QUALITY CONTROL TESTS OF CYTOTOXIC DRUGS (CYTARABINE)

FRESENIUS KABI ONCOLOGY (DABUR PHARMA LTD.) SOLAN,HIMACHAL PRADESH

PRESENTED BY: PREETIKA RATHOUR ROLL NO. 301001018

QUALITY CONTROL OF CYTOTOXIC DRUGS

Quality can be defined as status of drug that is determined by identity, purity, content and other chemical, physical or biological properties of the drug. Quality control refers to processes involved in maintaining the quality and validity of manufactured product.

It is the process or system for monitoring the quality of laboratory testing, and the accuracy and precision of results. Data is routinely collected and analyzed from every test run or procedure.It allows for immediate corrective action. It is responsible for the acceptance and rejection of the incoming raw materials,packaging components,in process tests and finally for approved or rejection of finished products.

Microbiology QC: To check sterility,Ability to

support growth,Selective or inhibitory characteristics of the medium and to test QC organisms with each new batch or lot number to check for growth of fastidious organisms on media of choice and incubate at time and temperature recommended

CYTOTOXIC DRUGS:
Cytotoxic drugs (CDs) are used as anti- cancer drugs.They kill cancer cells or stop them from multiplying in different ways. However, they all tend to work by interfering with some aspect of cell division and multiplication. Cytotoxic drugs are effective where cells are rapidly dividing.However, some normal cells in the body divide and multiply quite rapidly like hair cells, bone marrow cells, and cells lining the mouth and gut,these may be affected by cytotoxic drugs.

Ara-C (Arabinofuranosyl Cytidine) injection

Cytarabine, or Cytosine arabinoside, is a chemotherapy agent used mainly in the treatment of cancers of white blood cells such as acute myeloid leukemia or non Hodgkin lymphoma. The empirical formula is C9H13N3O5 and has a molecular weight of 243.217 g/mol. Cytarabine is an odourless,white to off-white, crystalline powder is freely soluble in water and slightly soluble in alcohol and in chloroform.

PHARMACODYNAMICS
Cytarabin destroys cancer cells by interfering with DNA synthesis. Cytosine normally combines with deoxyribose, to form deoxycytidine. Cytosine arabinoside is similar enough to cytosine deoxyribose (deoxycytidine) to be incorporated into human DNA, but different enough that it kills the cell. It inhibits both DNA and RNA polymerases and nucleotide reductase enymes needed for DNA sythesis.

DIFFERENT INSTRUMENTS USED TO CHECK QUALITY

1.

pH METER: It is used to determine the acidity or alkalinity of the solution.

According to the defined standards or the technical information of Cytarabine,pH should be from 7 to 9.
The test performed for pH of cytarabine with concentration of 20mg/ml,its pH range was found out to be 7.2-8.0.

2. AUTO KARL-FISCHER TITRIMETER:

Karl fischer apparatus is used for determination of the moisture of the drug or determination of water. The anodic compartment contains the anolyte solution which includes sulfur dioxide (SO2), iodide (I-) and imidazole needed in the chemical reaction. Methanol or ethanol (ROH) is usually used as a solvent. ROH+SO2+RN (RNH).SO3R

RNH.SO3R+2RN+I2+H2O (RNH).SO4R+2(RNH)I 2II2+2e-

Water (%W/V) =V*F*100/W Where, V=volume of the reagent consumed (in ml) F= water equivalence factor of the reagent (mg/ml) W= weight of sample taken in mg Determination of water using KF (Karl Fischer) coulometric titration for determining free water and water of hydration in most solid or liquid organic and inorganic compounds was found to be .05% -1%. The KF of samples found to be 0.50% ,within the specified limits.

3. FOURIER TRANSFORM INFRA RED SECTROPHOTOMETER

FTIR is most useful for identifying chemicals and is most powerful tool for identifying types of chemical bonds (functional groups).

The wavelength of light absorbed is characteristic of the chemical bond. By interpreting the infrared absorption spectrum, the chemical bonds in a molecule can be determined.

Wavenumber (cm-1)

4. UV SPECTROPHOTOMETER
UV Spectrophotometers can be divided into 2 ranges: -Far UV or under vacuum: 185-200nm -Near UV without vacuum: 200-400nm
When radiation is passed through a layer of the solution containing an absorbing substance, part of the radiation is absorbed, the solution is less than the intensity of the radiation. The magnitude of the absorption is expressed as: A=log10 (I0/I)

For the selection of analytical wavelength, 15g/mL solution was prepared by subsequent dilutions and was scanned in the spectrum mode from 200nm400nm. The peak height of Cytarabine was at its maximum at 280 nm.

5.High Performance Liquid Chromatography

Separation of components from one another by the column packing that involves various chemical and/or physical interactions between their molecules and the packing particle.

These separated components are detected at the exit of the column by a detector that measures the amount. An output from this detector is called a liquid chromatogram.
Here separation is more effective due to greater surface area achieved due to very small particle size of stationary phase.

HPLC GRAPH OF STANDARD

The Retention time was found to be 11.43 minutes.

HPLC GRAPH OF UNKNOWN

The Retention time was found to be 11.45 minutes.

The representative chromatogram of standard solution alone and that containing 15ml of Cytarabine showed almost same retention time.
Thus, absence of any overlapping or extraneous peaks in both chromatograms indicates its specificity. Area Purity Standard 9180640 100% Sample 8670644 94.44%

Thus, as the % purity was almost close to the standard, we could say that our sample of Cytarabine was pure.

6.Test for Visible and Sub visible particles:

TEST Visible particles RESULT Solution is free from visible particles SPECIFICA TION Practically free from visible particles.

Sub-visible particles >=10m 17partilces per vial Not more than 6000 per vial.

>+25m

1 particles per vial

not more than 600 per vial.

Various suspended particles are observed under the beam of light and if too many subvisible particles between size 10 m to 25 m then batch is rejected.

6.MICROBIOLOGICAL QC
When performed a microbiological assay for any strains present in the vial, no turbidity was observed .
Samples from vials are serial diluted and then plated on different media and no growth was observed.

CONCLUSION
Thus the above tests performed helps in survey of the market value of the drug and pharmaceutical company and stability,which explains its national as well as international market, in terms of its therapeutic effects