Objective: To establish an HPLC-Q-Orbitrap method to determine the contents of genotoxic impurities methyl isoquinoline-5-sulfonate and ethyl isoquinoline-5-sulfonate in fasudil hydrochloride. Methods: The separation was performed on a Thermo Scientific Hypersil GOLD C8 column（4.6 mm×150 mm,3 μm）using the mobile phase consisting of acetonitrile（mobile phase A）and 0.1% formic acid aqueous solution（mobile phase B）by gradient elution at a flow rate of 0.9 mL·min-1．The column temperature was set at 30 ℃．The analytes were detected by Q-Orbitrap high resolution mass spectrometry with ESI source in positive mode．The detection was achieved in Full MS-SIM mode,and the scan range was set at m/z 223-239．The extracted ion chromatograms were used for the quantitation,selecting m/z 224.037 59（mass tolerance<5×10-6）and m/z 238.053 24（mass tolerance<5×10-6）as the quantitative ions of methyl isoquinoline-5-sulfonate and ethyl isoquinoline-5-sulfonate, respectively. Results: Methyl isoquinoline-5-sulfonate and ethyl isoquinoline-5-sulfonate showed good linear relationships within the range of 1.0-30 ng·mL-1（r=0.998 3 and 0.999 0,respectively）．The limit of detection and limit of quantification for methyl isoquinoline-5-sulfonate was 0.66 μg·g-1 and 2.0 μg·g-1,respectively．The limit of detection and limit of quantification for ethyl isoquinoline-5-sulfonate was 0.30 μg·g-1 and 1.0 μg·g-1,respectively． The spike recoveries（n=6）of the two compounds were 95.4% and 97.1%,with RSD of 2.1% and 1.9%, respectively．The two genotoxic impurities were not detected in 6 batches of samples. Conclusion: The established method is simple,rapid,accurate and sensitive,and can be used for the quality control of fasudil hydrochloride.
LIU Yang, LI Jian-feng, LI Ning, CHEN Yun-jian, LIU Shi-cheng, YANG Zhao-xiang
. Determination of genotoxic impurities methyl isoquinoline-5-sulfonate and ethyl isoquinoline-5-sulfonate in fasudil hydrochloride by Q-Orbitrap high resolution mass spectrometry*[J]. Chinese Journal of Pharmaceutical Analysis, 2021
, 41(1)
: 154
-159
.
DOI: 10.16155/j.0254-1793.2021.01.19
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