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    Home > Chemicals Industry > Chemical Technology > Safety inspection of traditional Chinese medicine injections (5)

    Safety inspection of traditional Chinese medicine injections (5)

    • Last Update: 2022-06-24
    • Source: Internet
    • Author: User
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    (5) Antihypertensive Substance Inspection Law

    1.
    Brief description of this method is to compare the degree of blood pressure drop caused by a certain amount of histamine reference substance and the test substance to determine whether the content of the antihypertensive substance in the test substance meets the prescribed inspection methods

    .

    2.
    Experimental materials and utensils

    (1) Blood pressure recording device: mercury sphygmomanometer, pressure sensor, multi-channel physiological recorder or physiological data recording system
    .

    (2) Experimental equipment: syringe (1ml precision 0.
    01ml, 5ml precision 0.
    2ml), straws, pipettes, volumetric flasks, vials with stoppers, three-way switch, absorbent cotton, thread, rope

    .

    (3) Surgical instruments: operating table, shears, surgical scissors, ophthalmic forceps, hemostatic forceps, scalpel, tracheal intubation, arterial clamp, arteriovenous intubation
    .

    (4) The accuracy of the balance: 0.
    01mg or 0.
    1mg (for weighing the reference substance or test substance), 1mg (for reagent weighing), 100g (for animal weighing)

    .

    (5) Reagents: sodium chloride injection, phenobarbital sodium, pentobarbital sodium , heparin sodium injection and other reagents
    .

    3.
    The experimental animals are healthy and unharmed.
    Domestic cats weighing more than 2kg can be both male and female, and the females are not pregnant

    .

    4.
    Solution preparation

    (1) 10% phenobarbital sodium solution: Weigh an appropriate amount of phenobarbital sodium, add water to make a 10% solution; or use 10% phenobarbital sodium injection
    .

    (2) 1% sodium pentobarbital solution: Weigh an appropriate amount of gobarbital sodium and add water to make a 1% solution
    .

    (3) Heparin sodium solution: Heparin sodium injection is appropriately diluted with sodium chloride injection to prepare a 1000U/ml solution
    .

    (4) Histamine reference substance solution: Take histamine phosphate reference substance and place it at room temperature
    .

    1) Cut the small tube of the reference substance, (be careful not to transfer glass shavings) and accurately weigh an appropriate amount of histamine phosphate
    .

    2) Multiply the weighed histamine phosphate by the coefficient (calculated according to the instructions provided, generally 0.
    342) to convert the actual weight of histamine

    .

    3) Add water to dissolve histamine phosphate to prepare a solution of 1.
    0mg/ml calculated as histamine.
    Dispense it and store it at 4~8℃ for later use

    .
    If there is no precipitation, it can be used within 3 months .

    4) On the day of the experiment, take out the reference solution and place it at room temperature
    .

    5) Precisely measure an appropriate amount of 1.
    0mg/ml reference solution, prepare a 10ug/ml diluent with physiological sodium chloride injection, and then dilute it into 0.
    5, 1.
    0, 1.
    5ug/ml histamine solution, according to the animal The body weight is administered at 0.
    1ml/kg (dose is 0.
    05, 0.
    10, 0.
    15ug/kg)

    .
    The volume of administration can be adjusted under special circumstances, but the amount of histamine given to animals per kg body weight cannot be changed

    .

    (5) Test solution: According to the dosage specified under each category, make the test solution of appropriate concentration
    .
    During the experiment, it is required that the injection volume of the test solution and the reference substance diluent is equal

    .

    5.
    Animal anesthesia and surgery

    (1) Anesthesia and fixation: Weigh the animal; draw 10% phenobarbital sodium solution (1ml/kg) and 1% pentobarbital sodium solution (1ml/kg) with a syringe, mix well, and anaesthetize the animal by intraperitoneal injection , You can also use other anesthetics to anesthetize the animal; after the animal is anesthetized, fix the animal on the operating table supine with a rope, cut the hair on the neck and thighs with shears, and try to keep the animal's body temperature
    .

    (2) Surgery and arteriovenous cannulation:

    1) Make an incision along the midline of the neck, carefully separate the trachea, perform tracheal intubation, and connect an artificial ventilator if necessary
    .

    2) Separate one side of the common carotid artery, peel off the attached fat tissue and nerves, and thread 2 threads under the artery, one is ligated to the distal end of the artery, and the proximal end of the artery is clamped with an arterial clamp
    .

    3) Separate the femoral vein and perform venous cannulation
    .

    6.
    Adjustment of pressure measuring device and recorder (according to the operation method of pressure sensor and recorder)

    (1) Connect the mercury sphygmomanometer, arterial cannula, and pressure sensor to the recording instrument or recording system
    .

    (2) Use normal saline to exhaust the air in the pressure sensor, mercury sphygmomanometer and arterial cannula (each connection must be firm and leak-proof, such as water leakage will affect the blood pressure measurement)
    .

    (3) Turn on the power of the recorder and turn on the recorder or recording system
    .

    (4) Use physiological saline to pressurize, calibrate with mercury sphygmomanometer or calibrate the pressure sensor, perform zero adjustment and baseline calibration, etc.
    , to ensure the accuracy of blood pressure measurement

    .

    (5) Cut a small opening in the carotid artery, insert the arterial cannula (or use a 12-gauge short blunt needle) and ligate the cannula and artery with another thread, so that the cannula and artery are in a natural state and avoid twisting the artery.
    Affect the measurement of blood pressure

    .

    (6) Open the arterial clamp and inject about 0.
    5ml of
    1000u/ml
    heparin solution into the tee on the cannula to prevent blood clotting and block the cannula from affecting blood pressure measurement
    .

    (7) Use normal saline to exhaust the air in the venous cannula, insert a needle into the venous cannula (or use a scalp needle) in the femoral vein, fix the cannula, and inject an appropriate amount of heparin solution for anticoagulation
    .

    (8) After the operation is completed, use a little absorbent cotton dipped in saline and cover the arterial and venous cannula
    .

    (9) Measurement of normal blood pressure Turn on the recorder or recording system to measure and record normal blood pressure
    .

    7.
    Determination of animal sensitivity

    (1) Adjust the recorder or recording system to an appropriate speed, inject the reference substance dilution 0.
    5ug/ml, 0.
    1m/kg from the intravenous cannula according to the animal's body weight, immediately inject appropriate volume of saline, and flush the liquid medicine into the body , Record the blood pressure drop curve

    .

    (2) When the blood pressure rises back to the baseline, inject the second reference diluent 1.
    0ug/ml, 0.
    1ml/kg, immediately inject an appropriate volume of normal saline, flush the liquid medicine into the body, and record the blood pressure drop curve

    .

    (3) After the blood pressure returns to the normal value, inject the third reference diluent 1.
    5ug/ml, 0.
    1ml/kg, immediately inject an appropriate volume of normal saline, flush the liquid medicine into the body, and record the blood pressure drop curve

    .

    (4) Repeat the administration 2 to 3 times
    .

    (5) If the blood pressure drop caused by the dose of 0.
    10ug/kg is not less than 2.
    67kPa (20mmHg), and the average value of the response caused by each dose is different, it can be considered that the sensitivity of the animal meets the requirements

    .

    (6) When measuring sensitivity and dosing, the interval between two adjacent dosings is the same (3 to 5 minutes).
    Each dosing should be performed after the previous response has stabilized

    .

    8.
    Administration (in the order of ds, dT, dT, ds)

    (1) According to the 0.
    10ug/kg reference substance diluent dosage (ds), inject into the animal through an intravenous cannula, and immediately inject an appropriate volume of normal saline into the body, and record the blood pressure drop curve

    .

    (2) The test product shall be administered at the prescribed dose (dT) under the pharmacopoeia category, the method is the same as ds, and the blood pressure curve shall be recorded after administration
    .

    (3) Do the second administration of dT as above, and record the blood pressure drop curve
    .

    (4) Do the second dose of ds as above, and record the blood pressure drop curve
    .

    9.
    Result Judgment

    (1) Measure the magnitude of blood pressure reduction in mmHg for each dose or convert it into kPa units
    .

    (2) Compare the reactions caused by the first and third, second and fourth doses respectively
    .

    (3) If the response value caused by dT is not more than half of the response value caused by ds, it is considered that the antihypertensive substance inspection of the test product meets the requirements, otherwise, continue to inject 1 group of 4 doses in the above order, and follow the same method Compare the response caused by the ds and dT doses in the two groups respectively.
    If the response value caused by dT is not greater than the response value caused by ds, it is still considered that the antihypertensive substance inspection of the test product meets the requirements

    .

    (4) If the response value caused by dT is greater than the response value caused by ds, it is considered that the antihypertensive substance inspection of the test product does not meet the requirements
    .
    Otherwise, another animal should be retested.
    If the retest results still have a response value caused by dT that is greater than the response value caused by ds, it is considered that the antihypertensive substance inspection of the test product does not meet the requirements

    .

    10.
    Matters needing attention

    (1) If it is necessary to measure multiple samples on the same animal, the sensitivity should be checked again.
    If it still meets the requirements, the experiment can be carried out, and so on

    .

    (2) Animal anesthesia can be anesthetized by intraperitoneal injection of pentobarbital sodium solution 40~55mg/m/kg alone according to laboratory experience, or other suitable anesthetics can be used for anesthesia
    .

    (3) During the experiment, the animal can be kept warm with a constant temperature operating table or illuminated by an operating lamp to stabilize the animal's blood pressure
    .

    (4) If the animal’s trachea is incised, pay attention to removing blood clots in the trachea (usually with absorbent cotton) to prevent the animal from breathing difficulties and affecting the stability of blood pressure
    .

     

     

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