volume of st Vk, Mean volume of supplied (unknown) glucose solution = Vu. “olution taken for cach titratio ndard (known) glucose solution 250 ml. of standard glucose solution = w gm, glucose. Vk mot " SAVE gm, glucose, F wx Vk Again Vu of unknown glucose solution == Vk ml. of standard glucose solution = gm. ¢ ul 250 nlucose. x Vk x 4 2 1,000 ml. of unknown glucose solution weve x 1.000 =a Bm. of glucose. c. Strength of unknown glucose solution 2 VEX4 gmJflitre. N.B. : Percentage of purity of glucose sample may be checked by preparing both the glucose solution standard one using A.R, grade anhydrous glucose and the other from given glucose sample. Titrations ar performed using same Fehiling’s solution for each titrations. ‘The working formula may be expressed as % of glucose =vExw x 100 ple dissolved in 250 ml. of distilled water and w, Vk, Vu are of usuz Where s = weight of glucose s meaning in the above experiment, 3. Determination of strength of Sucrose solution by Fehling's solution, Principle : The non-reducing sugar sucrose is estimated by converting it into a mixture of tw: reducing sugars, namely glucose and fructose by boiling with dilute acids. CypH 9011 + HzO —— CoH20g + CeH12.. sucrose glucose fructose i.e. 342 gm, of sucrose = 360 gm. of ‘invert sugars’ ‘The ‘invert sugar’ so produced is titrated by standard Fehling’s solution using methylene blue a indicator. Materials required and their preparation : Will be supplied. (approx 4.5 gm. dissolved in 1 litre). Unknown Solution Preparation of standard glucose solution, Fehling's solution etc. are performed as described unde estimation of glucose. Procedure : Standardisation of Fehling solution : As described under estimation of glucose. n of unknown sucrose solution with standard Fehling's solution : Estimat 25 ml. of given (supplied) sucrose solution is pipetted out into a 250 ml. conical flask and boiled witk 25 mi of A(N) HCI [8 ml, of 12(N) HCI diluted 10 25 ml. with water] at 60° - 70°C for 20 minutes on a 2 bath, The solution is cooled and neutralised with 30% sodium hydroxide solution using methyl red a8 weer ba ‘The resulting solution is transfered quantitatively to 2250 ml. volumetri flask; volume is made into the mark with distilled water andthe solution is irated with standard Fehling's solution following the procedure given for estimation of glucose.