I have been struggling with B12 (cyanocobalamin) Micro Assay which uses Lactobacillus Leichmannii.

I am using USP 171 as the reference method. I'm not quite understood of the procedure.

I'm not getting the results. As we are suspecting the growth of Lactobacillus is not strong as it gives high transmittance reading when I cross check with UV-Vis.

How do I stab transfer of pure culture of Lactobacillus Leichmannii, and make not fewer than 10 successive transfers of the culture in a 2- week period?

Can someone help me with this L. Leichmannii cultivation and the assay. As the referance as follows:

USP REFERANCE STANDARDS Vitamin B12 Activity Assay.

Thank You in advance.

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