THINH A VETERINARY MEDICINE PRODUCTION AND TRADING

JOINT STOCK COMPANY (ASIFAC)

QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

Written by Quality Control Manager Approved by Site Manager

Date: January 2022 Date: January 2022

Ta Thi Hao Nguyen Le Thanh Tuan

Distribution
 QC □ Stock
 QA □ Maintenance
□ R&D □ Marketing
□ IPC □ Production
□ Logistics

History of modifications

Amendment/addition
Edition N° Reason for amendment/addition
of page number
1 January 2020 Revision of the procedure

2 January 2022 Revision of the procedure

Page 1 of 6
 THINH A VETERINARY MEDICINE PRODUCTION AND TRADING
JOINT STOCK COMPANY (ASIFAC)
QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

1. Objective
To prove that the analytical method of Lincomycin in ASI-LINCO 4.4% is suitable for its
intended use.

2. Scope
Analytical method of Lincomycin in ASI-LINCO 4.4%.

3. Responsibility:
Methodology section head is responsible for implementation of the protocol.
QA manager is responsible for reviewing and approval of the protocol and the final report.

4. Composition:
Active Ingredients Quantity per 1000g
1 Lincomycin 44 g

5. Documentation required:
Analytical monograph for ASI-LINCO 4.4%.
Equipment operation procedure.

6. Analytical performance characteristics and acceptance criteria:

STT Parameters Acceptance criteria
1 Linearity and range r2 ≥ 0,995
2 Accuracy and recovery 97% – 103%
3 Precision and repeatability RSD ≤ 2,0%
4 Specificity/Selectivity Pass the specificity/Selectivity test
Pass Accuracy and recovery test → Pass
5 Roughness
Roughness test

Page 2 of 6
 THINH A VETERINARY MEDICINE PRODUCTION AND TRADING
JOINT STOCK COMPANY (ASIFAC)
QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

7. Revalidation:
It is necessary in the followwing circumstances:
- Changes in the composition of finished product.
- Changes in the analytical procedure.

8. Procedure:
8.1 Linearity and range:
- Linearity shows the correlation between sample concentration and absorbance of the device.
System precidion is conducted using 5 replicates of the standard solution.
Operational conditions
o Solution A: Dissolve 13.5ml of H3PO4 in 1000 mL of distilled wate, adjust pH =
6 with Triethylamine.
o Mobile phase : Acetonitrile : Solution A (15 : 85).
o Column : C18 5µm (4.6mm x 150mm)
o Flow rate : 1.0 ml/min.
o Volume injected : 20µl
o Wavelength : 210 nm
o Machine stabilization time: 30 minutes
- Preparation of solutions: Dissolve 0.05 g of reference Lincomycin in the mobile phase and
complete to 50.0 ml.
- Prepare a straight line: The standard solution were transferred in turn to a 10 ml volumetric
flask as shown in the following table and diluted to 10 ml with the same solvent. Filter
membrane 0.45µm.
The standard solutions (ml) Volume (ml)

30% 3 10

Page 3 of 6
 THINH A VETERINARY MEDICINE PRODUCTION AND TRADING
JOINT STOCK COMPANY (ASIFAC)
QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

40% 4 10
50% 5 10

60% 6 10

70% 7 10

- From the obtained statistics, determine the correlation coefficient r2, build a regression
equation and plot a linear line. Coefficient r2 ≥ 0,995.
- For each concentration, conduct 2 times → the average result.
8.2 Precision and repeatability:
- Determination of content in 6 independently phased test samples of the finished lot.
- Standard: Dissolve 0.05 g of reference Lincomycin in the mobile phase and complete to
100.0 ml. Filter membrane 0.45µm.
- Sample: Dissolve 1 g of product in the mobile phase and complete to 100.0 ml. Filter
membrane 0.45µm..
Calculate the quantity in g/kg, of Lincomycin by the formula:

Where:
mstd: Mass of standard substance (mg)
msmp: Mass of sample (mg)
Sstd: Area of chromatography peak of standard.
Ssmp:Area of chromatography peak of sample.
T: Titer of standard substance.
k: Dilution factor of sample and standard
- Request: Precision and repeatability RSD ≤ 2,0%

Page 4 of 6
 THINH A VETERINARY MEDICINE PRODUCTION AND TRADING
JOINT STOCK COMPANY (ASIFAC)
QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

8.3 Accuracy and recovery:

- Accuracy and recovery is performed on 5 independently phased test samples of a batch of
finished product.
- Sample: Dissolve 1 g of product in the mobile phase and complete to 100.0 ml and add an
amount of the standard not less than 10% of the analyte content in the sample. Filter membrane
0.45µm..
- The recovery coefficient is calculated according to the formula:

Sa T
Y (mg) =  mchuan  k
Schuan 100

X
Y − (mmau  )  100
1000
A% =
mi
- Trong đó:
➢ Y – Total content of standard addition sample (mg).
➢ X – Actual content (g/kg).
➢ Sa – Sample area and added standard.
➢ Schuan – Standard area.
➢ T – Standard content (%).
➢ k – Dilution factor.
➢ mmau – Sample mass (mg).
➢ mi – Standard mass added (mg).
➢ mchuan – Standard mass (mg).

8.4 Specificity/Selectivity
Page 5 of 6
 THINH A VETERINARY MEDICINE PRODUCTION AND TRADING
JOINT STOCK COMPANY (ASIFAC)
QUALITY CONTROL DEPARTMENT
VALIDATION OF ANALYTICAL PROCEDURES
OF ASI-LINCO 4.4% POWDER
Version n 3 Number of documents: QC – AVP023

- The specificity/Selectivity of the method is evaluated to ensure that there is no influence

on the quantitative results from impurities and excipients in the formulation.
- Determine the absorbance of a self-made sample containing the same excipients as the test
solution but without the active ingredientLincomycin. From there conclude the
Specificity/Selectivity of the analytical method.
8.5 Roughness:
- Pass Accuracy and recovery test → Pass Roughness test.

Page 6 of 6