8. Results

Use Microsoft word for further calculations and results. Make standard curve and endotoxin concentration (Figure 3).

8.4 Advantages of Gel Clot method

DOI: http://dx.doi.org/10.5772/intechopen.81331

This is very easy to perform. This is not time consuming. Accuracy is 100 percent.

over a range of concentrations.

at 2–8°C. Storage life is 15 days.

8.5.4 For water for injection

We have to make dilution.

Results shown sample pass or not.

means gel has not formed. This means negative.

8.5.1 Material

(LRW 50 ml).

8.5.3 Procedure

control [1].

(Table 4):

Table 4.

101

8.5.5 For product

8.5.2 Reconstitution

8.5 Standard operating procedure

Gel Clot LAL provides a simple positive/negative result and is most often men-

What Is Limulus Amebocyte Lysate (LAL) and Its Applicability in Endotoxin Quantification…

The LAL Gel-Clot assay, gives a more quantitative measurement of endotoxin

Gel Clot lysate for 20 test, Gel Clot standard 0.5 EU/Vial, LAL reagent water

Lysate: add 2 mL LRW and mix it slowly. Do not shake and avoid foaming.

Take three test tubes and mark them as test, positive control and negative

Add your sample in test tube marked as sample. Add standard in test tube marked as Positive control. Add LRW in test tube marked as negative control. Incubate the test tubes at 37 + 2°C for 60 min. After an incubation, check for the gel by inverting the test tube. If the material remains firm in the bottom of the test tube, it means gel has formed. This positive if the material gets the flow down, it

Take similarly three test tubes as above and add water for injection (WFI) in test

tube marked as sample. And proceed as above. The results should be as follows

Sample Positive control Negative control Result ve (gel not formed) +ve ve Pass Sample Positive control Negative control Result +ve (gel formed) +ve ve Fail

Standard: Add 2 mL of LRW in the vial and mix it well for 15 min. Store the vial

Transfer 0.1 ml in 20 test tubes. Store it at –degree (in freezer).

tioned in pharmacopeial monographs as the official referee test.

R2 = coefficient of determination R = correlation coefficient R ≥ 0.98 R2 = 0.99 R = √R2 = 0.99 Equation of straight line y = mx + c m = slop x = endotoxin concentration c = y intercept y = mean absorbance Equation of straight line Y = 1.019X 0.026
