Main Speaker: Dr. Pankaj Aggarwal, Pfizer, Inc. Groton, CT 06340

Title: Computational approaches for Chromatographic Method Development in Pharmaceutical Industry

Abstract: Systematic approaches to chromatographic method development with emphasis on knowledge gathering and method control provide robust separation methods with fewer analysis failures. The chromatographic modeling tools capable of multi-factor optimization can reduce the amount of experimental work needed and simultaneously deliver globally optimum method conditions. This work presents the development of a robust UPLC method using multi-factorial optimization approaches for analyzing the purity and potency of a pharmaceutical dosage form. Commercially available chromatography modeling software (such as DryLab, Design Expert, ACD Labs) was employed to determine the final method conditions and evaluate the method robustness.

Bio: Dr. Aggarwal is currently a Senior Scientist at Pfizer in Groton, CT.  He received his Ph.D. degree in Analytical Chemistry from the Milton L. Lee lab at Brigham Young University. His thesis work focused on developing high efficiency monolithic stationary phases for capillary HPLC. During this period, he developed a strong fundamental and practical understanding of column and methods development for HPLC. Pankaj joined Pfizer in December 2014 in the QbD method development group within ARD. In this role, Pankaj has been developing analytical methods for HPLC, UPLC, and GC by applying QbD principles and computational approaches to support drug development.  He currently serves as a member on the Board of Directors  with the Connecticut Separations Science Council (CSSC).

 

Graduate Student Speaker: Ms. Di Jiang - Rusling Lab, University of Connecticut (Poster award recipient - Csaba Horvath Memorial Award Symposium 2016)

Title: Sequence specific DNA oxidative damage on p53 gene fragments for determination of tissue specific cancers using LC-MS/MS

Abstract: DNA oxidation, specially related to guanine oxidation, is mutagenic which is important in carcinogenesis. The oxidation product, 8-oxo-7,8-dihydroguanosine, may lead a G to T transversion mutation. P53 tumor suppressor gene is known to be mutated in more than 50% of human cancers. From the p53 database, mutation sites on p53 gene are correlated with specific types of cancers. If the most frequently mutation codons on p53 gene in vitro coincide with the known p53 hot spots in vivo, organ specific cancers would be predicted. Here, we used catechol in presence of redox cycling system (NADPH and Cu2+), which generates reactive oxygen species (ROS), to oxidize 32 base pair (bp) oligonucleotide from part of exon 7 on p53 gene with 7 possible hot spots. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to sequence the oligonucleotides from oxidized 32 bp fragments which were fragmented by a restriction endonuclease and quantify the positions of oxidized DNA bases. Analysis of oxidized fragments, codons 243, 244, 245, 246 and 249 are most frequently mutated codon sites. Codon 245 is mutated in colorectal cancer, while codons 244, 249 are mutated in non-small cell and small cell lung cancers, which supporting a possible role of DNA oxidation damage in the lung cancer.

Bio: Ms. Di Jiang is a third year graduate student in Dr. Rusling’s group at the  University of Connecticut. She received her Bachelor’s Degree in Chemistry from Jilin University in China in 2011. She received her Master’s Degree from Jilin University, in 2014. She then came to the United States for her Ph.D. studies. Her current work is on sequence specific DNA oxidative damage on the p53 tumor suppressor gene fragments by using LC-MS/MS.  She is a  poster award recipient from the Csaba Horvath Memorial Award Symposium held at the Yale University Conference Center in October of 2016.