Volume 5 Supplement 8

22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies

Open Access

Analytical techniques for characterization of raw materials in cell culture media

  • Chandana Sharma1,
  • Barry Drew1,
  • Kevin Head1,
  • Rani Pusuluri1 and
  • Matthew V Caple1
BMC Proceedings20115(Suppl 8):P5

DOI: 10.1186/1753-6561-5-S8-P5

Published: 22 November 2011

Abstract

Raw materials are a critical part of any cell culture medium; therefore, it is of utmost importance to understand and characterize them for high-quality product. The raw material characterization (RMC) program at SAFC focuses on individual screening of raw materials both analytically and biologically. The goal of the program is to develop the best-in-class knowledge base of the raw materials used in SAFC’s media formulations and their impact on performance of products.

Background

A prioritized list of 100 “high-risk” raw materials was developed based on a risk assessment performed within SAFC. This poster will focus on the analytical screening of certain “high-risk” raw materials within the prioritized list to identify any variability and critical contaminants present. In order to achieve this, orthogonal methods were used that include ultra-high performance liquid chromatography-mass spectrometry (U-HPLC/MS) for non-volatile polar components and gas chromatography-mass spectrometry (GC/MS) for volatile non-polar materials. Inductively coupled plasma-optical emission spectrometry (ICP-OES) was also used to identify any trace metal contamination present. In addition, the solubility of the raw materials is also tested to identify any variability within a vendor or between different vendors.

Results

Figure 1 shows the process flow followed within the analytical RMC initiative. It describes how each raw material is screened and the strategy for analyzing any possible contaminant.
https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-5-S8-P5/MediaObjects/12919_2011_Article_923_Fig1_HTML.jpg
Figure 1

Analytical RMC process flow

Table 1

Summary of solubility findings on multiple lots of three amino acids.

 

Solubility in 100 ml of a neutral buffer

Vendor

Lot No.

Lysine.HCl (g)

Cystine.2HCl (mg)

Tyrosine.2Na (mg)

A

1

5.0

12.4

175.1

 

2

12.5

35.0

185.8

 

3

10.0

35.2

185.5

B

1

15.0

30.3

185.8

 

2

12.5

50.8

195.2

 

3

12.0

48.2

200.7

C

1

60.0

32.0

N/A

 

2

60.0

35.2

N/A

 

3

60.0

32.8

N/A

D

1

60.0

N/A

N/A

 

2

60.0

N/A

N/A

Conclusions

The orthogonal methods cited to characterize raw materials have proven to be robust and reliable for the intended purpose. The solubility experiment described for amino acids illustrates a significant difference in solubility limits of amino acids and establishes intra- and inter-vendor variability. This program has helped SAFC get better insight into their suppliers’ manufacturing processes. This is a long term initiative within the organization and the most important goal through the program is to develop a better understanding of raw materials to deliver superior products of the finest quality.

Declarations

Acknowledgements

Our sincere thanks to Mr. T. Bower and Mr. S. Jones of Sigma-Aldrich for running samples on ICP and on LC/MS respectively.

Authors’ Affiliations

(1)
Cell Sciences and Development, SAFC

Copyright

© Sharma et al; licensee BioMed Central Ltd. 2011

This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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