Author: LAW JIA JUIN
Content of this report:
1. INTRODUCTION
2. FORMULA OF OUR PREPARATIONS
3. CALCULATIONS BEFORE PREPARATION
4. RESULTS OF PRODUCT EXAMINATION AND EVALUATION
5. DISCUSSION
6. CONCLUSION
7. REFERENCES USED.
1. INTRODUCTION
This is an article written by me which mainly discuss on the preparation of sterile injection .An injection is a method in which fluid is put into body by the method of infusion, this is usually done via a hollow needle and a syringe which will pierced through the skin to a sufficient depth for the material to be forced into the body. An injection can be of intravenous type, intramuscular type or subcutaneous type. All injections share some common characteristics which are good sterility, free from microorganisms, free of external impurities and particles, free of pyrogens as well as endotoxin (or consist only the minimum acceptable amount)and should be kept in a well -sealed glass container to prevent contamination or leakage. .Since these properties are important for the patient and have a direct effect on the tolerance of patient towards the injection, thus they are usually tested after the preparation of injections. The injection products are tested and evaluated from four aspects which are, sterility of product, clarity of product, leakage testing, and last but not least the endotoxin testing. In our practical ,we tested the leakage test , then follow by the clarity test, third sterility test, and lastly we tested the endotoxic test. The purposes and methods of these evaluations will be discussed later by me. The calculations we used before the preparations are also given by me.
2. FORMULA OF OUR PREPARATIONS
Dextrose injection B.P
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Anhydrous dextrose
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5.0g
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Water for injection to
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100ml
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Lignocaine HCL Injection B.P
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Lignocaine HCL
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2.0g
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Sodium chloride
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q.s
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Water for injection to
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100ml
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Lactated Ringer’s injection B.P
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Sodium chloride
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600mg
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Sodium lactate
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320mg
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Potassium Chloride
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40mg
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Calcium Chloride
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27mg
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Water for injection to
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100ml
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3.CALCULATION WE DID BEFORE PREPARING OUR RESPECTIVE INJECTIONS SOLUTIONS
1. Concentration of anhydrous dextrose
W=0.3M/N
M=180 N=1 (because dextrose is an organic substances with no charge)
W=0.3x 180 =54g/l =5.4g/100ml= 5.4%w/v BP specification =5%w/v
Therefore w is acceptable. And 5.4g of anhydrous dextrose powder should be dissolved in sterile water and made up to 100ml solution using a volumetric flask.
2.Concentration of NaCl required to make lignocaine HCl injection isometric with blood plasma.
[0.52- (a x % of drug solution)] / b
a= Freezing point of 1 % w/v lignocaine =0.13 o C
b= Freezing point of 1 % w/v NaCl= -0.576
[0.52-(0.13 x 2)] / (0.576)
=45mg/100ml
45 mg of NaCl solids should be weighed, dissolved and made up to 100ml of lignocaine HCl solution, making the lignocaine HCl solution isometric to human blood plasma.
3. Combined effective molar concentration (EMC ) of ringer lactated solution.
1.NaCl
NaCl= 600mg/100ml= 6g/l
W=(C x M) /N W=6g/l M=58.2 g N=2
C= (6 x 2)/ 58.5= 0.205 M ……….(A)
2.Lactate
Solution lactate =320mg/100ml= 3.2g/l, thus W =3.2g/l
W=(C x M) /N
3.2= (C x 112.07)/2
C=0.05710 M …………(B)
Using the same formula ,W=(C x M) /N , and the values of M and C provided we can actually calculate for the C value of KCl and CaCl2:
3, KCl= 0.01073M………(C)
4, CaCl2= 0.00723M……..(D)
EMC= A+ B+ C+ D =0.28018 =0.3 M
Concentartion of NaCl for iso-osmotic solution
= W=(C x M) /N M=58.5 which is the molecular weight of NaCl ;N=2 (charge of Na plus charge of Cl)
=[0.3 x 58.5]/ 2
= 8.775g/l =0.8775g/100ml =0.9g/100ml= 0.9%w/v
4. RESULTS WE GET AFTER EVALUATING OUR GROUP’S PRODUCT, DEXTROSE INJECTION BP
A) CLARITY TEST
OBSERVATION: The injection solution we prepared last week is clear and free from external contaminants and particles when observed under the light with both black and white backgrounds.
LEAKAGE TEST
B)
B)
OBSERVATION: There is no red dye moving into our injection solution, showing us that the glass container holding our injection solution is well sealed.
C) STERILTY TESTING
OBSERVATIONS:
TEST TUBE 1 (nutrient medium with product): no turbidity showing that there is no growth of microorganism the injection product is well sterile
TEST TUBE 2 (nutrient medium with with product): no turbidity showing that there is no growth of microorganism the injection product is well sterile.
CONTROL TUBE (nutrient medium with without product): no turbidity
D) BACTERIAL ENDOTOXIN TEST.
OBSERVATIONS:
PPC TUBE: Clot is formed
SPL TUBE: No clot formed
5. DISCUSSION
Right after the injection solution is produced and packed inside a glass container which is then sealed, we checked the clarity of our injection solution, this is because every strict and good manufacturing practice requires a visual inspection of each ampoule individually, this is usually done with our naked eyes. If any visible particles are spotted in our container, we should actually discard the product, this is because contamination is said to be occurred during the process of manufacturing. This inspection should be done with human inspectors under a condition of good light source, and is usually done against a black and white background. In our examination we found out that the dextrose solution we prepared is totally clear of any particles. The solution is transparent and clear, showing that our product had passed the clarity test.
In the leakage test, the sealed-glass container which holding our dextrose solution is put inside a permeable bag with marbles inside it, then the whole bag is immersed into a basin containing red dyes. The permeable bag containing the injection sealed glass container is allow to be immersed in the dye for 5 minutes, after that it is taken out for examination. If let say, the transparent injection solution in the sealed container is contaminated with the red dye, then our product is said to be failed from leak test, and it had to be discarded, an improper sealed container is unacceptable because this can allow the outer air, microorganisms, dusts, and contaminants to get into our container, contaminating our injection solution, making it harmful and unacceptable to our patient. Beside that improper sealed container also allow our injection solution to leak out, changing the desired dose for injection. Since there is no red dye found inside our injection product ,we can deduce that our container is well sealed .The injection solution is well kept and protected from contamination, the product passed this test.
The third test we did for our dextrose solution quality examination is sterility test. Before conducting this test, the neck of sealed glass container was craved with knife for easier gripping of the white cap breaker, the top sealed neck of glass container then was broken by using the white cap breaker stated. Tube-dilution method is applied in this test and the medium we used was thioglycollate medium. Two drops of our injection solution was transferred to two test tubes each with a suitable medium in which it provides optimum conditions for bacterial growth and incubate at a suitable temperature (37oC) for sufficient time (24 hours). The transfers of sample solution are done under an aseptic condition inside a turbulence air flow chambers ,this is to ensure an accurate result and to prevent false positive result. A control is also prepared with medium but no injection solution inside. If the medium turns to become turbid then we can deduce that our injection product is contaminated or susceptible to bacterial growth. If there is no turbidity in the medium the preparation may be sterile. Since there are no turbidity observed in all the 3 test tubes, we can say that our injection solution is free from the contamination of microorganisms and again we can conclude that our product passed this sterility test.
In endotoxin test, 0.5ml of injection solution was transferred into a sample tube (SPL tube) using pipette, then we gently swirled the SPL tube for proper mixture of the content. After that we used the same pipette to transfer 0.25ml of solution from the SPL tube to positive product control tube(PPC tube). Upon proper transfer we swirled the PPC tube gently and immediately placed both the tubes into an incubator with temperature 37 degree celcius for 1 day. If both tubes clot, then we will had rejected the product because the injection solution is said to contain more than 0.25 EU/ml of endotoxin. If the control PPC tube did not clot then we will had repeated the test because the result is invalid. But in in our case ,the clot formed in PPC tube but did not formed in SPL tube ,thus we concluded that the endotoxin level in our injection solution is less that 0.25EU/ml, we is acceptable, our product passed this test.
6.Conclusion
: Our product which is the ,dextrose injection solution BP, passed all the 4 tests ( clarity, leakage, sterility and endotoxin test). We concluded that this is a well prepared injection solution with proper sterility and perfect seal as well as ideal packing.
7.References
1. Wikipedia.com.my
2. Elearning.imu.edu.my
3. Pharmaceutics-info.com.my
