Quality control and evaluation of parenteral products:

Three general areas of quality control are:

Raw material / incoming stock control

Manufacturing / in-process control

Finished product control

Incoming stock control:

routine tests on all ingredients

pyrogen tests on WFI

glass tests on glass containers

tests on rubber closures

Microbial load (bioburden) tests etc.

Manufacturing / in-process control:

Conductivity measurements during the distillation of WFI

Confirmation of volume of fill in product containers

Monitoring of time and temperature for thermal sterilization of the product etc.

Finished product control:

Final assays and other tests like pH, tonicity, preservative concentration

Sterility test

Clarity test/ Particulate matter

Leakage test

Pyrogen test

Final assays and other tests like pH, tonicity, preservative concentration: These tests are done as per product requirement and specification/s.

2) Sterilitytests:- The methods which are used to perform sterility tests are

a) Directtransfermethod.

b) Membranefiltration method.

A) Direct Transfer method: it is a traditional sterility test method which involves a direct inoculation of required volume of a sample in two test tubes containing a culture medium [fluid thioglycollate media (FTM), Soya-bean casein digest (SCDM)].

B) Membrane Filtration method:This method basically involves filtration of sample through membrane filters of porosity 0.45 micron and diameter about 50 mm. The filtration is assisted under vacuum or pressure difference. After filtration, the membrane is washed with a sterile diluting fluid to remove traces of bacteriostatic agents etc. and aseptically removed. The membrane is cut into 2 halves One-half of the membrane is placed in a suitable volume (usually 100 ml) of FTM, and the other membrane half in SCDM.

INCUBATION-PERIOD

All test containers should be incubated at temperatures specified by the pharmacopoeia for at least 14 days, regardless of whether filtration or direct inoculation test methodology is used.

*Interpretation: If no visible evidence of microbial growth in culture medium in test tubes is found, then it is interpreted that the sample representing the lot is sterile.

Appropriate positive and negative controls must be included in the sterility testing experiments

SAMPLING

The number of containers tested per batch and quantity tested from each container should be in accordance with the pharmacopoeial specification.

3) Particulatemattertesting:- Particulate matter is of primary concern in the parenteral products given by I.V. Route. All parenteral products should be free from insoluble particle.

Biological risk associated with particulatematter:-Inflammatory response-Antigenic response-Occlusion of blood vesselsFurther U.S.P. states that GMP Requires that all containers be visually inspected and that with visible particle be discarded.

Methods for monitoring particulate matter contamination:

Visual method

Coulter counter method

Filtration method

Light blockage method

4) Leaker Test: - The leaker test is intended to detect incompletely sealed ampoules, so that they may be discarded. This test is usually performed by producing a negative pressure within an incompletely sealed ampoule, while the ampoule is submerged entirely in a deeply colored dye solution. Most often, approximately 1% methylene blue solution is employed. After carefully rinsing the dye solution from the outside, color from the dye will be visible within a leaker. Vials and bottles are not subjected to such a leaker test, because the sealing material (rubber stopper) is not rigid.

5. Pyrogen Test: - pyrogens are lipopolysacchrides chemically and heat stable and are capable of passing through bacteria retentive filter.

When these pyrogens are introduced into a body they produce a mark response of fever with body ache and vasoconstriction within an onset of 1 hour.

The USP evaluates the presence of pyrogens in parenteral preparations by a qualitative fever response test in rabbits, the Pyrogen Test, and by the Bacterial Endotoxins Test LAL Test.

Fever response test in rabbits Why Rabbit?

Reproducible pyrogenic response

Other species not predictable

Similar threshold pyrogenic response to humans

For the test:

Rabbits must be healthy and mature

New Zealand or Belgian Whites used

Either sex may be used

Must be individually housed between 20 and 23C

Preliminary test (Sham Test)

Intravenous injection of sterile pyrogen-free saline solution

To exclude any animal showing an unusual response to the trauma (shock) of injection

Any animal showing a temperature variation greater than 0.5 C is not used in the main test

All glassware, syringes and needles must be pyrogen free.

Main test:

Group of 3 rabbitsPreparation and injection of the product:-Warming the product to 372C-Dissolving or dilution-Injection site: ear vein-The injected volume: about 10 ml per kg of body weight over 10 min. duration-Record temperature at 30-min intervals for 3 hours.-Site of temperature measurement- rectal Instrument-thermometers/ thermocouples

Interpretation of the results:

The sample may be judged nonpyrogenic if no single rabbit shows a rise in temperature of 0.5C or greater above its control temperature.

If this condition is not met, the test must proceed to a second stage. In the second stage, five additional rabbits are given a new preparation of the same test sample as the original three rabbits.

The solution may be judged non-pyrogenic if not more than three of the eight rabbits show individual temperature rises of 0.5C or more.

Disadvantages:

High variability in response.

Difficulty in controlling all factors.

Antipyretic drugs such as aspirin, acetaminophen and morphine mask pyrogenic effect (i.e.,misleading results).

Some other drugs have their inherent pyrogenic effect.

LAL test:-

The test was developed in 1960s by Drs. Bang and Levin and is based on clotting rection of horseshoe crab blood to endotoxin.

Limulus- genera of crab

Amebocytes- crab blood cell from which active component is derived.

Lysate- component is obtained by separating amebocytes from the plasma and lysing them.

Limulus polyphemus (horseshoe crab)

The basic procedure is the combination of 0.1 ml of test sample with 0.1 ml LAL Reagent.

After incubation for 1 hr at 370C, the mixture is analyzed for the presence of Gel clot.

The LAL test is positive, indicating the presence of endotoxin, if the gel clot maintains its integrity after slow inversion of the test tube containing the mixture.

This method has several advantages of Rabbit test like Greater sensitivity and reliabilityspecificity,lessvariation,widerapplication,less expensive and simplicity.

Gel/ clot reaction

Preparations for IV Fluids:

LVPs which are administered by IV route are commonly called as IV fluids.Purpose: Body fluids and electrolyte replenisherVolume supplied: 100 to 1000 mlAccording to their basic use, LVPS can be classified into:1-Basic nutrition.2-Restoration of electrolyte imbalance.3-Bodys fluid replacement.4-Blood and blood products.5-Drug carriers.6-Parenteral nutrition.7-Special (miscellaneous) use.

Examples of LVPs:Dextrose injection : Available in 2 , 5 , 10 , 25 & 50 % w/v solution.Used for: Fluids replenisher, Electrolyte replenisher Sodium chloride & Dextrose injection:Contains 0.11 to 0.9 % Sodium chloride and 2.5 to 5.0 % DextroseUsed for : Fluids replenisher, Electrolyte replenisher, Nutrient replenisher Sodium chloride injection IP:Contains 0.9 % conc., Also known as normal saline solution Used as : Isotonic vehicle , Fluids replenisher, Electrolyte replenisherSodium lactate injection IP:Contains 1.75 to 1.95 % w/v of sodium lactate Used as : Fluids replenisher, Electrolyte replenisher

Mannitol injection IP:Contains 5, 10 , 15, 20 % of mannitol Used as : Diagnostic aid, Renal function determination , as a diureticMannitol & Sodium chloride injection IP:Contains 5, 10 , 15, 20 % of mannitol & 0.45 % of Sodium chlorideUsed as :As a diureticRingers Injection, USP:Is a sterile solution of sodium chloride, potassium chloride and calcium chloride in water for injection (WFI).It is isotonic with physiological fluidsUsed as vehicle for other drugs or alone as electrolyte replenisher and plasma volume expander.Lactated Ringers Injection, USP:Lactated Ringer's solution is abbreviated as "LR" or "RL". It is also known as Ringer's lactate solution

Total Parenteral Nutrition (TPN)This is a complete form of nutrition, containing protein, sugar, fat and added vitamins and minerals as needed for each individual.Total Parenteral Nutrition (TPN) may be defined as provision of nutrition for metabolic requirements and growth through the parenteral route.

Components of TPN solutions:

(1) Protein as crystalline amino acids.(2) Fats as lipids.(3) Carbohydrate as glucose.(4) ElectrolytesSodium, potassium, chloride, calcium and magnesium.(5) Metals/Trace elementsZinc, copper, manganese, chromium, selenium.(6) Vitamins A, C, D, E, K, thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, choline and folic acid.

TPN might be necessary if:

A patient is severely undernourished, and needs to have surgery, radiotherapy or chemotherapy;

A patient suffers from chronic diarrhea and vomiting;

A baby's gut is too immature;

A patient's (their "gastrointestinal tract") is paralysed, for example after major surgery.

The solution is administered through superior venacava which is accessed by the subclavian vein near the heart.

The preferred method of delivering TPN is with a medical infusion pump.

A sterile bag of nutrient solution, between 500 ml and 4 L is provided. The pump infuses a small amount (0.1 to 10 mL/hr) continuously in order to keep the vein open.

Irrigation solutions:

They are topical solutions used to flush (clean) open wounds or body cavities.

Although they often labeled using the same terms for injections, they are never given parenterally.

Container size is usually larger than 1 liter.

Dialysis solutions:Dialysis is the process in which substances are separated from one another due to their differences in diffusibility (distribution) through the membrane.The fluids used in dialysis are known as dialysis fluids.

General uses Renal failure: to remove waste products and maintain electrolytes

Transplantation of kidney: Poisoning casesDialysis is a life saving procedure is done either by haemodialysis or intraperitoneal dialysis.

Haemodialysis is used to remove toxins from blood.

In haemodialysis, the blood from artery is passed through artificial dialysis membrane, bathed in dialysis fluid.

The dialysis membrane is permeable to urea, electrolytes & dextrose but not to plasma proteins & lipids. So excess of urea is passed out from blood through dialysis fluid.

After dialysis blood is returned back to the body circulation through vein.

A kidney unit may require more than 1200 litres of solution / week. So, haemodialysis fluid is prepared in concentrated form then it is diluted with deionised water or dist. water before use.Composition of Concentrated Haemodialysis Fluid BPC. Dilute 1 liter of conc. solution with 39 liters of water to make 40 litres.Storage: store in warm place as it is liable to convert into crystals on storage.

CompositionIntraperitonial dialysis is used to remove the toxic substances excreted by the kidney.

IV ADMIXTURES

Definition: When two or more sterile products are added to an IV fluid for their administration, the resulting combination is known as IV admixture.

In hospitals, prepared by nurses by combining or mixing drugs to the transfusion fluids.

The drugs may be admixed in one syringe or are incorporated into bottles of LV transfusion fluids.

Must be prepared under ASEPTIC conditions.