Pre-column Derivation HPLC Method for Detemination of Aldehydes and Research on the Correlation between Aldehydes and Peroxide Value in Polysorbates / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop a quantitative determination method of formaldehyde and acetaldehyde and study the correlation between aldehydes and peroxide value in polysorbate 20, 40, 60 and 80.METHODS: Formaldehyde and acetaldehyde in polysorbates reacted with 2,4-dinitrophenylhydrazine(DNPH) for 60 min at room temperature and stable phenylhydrazone compounds,formaldehyde-DNPH and acetaldehyde-DNPH, were obtained, respectively.The two phenylhydrazone compounds were simultaneously detected by HPLC to calculate the contents of formaldehyde and acetaldehyde in polysorates. The peroxide value of polysorbates was also determined according to chapter of Chinese Pharmacopoeia 2015. RESULTS: The HPLC method for quantitative determination of aldehydes in polysorates had good specificity. The resolutions of formaldehyde-DNPH and acetaldehyde-DNPH were 2.0 and 2.9, respectively. There was good linearity in the range of 0.1-35.0 μg•mL-1 for formaldehyde(r=0.999 8) and 0.2-50.0 μg•mL-1 for acetaldehyde(r=0.999 6). The average spiked recoveries were in the range of 88.6%-103.5% and their relative standard deviations(RSDs) were in the range of 1.2%-2.6%.The RSDs of intra-day and inter-day precision were not more than 6.0%.The limits of detection and quantitation were 0.03 and 0.1 μg•mL-1 for formaldehyde and 0.06 and 0.2 μg•mL-1 for acetaldehyde, respectively. The contents of aldehydes in polysorbates from different manufacturers were 0-593.1 μg•g-1 for formaldehyde and 0-740.5 μg•g-1 for acetaldehyde, however, the peroxide values of most polysorbate products were less than 10. CONCLUSION: The pre-column derivation HPLC method is suitable for assay of trace aldehydes in polysorbates. Moreover, the weak correlation between aldehydes and peroxide value indicated that the latter testing item is not enough to control the risk of aldehydes in polysorbates. Therefore, aldehydes as well as quantitative determination method are suggested to be included in the monographs of polysorbates to improve their quality standard.