Effect of the refrigerator storage time on the potency of botox for human extensor digitorum brevis muscle paralysis

Abstract

Background and purpose: It is recommended that Botox be used within 5 hours of reconstitution, which results in substantial quantities being discarded. This is not only uneconomic, but also inconvenient for treating patients. The aim of this study was to determine the potencies of Botox used within 2 hours of reconstitution with unpreserved saline, the same Botox refrigerated (at +4℃) 72 hours after reconstitution, and during the next 4 consecutive weeks (weeks 1, 2, 3, and 4). This comparison was used to determine the length of refrigeration time during which reconstituted Botox will maintain the same efficacy as freshly reconstituted toxin.

Methods: Individual paralysis rates in the extensor digitorum brevis (EDB) compound muscle action potential (CMAP) amplitude and area were measured 1 week after injecting fresh reconstituted 2.5 MU of Botox on one side of the foot, and when the same quantity of Botox that had been refrigerated for a designated time (i.e., 72 h, or 1, 2, 3, or 4 weeks) into the other side of the foot. The EDB CMAP amplitude and area at 12 and 16 weeks postinjection were also measured to compare the efficacy durations in all five comparative groups.

Results: Ninety-four volunteers were divided into five groups according to the refrigerator storage time of the second Botox injection. The paralysis of the EDBs was significant for each injection of Botox, both fresh and refrigerated, with no statistically significant differences between them, regardless of the refrigeration time. There was a tendency toward increased CMAP amplitude and area at 12 or 16 weeks postinjection (p<0.0001). The duration of effective muscle paralysis did not differ significantly throughout the 16-week follow-up period between all five groups.

Conclusions: The potency of reconstituted Botox is not degraded by subsequent refrigeration for 4 weeks. However, there are definite concerns regarding its sterility, and hence its safety, since multiple withdrawals from the same vial over long periods can introduce bacterial contamination.

Keywords: botox; potency; refrigeration.

Fig. 1

Study design. EDB CAMP: extensor digitorum brevis compound muscle action potential, hrs: hours, Lt.: left, Rt.: right, wk: week.

Fig. 2

Extensor digitorum brevis (EDB) muscle compound muscle action potential amplitudes and areas of representative subject who was injected 2.5 MU/0.1 mL fresh Botox and 1 week refrigerated Botox on each side of the EDB muscles. There are meaningful EDB paralysis with both fresh and refrigerated toxins at 1, 12, and 16 weeks post injection.