The use of ultrafiltration for removing interferences in the Lowry microassay of extractable proteins in natural rubber latex gloves

Esah Y., . and Mok K. L., . and Lai P. F., . The use of ultrafiltration for removing interferences in the Lowry microassay of extractable proteins in natural rubber latex gloves. pp. 17-27. ISSN 1511-1768

This study evaluates the use of ultrafiltration in removing micro-solutes that might interfere in the Lowry microassay. Reference proteins glove extracts and 1 3-diphenylguanidine were subjected to ultrafiltration using a commercial concentrator which has a membrane filter of 3 kD molecular weight cutoff. Separated fractions were analysed for their reactivity by the MS 1392: 1998 Lowry mircoassay. Purified reference proteins with molecular weights greater than 3 kD showed high retentate recovery which verified the efficacy of separation of the ultrafiltration unit. Micro-solutes were evidently present in both the non-interfering and interfering glove extracts. Irrespective of their original extractable protein (EP) values the micro-solute content for the non-interfering glove extracts was relatively constant at 28. This is in contrast with the marked variation observed for the interfering glove extracts. The ultrafiltration process caused an irrecoverable loss of approximately 5 - 6. The total recovery was comparable for both the non-interfering and interfering glove samples. This indicates that the micro-solutes and/or their interfering components were not detrimental to the filter membrane causing no obvious degradation of membrane integrity. Some samples may however necessitate higher number of wash cycles to completely remove the interference and obtain accurate EP values.

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Abstract

This study evaluates the use of ultrafiltration in removing micro-solutes that might interfere in the Lowry microassay. Reference proteins glove extracts and 1 3-diphenylguanidine were subjected to ultrafiltration using a commercial concentrator which has a membrane filter of 3 kD molecular weight cutoff. Separated fractions were analysed for their reactivity by the MS 1392: 1998 Lowry mircoassay. Purified reference proteins with molecular weights greater than 3 kD showed high retentate recovery which verified the efficacy of separation of the ultrafiltration unit. Micro-solutes were evidently present in both the non-interfering and interfering glove extracts. Irrespective of their original extractable protein (EP) values the micro-solute content for the non-interfering glove extracts was relatively constant at 28. This is in contrast with the marked variation observed for the interfering glove extracts. The ultrafiltration process caused an irrecoverable loss of approximately 5 - 6. The total recovery was comparable for both the non-interfering and interfering glove samples. This indicates that the micro-solutes and/or their interfering components were not detrimental to the filter membrane causing no obvious degradation of membrane integrity. Some samples may however necessitate higher number of wash cycles to completely remove the interference and obtain accurate EP values.

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Item Type:	Article
AGROVOC Term:	Natural rubber
AGROVOC Term:	Molecular weight
AGROVOC Term:	Purification
AGROVOC Term:	Ultrafiltration
AGROVOC Term:	Proteins
AGROVOC Term:	Latex
AGROVOC Term:	Solutes
AGROVOC Term:	Separation
AGROVOC Term:	Degradation
AGROVOC Term:	Membranes
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 06:28
URI:	http://webagris.upm.edu.my/id/eprint/23185

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