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. Ⅰ. Phthalamine (O-T)Basic Principles The aldehyde base ofglucose is synthesized in a solution of hot acetic acid, and then dehydrated to produce Schiff base, which is then rearmed by molecules to produce a set of blue-green compounds with an absorption peak at 630nm, the amount of light absorption is positively in line with the concentration of blood sugar.
equipment is . Water bath tank,
test tube
, hydrometer.
Reagents
Reagents
1.1. Phthalates reagents take ice acetic acid (AR) 940 ml, add thiophenyl (AR) 1.5g, dissolve and add phthalates 60 ml mix well, placed in brown bottles, room temperature preservation. New reagents should be placed for 24 hours (to be aged) for use. This reagent is highly corrosive and should avoid burns caused by contact with the skin.
2. 12mmol/L benzoic acid solution called benzoic acid 1.4g, add distilled water 900ml,
heating
to help dissolve, cooled and distilled water to 1000ml.
3. Glucose standard storage fluid (100mmol/L) accurately called waterless glucose (pre-positioned in an oven at 80 degrees C
drying
to constant weight, stored in a shift dryer) 1.802g Dissolve with 12mmol/L benzoic acid solution, move to a 100ml capacity bottle, dilute to scale with 12mmol/L benzoic acid solution, shake well, move into a brown bottle and store in the refrigerator.
4. The glucose standard application fluid (5mmol/L) accurately absorbs 5.0 ml of the glucose standard storage fluid, places it in a 100 ml capacity bottle, and dilutes it to the scale with a 12mmol/L benzoic acid solution.
the procedure is . Mix well, bring to the boil in a boiling water bath for 12 minutes, remove the running water and cool for 5 minutes with a hydrometer wavelength of 630nm, zero in the blank, read each tube absorbent.
's the . Note that the
. 1. The reaction of phthalates and glucose is not specific, other sugars in the reaction can also produce a similar absorption of glucose
spectral
, their relative absorption ratio is: glucose s 1.00; However, only glucose, fructose and semi-lactose were present in normal people
sery
, and the longer two had very little in the blood of normal people and did not affect the reagent test results.
2. The color intensity of glucose and phthalates is related to reaction conditions (lot number of phthalates and ice acetic acid, time saved after preparation of reagents, and heating temperature and time). Therefore, the reaction conditions of the tubes must be exactly the same.
3. Phthalamine is a light yellow oily liquid, easily oxidized to red-brown, with this phthalates preparation reagents, blank tube color is dark, low sensitivity, seriously affecting the results of the determination. Must be treated by the following methods to apply: in 500 ml of phthalates add hydroxyamine hydrochloride 0.5g, put 50 to 60 degrees C water bath for 20 minutes from time to time shake, so that phthalates reduce, light color. If the above treatment is not ideal, then phthalates are re-distilled with a full glass distiller, collect 199 to 201 degrees C colorless or light yellow outflow liquid, brown bottles and black paper bags to avoid light seal preservation.
II. The effects of insulin and epinephrine on blood glucose . Basic Principles: Inand animals, blood sugar concentrations are
by
hormones and constantly regulated. Insulin lowers blood sugar, while many other hormones have the effect of raising blood sugar, with epinephrine acting more quickly and significantly. Insulin promotes the liver and muscles to synthesize glucose into sugar, and also enhances the oxidation of sugar, so it can lower blood sugar;
the study looked at changes in blood sugar levels before and after the rabbit injected insulin and epinephrine.
equipment:
syringes, needles, blades, tracheometers.
Reagents:
1. Adrenaline and insulin
2. Sodium aureate
3. 25% Glucose
Basic Operations
1. Animals are
take normal rabbits, pre-experiment hunger, weighing (usually 2 to 3 kg).
2. Blood collection
Generally to the ear veins to take blood, first hair, with xylene wipe rabbit ears, so that its blood vessels filled with blood, and then dry with dry cotton balls, in the bleeding site to apply a thin layer of Vyslin, and then with a crude needle punctured veins blood, blood income anticoagulant tube (about 2 mg of sodium per ml of blood) side collection side shake, to prevent coagulation. Press the blood vessels with dry cotton balls to stop the bleeding.
3. After taking the hormone
the blood of the hungry rabbit, one of the rabbits injected insulin under the skin at a dose of 0.75 units/kg body weight and recording the injection time. Take the blood in an hour. Immediately after taking the blood, 25% glucose fluid was injected into the abdominal cavity or under the skin to prevent the rabbit from dying of insulin shock.
injected with epinephrine under the skin of another rabbit at a dose of 0.4 mg/kg body weight and recording the injection time. Take the blood in half an hour.
calculated
rate of decreased blood sugar after insulin injection and increased blood sugar after epinephrine injection.
commonly used chemical methods for blood sugar measurement have phthalates, commonly used enzyme method for glucose oxidase method,
sugar
method. .