Advancements in Analyzing Tumor Metabolites through Chemical Derivatization-Based Chromatography.

Understanding the metabolic abnormalities of tumors is crucial for early diagnosis, prognosis, and treatment. Accurate identification and quantification of metabolites in biological samples are essential to investigate the relationship between metabolite variations and tumor development. Common techniques like LC-MS and GC-MS face challenges in measuring aberrant metabolites in tumors due to their strong polarity, isomerism, or low ionization efficiency during MS detection. Chemical derivatization of metabolites offers an effective solution to overcome these challenges. This review focuses on the difficulties encountered in analyzing aberrant metabolites in tumors, the principles behind chemical derivatization methods, and the advancements in analyzing tumor metabolites using derivatization-based chromatography. It serves as a comprehensive reference for understanding the analysis and detection of tumor metabolites, particularly those that are highly polar and exhibit low ionization efficiency.

Thalassotalea algicola sp. nov., an alginate-utilizing bacterium isolated from a red alga.

A facultatively anaerobic bacterium, strain M1531T, was isolated from a red alga (Porphyra) at coastal water in Weihai, China. Cells of the novel strain were Gram-stain-negative, rod-shaped, motile by means of a single polar flagellum and around 0.6-0.8 × 2.0-3.0 µm in size. Optimum growth occurred at 30 °C, with 2% (w/v) NaCl and at pH 6.5-7.0. On the basis of the result of phylogenetic analysis of the 16S rRNA gene sequence, stain M1531T had close relative with Thalassotalea euphylliae KCTC 42743T (96.9%). Genome sequencing revealed a genome size of 4,061,950 bp, a G + C content of 39.1 mol% and four protein-coding genes related to the degradation of alginate. According to the data obtained, strain M1531T shared ANI value below 95-96%, dDDH value below 23.8% with the closely related type species. Strain M1531T had Q-8 as the predominant isoprenoid quinone and possessed Summed Features 3 (C16:1 ω7c/C16:1 ω6c), C16:0 and Summed Features 8 (C18:1 ω7c/C18:1 ω6c) as the major fatty acids. The polar lipids of strain M1531T were identified as phosphatidylglycerol, phosphatidylethanolamine, one unidentified phospholipid, one unidentified aminolipid and four unidentified lipids. According to the results of the phenotypic, chemotaxonomic characterization, phylogenetic properties and genome analysis, strain M1531T represents a novel specie of the genus Thalassotalea, for which the name Thalassotalea algicola sp. nov. is proposed. The type strain is M1531T (= MCCC 1H00400T = KCTC 72865T).

[Determination of dacarbazine in the urine of mice with melanoma by high performance liquid chromatography].

Dacarbazine (DTIC) is a first-line chemotherapy drug that is widely used in clinical practice for malignant melanoma. DTIC is metabolized by the liver in vivo. Some drugs are excreted in urine in the form of a prototype. Hence, DTIC in urine can be monitored to evaluate its utilization and conversion rate in the human body, and then to determine its therapeutic effect. Urine is the only body fluid that can be obtained in large quantities without damage, and it plays an important role in the analysis of body functions. However, the composition of urine is complex and there is large matrix interference, because of which trace analysis or trace component analysis is difficult. At present, the main analytical methods for DTIC are high performance liquid chromatography (HPLC) with/without mass spectrometry (MS). HPLC and HPLC-MS have the advantages of good separation effect, good selectivity, high detection sensitivity, automatic operation, and wide application range. Unfortunately, DTIC is a strongly polar and weakly basic compound; thus, it is difficult to achieve good separation and obtain good peak shapes by conventional reversed-phase chromatography. To overcome these defects, it is necessary to develop a novel method for the analysis of DTIC. In this study, mice were subjected to 12 h of fasting; then, blueberry anthocyanin was administered by gavage, and DTIC was administered by intraperitoneal injection. Then, morning urine was collected in a metabolic cage. Urine collection was continued every 4 days for a total of 5 times. Within 2 h, the collected urine was centrifuged (3000 g, 4℃) for 10 min to remove solids. The supernatant was stored in a refrigerator at-80℃. Before analysis, the urine samples were removed from the refrigerator and thawed naturally at room temperature. Then, the samples were treated by the acetone-sediment method, freeze-dried, dissolved in the mobile phase, and subjected to HPLC analysis with isocratic elution. The separation was performed on a Shimadzu-GL ODS column (250 mm×4.6 mm, 5 µm). The mobile phase was methanol/acetonitrile (1:1, v/v)-0.01 mol/L NaH2PO4 (pH 6.5; 20:80, v/v) at a flow rate of 1 mL/min. The detection wavelength, column temperature, and running time were 280 nm, 30℃, and 15 min, respectively. Under the optimized conditions, the retention time of DTIC was 5.3 min, and a good peak shape was obtained. The linearity ranged from 0.25 to 1000 µg/mL (r2=0.999). The limits of detection and quantification were calculated to be 0.12 µg/mL and 0.25 µg/mL based on signal-to-noise ratios of 3 and 10, respectively. At three spiked levels (50.0, 375, and 500 µg/mL), the average recoveries were 98.9%, 102%, and 99.1% with relative standard deviations (RSDs) of 3.2%, 1.3%, and 1.2% (n=5), respectively. The RSDs of the interday and intraday measurements were lower than 3.8% and 4.4%, respectively. The proposed method allowed for the accurate determination of DTIC in urine using a mixed organic solvent/phosphate buffer solution as the mobile phase, with equivalent elution for 15 min. This method was successfully applied to monitor the change in DTIC concentration in the urine of C57BL/6 mice in various stages of melanoma. The results demonstrate that the method is simple, reliable, and easy to apply.