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Case studies for peptide separation by CPC

Laszlo Lorantfy – CSO – RotaChrom Technologies Ltd.


Centrifugal Partition Chromatography is a liquid-liquid preparative chromatography technique, which efficiently and economically separates compounds without necessitating solid bulk material. In this method, the stationary phase is liquid and is immobilized by a strong centrifugal force. Separation is based on the partition coefficient differential of the compounds of interest and impurities in the mobile and stationary phases. This industrial scale preparative process has multiple applications for various compounds including lipids [1], polyphenols [2] and psychotropic agents [3].

RotaChrom Technologies is currently the exclusive developer and manufacturer of industrial scale CPC instruments (iCPC) capable of processing multiple tons of material per month. This patented system has successfully processed a diverse portfolio of API purification challenges with greater yield, precision, and cost effectiveness than conventional Preparative Liquid Chromatography methods. This list includes, but is not limited to, prostaglandins, antibiotics and oligopeptide separations. Furthermore, in many cases inefficient multi-step crystallization can be effectively replaced by a single-step iCPC separation.

Provided below are two case studies in which RotaChrom drastically increased the yield, reduced the number of steps and decreased costs in peptide drug production through the iCPC process. As a result, the product value as well as profitability of the produced compounds were increased significantly.

Immunosupressant – Case study A

Our Partner faced a serious problem, while increasing production from 3 tons/year to 10 tons/year, for a material costing 1000-1500 Euro/kg. Although they managed to improve upstream to sufficient levels, their purification capacity was not adequate for the level, and some impurities were not manageable with their current liquid chromatographic techniques. This is where we received a small amount of sample to test RotaChrom’s ability to deal with the problem.

Crude sample A

– picture 1.: Crude sample A

Purified material A

– picture 2.: Purified material A

The iCPC system was tested through a feasibility process by which a small amount of material was provided by the company in order to demonstrate proof of concept. The feasibility and subsequent pilot studies replicated the company’s intended Scale up by testing the iCPC method on 1/10,000 to 1/100th the scale of actual production.

The iCPC platform was able to isolate most impurities with a significantly increased yield. The Chromatogram in Picture Two demonstrates the results of the process. Through the iCPC platform, successful separation was achieved without the use of expensive silica gel. With reduced solvent consumption, and the use of only technical grade solvents, the iCPC process was able to reduce the cost of purification by ~30% while achieving impurity levels of only <0.3 % to <0.1 % and an increased production yield of ~20%. Furthermore, iCPC method reduced the number of steps in the purification process from two Prep-LC steps to only one CPC step. Overall it was estimated that the company’s downstream Scale up would have been at least triple without the iCPC system. Finally, the X100 Scale up feasibility simulation had no negative impact on separation parameters including purity and yield.

All indicators demonstrated the superiority of the iCPC method in this downstream purification process. The company decided to adapt this platform in its production chain by Q3 2016. It is estimated that the company will have an increase of 3 million Euro/year with a positive return on investment in approximately one year through the implementation of the system.

The graph below compares cost and benefit of the conventional and iCPC processes:

Cost comparison chart A

Cost comparison chart A

The above graph outlines the costs and “value” associated with the conventional purification method as compared to the iCPC system. The costs of the company’s previous purification system totaled approximately 750 Euros/Kg. Following the transition, the upstream costs of the company were drastically reduced since greater impurity levels were tolerated by the iCPC process. This allowed for raw material costs to decrease by approximately 100 Euros. By only utilizing technical grade solvents, eliminating the cost of silica gel, and marginally increasing the cost of technology, the company saved approximately 250 Euros/kg API. Overall we see an increase in product value (or the value of the product that can be produced from 1 kg of crude) as defined by the following formula:

Product Value = (Product Recovery or Percentage Yield) x (Product market price/kg)

Antibiotic – Case study B

A pharmaceutical company decided to optimize and streamline the production efficiency of an oligopeptide compound. The original manufacturing process consisted of 14 extraction and 2 recrystallization steps for a material costing about 10,000 Euro/kg, and resulted yield of 40-50%. As a generic drug, the company’s aim was to increase the profitability of production.

The iCPC system reduced the original 16 step purification process to a single extraction step utilizing iCPC and final crystallization to achieve the final product. Furthermore, RotaChrom introduced CPC recirculation of the mother liquor resulting in a yield of 80-90%.

The Chromatogram in Picture Three depicts the crude sample provided by the company and the chromatogram in Picture Four demonstrates the results following the single step iCPC process. While the purity level was similar to the conventional 16 step process, the single step iCPC method doubled the product yield.

Crude sample B

– picture 3.: Crude sample B

Purified material B

– picture 4.: Purified material B

The Pharmaceutical Company decided to expand its contract with RotaChrom and is currently working on a 100x scaled-up pilot to streamline the company’s entire manufacturing process through the implementation of iCPC. Based on the data gathered, RotaChrom estimates that the pharmaceutical company will save ~30% of production costs utilizing this method. RotaChrom further estimates that the company will have a positive return on investment in less than two years with an increase of yearly profit of at least 1 million Euros.

The following graph compares cost and benefit of the former and new processes:

Cost comparison chart

Cost comparison chart B

The above graph illustrates how the product value and profitability of the manufacturing process was dramatically increased by replacing inefficient multi-step crystallization and two additional liquid chromatography steps with a single iCPC step.


These case studies demonstrate the radical impact that Industrial Scale CPC will have in optimizing yield, purity, and streamlining production for Oligopeptide manufacturing. As iCPC redefines the standards of this industry, this method will very soon become a mainstream process in purification with the potential to completely replace many conventional Liquid Chromatography systems in the next 5 years.


[1] P. F. Udaya Wanasundara, „Centrifugal Partition Chromatography (CPC): Emerging separation and purification technique for lipid and related compounds,” Food Technology, 1. kötet13, 1. szám1, pp. 726-730, 2002.
[2] C. C. C. C. J. V. Jean-Claude Delaunay, „Preparative isolation of polyphenolic compounds from Vitis vinifera by centrifugal partition chromatography,” Journal of Chromatography A, %1. kötet964,  1. szám1, pp. 123-128, 2002.
[3] K. N. S. S. Osamu Shirota, „Simple preprataive isolation of Salvinorin A from the hallucinogenic sage, Salvia divinorum, by Centrifugal Partition Chromatography,” Journal of Liquid Chromatography & Related Techniques,  1. kötet30,  1. szám1, pp. 1105-1114, 2007.

Case studies for peptide separation by CPC | Rotachrom Technologies LLC.