Presidential Green Chemistry Challenge: 2013 Greener Synthetic Pathways Award

Life Technologies Corporation (technology acquired by Thermo Fisher Scientific)

Safe, Sustainable Chemistries for the Manufacturing of PCR Reagents

Innovation and Benefits: Polymerase Chain Reaction (PCR) is the process used to perform genetic testing. Manufacturing the key chemicals required for PCR tests is quite wasteful, often producing thousands of times more waste than product. Life Technologies has developed a three-step, one-pot synthesis which is much more efficient. The new process prevents about 1.5 million pounds of hazardous waste a year.

Summary of Technology: Polymerase chain reaction (PCR), the process of amplifying genetic material, is used in basic research, genetic engineering, forensics, infectious disease identification, food safety, and, most recently, personalized medicine. The expanding role of PCR in science and medicine highlights the need to create safe and sustainable chemistries for the manufacture of reagents for these applications. Deoxyribonucleotide triphosphates (dNTPs) are the individual building blocks for the DNA that is made during PCR. Conventional syntheses of dNTPs are inefficient, involve multiple steps that require isolation and purification of intermediates, and use excessive volumes of toxic or hazardous solvents and reagents.

Researchers at Life Technologies have devised synthetic routes for the manufacture of dNTPs that are only three steps in a single pot, eliminating the need to transfer reaction material. These synthetic routes also eliminate the need for a variety of hazardous reagents and solvents, including zinc chloride, triphenyl phosphine, aldrithiol, dimethyl formamide (DMF), and dichloromethane. By using these new dNTP routes, worker exposure to hazardous materials is minimized and the process E-factor (the ratio of amount of waste to amount of product) is improved by about a factor of 10.

In 2011, Life Technologies implemented these greener synthetic routes for the full-scale production of dNTPs and their analogues at their Austin, Texas manufacturing site. Organic solvent consumption has been reduced by up to 95 percent and other hazardous waste up to 65 percent compared to conventional protocols. By improving the yields and specificity of reaction, process E-factor (the ratio of the mass of waste to the mass of product) has been reduced from approximately 3200 to 400, almost a full order of magnitude, leading to a savings of 1.5 million pounds of hazardous waste per year.

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