Alcala Pharmaceutical, Inc.

Matthew D Rifat

David J Smith

Christian Tagwerker

Ranulu Samanthi De Zoysa

Eric J Brunson

Irfan Baig

A method for extracting pharmacogenetic DNA from dried blood that is retained in a solid resin collection device includes one or more of the steps of combining the dried blood and at least a portion of the solid resin collection device in a first vessel, sonicating the first vessel containing the dried blood and at least a portion of the solid resin collection device, moving at least some of the contents of the first vessel including the portion of the solid resin collection device to a solid resin spin column, adding an elution buffer to the solid resin spin column, centrifuging the solid resin spin column so that at least some of the contents of the solid resin spin column are transferred to a second vessel, and processing the contents of the second vessel by conducting one of capillary electrophoresis, Next-Generation Sequencing, DNA sequencing or genotyping, and mass spectrometry-based sequencing on the contents of the second vessel.

Method for extracting pharmacogenetic DNA from biological fluid retained in a solid resin collection device

A method for preparation of a dried blood sample for multiplexing of analytes includes the steps of mixing an Internal Standard solution with a first diluent in a vessel, the Internal Standard solution including a plurality of different Internal Standards, adding the dried blood sample to the vessel, sonicating the vessel containing the Internal Standard solution, the first diluent and the dried blood sample, and removing the dried blood sample from the vessel. The Internal Standard solution can include a plurality of Internal Standards. The Internal Standard solution can include at least 15 Internal Standards. The dried blood sample can be generated using less than 50 &#x3bc;L of blood. The dried blood sample can be generated using less than 10 &#x3bc;L of blood. The first diluent can include methanol or a mixture of water and methanol.

Method for preparation of dried blood sample for multiplexing of analytes

A method for qualitative and quantitative multiplexing of drug analytes from dried blood samples includes the steps of mixing an Internal Standard solution with a first diluent in a vessel, adding the dried blood sample to the vessel, sonicating the vessel containing the Internal Standard solution, the first diluent and the dried blood sample, removing the dried blood sample from the vessel so that a final sample can be attained, and analyzing at least a portion of the final sample using one of LC-MS and LC-MS/MS to simultaneously determine the presence or absence of a plurality of different analytes in the dried blood sample. The Internal Standard solution can include at least 8, 15, 25 or 50 Internal Standards. The dried blood sample is generated using less than 50 &#x3bc;L, 20 &#x3bc;L or 10 &#x3bc;L of blood. The step of analyzing includes simultaneously determining the presence or absence of at least 15, 60, 90 or 130 different analytes. A ratio of the number of analytes for which the presence or absence is being determined to the number of Internal Standards in the Internal Standard solution is at least approximately 2:1. A ratio of the number of analytes for which the presence or absence is being determined to the volume (in &#x3bc;L) of the final sample being analyzed is at least approximately 2:1. A ratio of the number of analytes for which the presence or absence is being determined to the volume (in &#x3bc;L) of blood from which the dried blood sample was obtained is at least approximately 4:5.

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San Diego, CA, United States of America

Matthew D Rifat