## 2 Materials and Methods
### NVP-13 Concentration Measurement  
Plasma: The analytical work was done in accordance to standard operating procedures and for each chromatographic analysis batch, a detailed assay protocol was generated and retained as raw data. The method was validated at Axolabs prior to the start of this study phase. Results of the method validation are summarized in the Method Validation Report for the PNA-HPLC-Assay for cynomolgus monkey plasma: AN57.0-17. The analytical method is based on an AEX-HPLC method with fluorescence detection that allows the sensitive and specific detection of NVP-13 from cynomolgus monkey plasma samples. The assay is based on the specific hybridization of a 16-mer complementary PNA-probe conjugated at the N-terminus with Atto425 dye. The duplex of PNA and parent compound yields a specific signal in the subsequent analysis by AEX-HPLC coupled to a fluorescence detector, as described in detail earlier [21]. Quantification is performed based on an external calibration curve generated from a standard dilution series in cynomolgus monkey plasma. Linear calibration curves (weighted 1/X) are calculated from 1 ng/mL to 5000 ng/mL. CSF: The method of analysis used in this study is documented in Standard test Method: STM_NVP-13_CynoCSF. The analytical work was done in accordance with Axolabs' SOP \begin{table}
\begin{tabular}{c c c} \hline
**Code** & **Parameter** & **Method of Determination** \\ \hline AST. & Aspartate Aminotransferase & IFCC Reference Method with Pyridoxal \\  & & Phosphate Activation \\ ALT. & Alanine Aminotransferase & IFCC Reference Method with Pyridoxal \\  & & Phosphate Activation \\ ALP. & Alkaline Phosphatase & Colorimetric Method using P Nitrogenly \\  & & Phosphate as Substrate Standardized to IFCC \\ CHOL. & Total Cholesterol & Enzymatic Method using Cholesterol \\  & & Oxidase/Esterase \\ TRIG. & Triglycerides & Enzymatic Method using Lipase and Glycerol \\  & & Kinase as Primary Enzymes \\  & & A colorimetric Method (Malloy-Evelyn) in which \\  & & indirect Bilirubin is liberated by Detergent. Total \\ TBIL. & Total Bilirubin & Bilirubin is then coupled with a Diazonium \\  & & Compound to give \\  & & Corresponding Azobilirubin. \\ TP. & Total Protein & Biuret Method \\ ALB. & Albumin & Colorimetric Method using Bromocresol Green \\  & & Calculation (Globulin = Total \\ GLOB. & Globulin & protein – Albumin) \\ A:G. & Albumin/Globulin Ratio & Calculation (Albumin/Globulin) \\ NA. & Sodium & Ion Selective Electrode (Direct Potentiometric) \\ K. & Potassium & Ion Selective Electrode (Direct Potentiometric) \\ CL. & Chloride & Ion Selective Electrode (Direct Potentiometric) \\ CA. & Calcium & Colorimetric Method using Arsenazo III \\ PHOS. & Inorganic Phosphorus & UV Method using Ammonium Molybdate \\ CREA. & Creatinine & Colorimetric Method using Piciric Acid (Standard \\  & & Jaffe Method) \\ BU. & Blood Urea & UV Method using a coupled Urease Procedure \\ GLDH. & Glutamate Dehydrogenase & UV Method with optimized Substrate \\  & & Concentration (Optimized DGKC Method) \\ GGT. & Gamma Glutamyltransferase & Standardized IFCC colorimetric Method \\ GLU. & Glucose & UV Method using a Coupled \\  & & Hexokinase Procedure \\ \hline \end{tabular}
\end{table} 
Table 2: Summary of clinical chemistry parameters investigated throughout every phase throughout the experiment. RA-001 and for each chromatographic analysis batch, a detailed assay protocol was generated and retained as raw data. The method was validated at Axolabs prior to the start of this study phase. Results of the method validation are summarized in the Method Validation Report for the PNA-HPLC-Assay for cynomolgus monkey plasma: AN58.0-17. The analytical method is based on an AEX-HPLC method as described above (Plasma section) using cynomolgus CSF instead of plasma for calibration. Tissue: The method of analysis used is documented in the Method Development Report: MDR NVP-13 & 19_PNA-HPLC-Assay.