Solution | Endotoxin Concentration/Solution to which Endotoxin is Added | Diluent | Dilution Factor |
Initial Endotoxin Concentration |
Number of Replicates |
Aa | none/sample solution | | | | 4 |
Bb | 2/sample solution | sample solution | 1 2 4 8 |
2 1 0.5 0.25 |
4 4 4 4 |
Cc | 2/water for BET | LAL Reagent Water | 1 2 4 8 |
2 1 0.5 0.25 |
2 2 2 2 |
Dd | none/LAL Reagent Water | | | | 2 |
a
Solution A: a sample solution of the preparation under test that is free of detectable endotoxins.
b
Solution B: test for interference.
c
Solution C: control for labeled LAL Reagent sensitivity.
d
Solution D: negative control of LAL Reagent Water.
|
Solution* | Endotoxin Concentration/Solution to which Endotoxin is Added | Number of Replicates |
A | none/diluted sample solution | 2 |
B | 2/diluted sample solution | 2 |
C | 2/LAL Reagent Water | 2 |
D | none/LAL Reagent Water | 2 |
*
Prepare Solution A and positive product control Solution B using a dilution not greater than the MVD and treatments as directed in the Interfering Factors Test for the Gel-Clot Techniques under Preparatory Testing for the Gel-Clot Techniques. Positive control Solutions B and C contain the standard endotoxin preparation at a concentration corresponding to twice the labeled LAL Reagent sensitivity. The negative control Solution D is LAL Reagent Water.
|
Solution | Endotoxin Concentration/ Solution to which Endotoxin is Added |
Diluent | Dilution Factor | Initial Endotoxin Concentration |
Number of Replicates |
Aa | none/sample solution | LAL Reagent Water | 1 2 4 8 |
|
2 2 2 2 |
Bb | 2/sample solution | | 1 | 2 | 2 |
Cc | 2/LAL Reagent Water | LAL Reagent Water | 1 2 4 8 |
2 1 0.5 0.25 |
2 2 2 2 |
Dd | none/LAL Reagent Water | | | | 2 |
a
Solution A: a sample solution under test at the dilution, not to exceed the MVD, with which the Interfering Factors Test for the Gel-Clot Techniques was completed. Subsequent dilution of the sample solution must not exceed the MVD. Use LAL Reagent Water to make dilution series of four tubes containing the sample solution under test at concentrations of 1, ½, ¼, and 1/8 relative to the dilution with which the Interfering Factors Test for the Gel-Clot Techniques was completed. Other dilutions may be used as appropriate.
b
Solution B: Solution A containing standard endotoxin at a concentration of 2 (positive product control).
c
Solution C: two series of 4 tubes of LAL Reagent Water containing the standard endotoxin at a concentration of 2, , 0.5, and 0.25, respectively.
d
Solution D: LAL Reagent Water (negative control).
|
Solution | Endotoxin Concentration | Solution to which Endotoxin is Added | Number of Replicates |
Aa | none | sample solution | not less than 2 |
Bb | middle concentration of the standard curve | sample solution | not less than 2 |
Cc | at least 3 concentrations (lowest concentration is designated ) |
LAL Reagent Water | each not less than 2 |
Dd | none | LAL Reagent Water | not less than 2 |
a
Solution A: the sample solution may be diluted not to exceed MVD.
b
Solution B: the preparation under test at the same dilution as Solution A, containing added endotoxin at a concentration equal to or near the middle of the standard curve.
c
Solution C: the standard endotoxin at the concentrations used in the validation of the method described in Verification of Criteria for the Standard Curve under Preparatory Testing for the Photometric Techniques (positive control series).
d
Solution D: LAL Reagent Water (negative control).
|