TABLE 1

Detailed Protocol for BET

Test serial no.	Assay tube (in duplicate)	LAL reagent water (μL)	Control standard endotoxin 4λ (μL)	Test sample^* (μL)	LAL reagent (μL)
1	Negative water control	100	—	—	100
2	Positive water control 2λ	50	50	—	100
3	NPC^†	50	—	50	100
4	PPC 2λ^†	—	50	50	100
5	PPC 2λ with calcium chloride^†	—	50	50^‡	100

↵* Repeated at MVD and MVD/2—that is, 1:66 and 1:33 dilutions.
↵† These tubes were set up for all ^99mTc radiopharmaceuticals (^99mTc-DTPA, ^99mTc-DMSA-III, ^99mTc-DMSA-V, ^99mTc-ECD, ^99mTc-mebrofenin, ^99mTc-methoxyisobutylisonitrile, and ^99mTc-MDP) and their cold kits.
↵‡ Calcium chloride at final concentration of 300 μM was incorporated in assay tube of test sample at final dilution.
Tests 3 and 4 were also performed initially for 60 mM stock solution of calcium chloride and saline (0.9% sodium chloride) to ensure absence of endotoxin in stock solutions.