Analysis method of Alpha Beta Arteether Injection

1.0 OBJECTIVE
1.1 To lay down a procedure for Analysis of Alpha Beta Arteether Injection

2.0 SCOPE
2.1 This procedure is applicable to the analysis of Analysis of Alpha Beta Arteether Injection in QC lab

3.0 RESPONSIBILITY
3.1 Q.C- Chemist

4.0 ACCOUNTABILITY
4.1 Manager-Quality Assurance

5.0 PROCEDURE
5.1 Identification:
5.2 Description: Pour 50ml finish sample in Beaker and observed Visually.
5.3 pH: Taken 50ml sample in beaker rinse the pH electrode first with purified water followed by sample dip the electrode in sample and observe the pH.
5.4 Volume variation: Measured the volume by syringe and determine the volume variation.
5.5 Sterility:
MEMBRANE FILTRATION METHOD
5.5.1 Wipe the sample article individually with 70% IPA solution and keep in a clean S.S trays marked with Product Name, Batch No. and Lot No, and then transfer the samples to the sterility room through clean pass box for performing sterility.
5.5.2 Prepare the media tubes (FTM and SCDM) as per the SOP for preparation of culture media
Dispense 100 ml quantity for membrane filtration and for Direct Inoculation Method. Sterilize both
the media at 1210C and 15 psi pressure for 20 minutes as per SOP for Media Sterilization by Autoclaving.
5.5.3 After autoclave Label the tubes with Name of Media, Media Batch No. and pre-incubate the media tubes at appropriate temperature i.e. SCDM tubes at 20 to 250C whereas FTM tubes at 30 to 350C for 24 – 48 hrs. before subjecting them for sterility operations.
5.5.4 Autoclave Dress, Scissors, forceps and Filtration unit in a S.S Container at 1210C temperature and 15psi pressure for 30 minutes.
5.5.5 After Sterilization cool the contents and aseptically transfer in a S.S. container to cleaned Pass box.
5.5.6 Transfer the pre incubated sterile media tubes, SCDA plates, sterile swabs, sterilized Filtration unit, and 0.1% w/v peptone water to the sterility test room through pass box.
5.5.7 Enter in sterility room as per the Entry / Exit procedure for Sterility Room.
5.5.8 Start the LAF as per SOP.
5.5.9 Wipe out all samples to be tested for sterility with 70% filtered IPA solution.
5.5.10 Before starting sterility test, expose the SCDA plates as specified locations throughout the testing.
5.5.11 Connect the Glass filtration cup holder with the Filtration flask properly with pipe under laminar airflow unit.
5.5.12 Check the Manometer reading of working LAF and check the temperature as well as humidity of the sterility room.
5.5.13 Switch ON the vacuum pump with the help of switch, present on the wall. Now place 0.45 sterile membrane filters between filtration cup and receptacle with the help of sterilized forceps and clamped it properly.
5.5.14 Wet the membrane filter by adding approx. 15 ml of sterilized Fluid A (0.1% peptone water) to filter holder, and filter the fluid by employing vacuum.
5.5.15 Cut the tip of bottle/vial or ampoule with sterile SS blade in front of the gas burner and immediately transfer the contents of sample to membrane filter. For dry powder or lyophilized container, add the sterile water 0.1% peptone dissolve and then collect it in sterile flask and immediately transfer the contents of sample to membrane filter.
5.5.16 Immediately filter the solution with the aid of vacuum and wash the membrane three times with 100 ml of sterilized fluid A (0.1% peptone water).
5.5.17 After complete filtration, stop the vacuum pump.
5.5.18 Lift the membrane carefully with the help of sterile forceps, aseptically cut the membrane filter into two halves with sterile SS scissor and transfer one half to FTM and one half to SCDM tubes by unplugging in front of gas burner only.
5.5.19 Label both the tubes with Product name, B. No, Report No.., date of testing, Completion date & Tested by.
5.5.20 Simultaneously prepare a negative control by filtering 100 ml of 0.1% peptone water
5.5.21 Instead of product sample, cut the membrane into two halves with sterile SS scissor and transfer one half to FTM and one half to SCDM and label both the tubes as Negative control.
5.5.22 For media negative control, keep one – one tube of each autoclaved lot of FTM & SCDM un-inoculated.
5.5.23 After completion of work transfer all inoculated media through hatch box and then transfers all the equipment and exposed plates to microbiology analysis section.
5.5.24 Incubate the FTM tubes at 300C – 350C and SCDM tubes at 200C – 250C. Incubation period for terminally sterilized products are 7 days and 14 days for aseptically filled products as per IP. If the Product is as per USP, BP, incubation period is 14 days for both terminally sterilized as well as for aseptically filled products.
5.5.25 Start the LAF of Biosafety cabinet as per SOP and prepare three positive Control tubes by inoculating aseptically not more than 100 cfu in FTM tubes with S. aureus, P. aeruginosa and Clostridium sporogenes. Similarly prepare three SCDM positive control by inoculating not more than 100 cfu separately with C. albicans, A. niger, and Bacillus subtilis. Incubate FTM positive control tubes at 30 – 35ºC & SCDM positive control tubes at 20 – 25ºC. For bacterial positive controls, incubation period is not more than 3 days & for fungal positive control, incubation period is not more than 5 days.
5.5.26 Bacterial endotoxin test
PROCEDURE FOR TESTING:

5.6.3 Dilute the sample MVD/2 times with LAL water when performing the test at MVD. If performing the test at MVD/2, dilute the sample MVD/4time with LAL water.

5.7 Preparation of E-Coli Control Standard Endotoxin (CSE):
5.7.1 Reconstitute the CSE with appropriate volume (as stated on certificate) of LAL reagent water vortex continuous 30 minutes at the interval of 10minutes (When new bottles reconstitute) & vortex vigorously for five minutes before further dilution
5.8 Preparation of CSE dilution series:
5.8.1 Confirm the labeled sensitivity using at least one vial of each batch/lot of lysate. Prepare a series
of two-fold dilution of the CSE to give concentrations of 2, , 0.5 and 0.25, where  is the labeled sensitivity of the lysate in EU per ml.
5.8.2 Perform the test as given as under Procedure on these four standard concentrations in duplicate or greater in include negative control consisting of water BET.
5.8.3 Calculate the average of the logarithms of the lowest concentration of endotoxin in each series of dilutions for which a positive result is found. The antilogarithm of this average gives the estimated lysate sensitivity, which must be greater than or equal to 0.5 and less than or equal to 2.0.
confirm the labeled sensitivity of each new batch of lysate prior to use in the test. Test CSE results are only valid when the positive water and specimen controls are positive at the 2  &  endotoxin concentration.

5.9 Preparation of LAL Reagents:
5.9.1 Test LAL reagent for its sensitivity at the time of receiving. For reconstitution, Collect LAL powder into the bottom of the vial by tapping on a firm surface, unseal and release the vacuum by slowly lifting the stopper, avoiding touch contamination. The small amount of LAL on the stopper is insignificant.
5.9.2 Rehydrate the reagent with appropriate volume of (as stated on label and certificate) LAL reagent water by pipetting directly into the vial immediately before use. Cover the vial with an endotoxin free surface or
the inner side of perafilm when not in immediate use. Gently swirl until LAL dissolves into a colorless solution.
5.10 Test procedure for gel clot:
5.10.1 Each assay should include dilutions of a sample or product, a negative water control, a positive product control, and either a 2 water control or a series of two-fold dilutions from an endotoxin standard which bracket the labeled LAL sensitivity.

5.11 Interpretation of results:
5.11.1 Each tube in the gel-clot method is interpreted as either positive or negative. A positive test is defined as formation of a firm gel capable of maintaining its integrity when the test tube is inverted 180°C. A negative test is characterized by the absence of gel or by the formation of a viscous mass, which does not hold when the tube is inverted. Calculate sample endotoxin concentration by the following formula.
Sample Endotoxin Conc. :- Dilution Factor x Sensitivity of 
Conc. of Sample (Potency)

Sr. No.	Abbreviation used.	Full form of abbreviation used.
1.0	STP	Standard Testing Procedure.
2.0	QA	Quality assurance.
3.0	STD	Standard.
4.0	SPL	Sample
5.0	I.P.	Indian Pharmacopeia
6.0	NM	Nano meter

Sr. No.	Reference Title
01	In House

Analysis method of Alpha Beta Arteether Injection

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