Generic MS-based method for the bioanalysis of therapeutic monoclonal antibodies in nonclinical studies.

Aim: A generic bioanalytical method was developed to quantify therapeutic IgG1 monoclonal antibodies (mAbs) in mouse sera by combining an easy sample preparation method with LC/MS using selected reaction monitoring. Materials & methods: Rituximab and trastuzumab were used as model mAbs. A synthetic stable isotope-labeled peptide or a stable isotope-labeled mAb was used as an internal standard. The method feasibility was evaluated by a collaborative study involving six laboratories. Results: The calibration curve ranged from 1.0 to 1000.0 µg/ml (correlation coefficient >0.99). The validation parameters including selectivity, linearity of calibration curve, accuracy and precision met the predefined acceptance criteria. Conclusion: Our method is a useful bioanalytical method for the quantification of therapeutic IgG mAbs in nonclinical animal studies.

Proposed selection strategy of surrogate matrix to quantify endogenous substances by Japan Bioanalysis Forum DG2015-15.

It is important to select an appropriate surrogate matrix for preparing calibration standards and quality control samples while quantitatively assaying for endogenous substances, because a blank matrix that does not contain the endogenous substance cannot be derived from the species from which the target study samples are collected. This is because the assay results might be affected, depending on the characteristics of the analyte in the surrogate matrix. Our discussion group that participated in the Japan Bioanalysis Forum discussed the recommended selection strategies, focusing on large and small molecules in ligand binding assays and LC-MS, respectively. We established an efficient selection strategy for a surrogate matrix, with simple compositions as the first candidates stated in this article.

[Effect of pH and Additives on the Compatibility between Vancomycin and Furosemide Injections].

　The physicochemical compatibility between injections of different agents is very important. An injection of the antibiotic vancomycin (VCM) is acidic and its standard pH range is 2.5-4.5. In clinical treatments, VCM injections are often used with Lasix® (furosemide) injections. The Lasix® injection is alkaline and its standard pH range is 8.6-9.6. Therefore, mixing VCM injections with Lasix® injections may cause compatibility problems. We evaluated the effect of pH on the compatibility between VCM (original and two generic) and Lasix® injections. Compatibility was not observed in non-pH-adjusted VCM with Lasix® injections, but white crystals appeared when VCM injections adjusted to pH 2.5 experimentally were mixed with a Lasix® injection, suggesting that the acidic condition of VCM injections cause compatibility. However, the residual rates of VCM did not change after 24 h in all mixtures. We analyzed the crystals by mass spectrometry and 1H-NMR, and identified them to comprise furosemide.

Linoleic acid 10-hydroperoxide as an intermediate during formation of 1-octen-3-ol from linoleic acid in Lentinus decadetes.

In order to confirm the biosynthetic pathway to 1-octen-3-ol from linoleic acid, a crude enzyme solution was prepared from the edible mushroom, Lentinus decadetes. When the reaction was performed in the presence of glutathione peroxidase, which can reduce organic hydroperoxide to the corresponding hydroxide, the amount of 1-octen-3-ol formed from linoleic acid was decreased. At the same time, an accumulation of linoleic acid 10-hydroxide could be detected. The 10-hydroperoxide therefore seems to be an intermediate on the biosynthetic pathway.