Polyunsaturated Fatty Acid-Derived Lipid Mediators as Potential Biomarkers for Leprosy Among Individuals with Asymptomatic Mycobacterium leprae Infection.

Intra-household contacts (HCs) of leprosy patients are at increased risk of infection by Mycobacterium leprae and about ∼5-10% will develop active disease. A prognostic tool to identify HCs with the greatest risk of progressing to active disease would enhance early leprosy diagnosis and optimize prophylactic intervention. Previous metabolomics studies suggest that host lipid mediators derived from ω-3 and ω-6 polyunsaturated fatty acids (PUFAs) are potential biomarkers for leprosy. In this study, we investigated retrospective sera of leprosy HCs by liquid chromatography-mass spectrometry and enzyme-linked immunoassay to determine whether circulating levels of ω-3 and ω-6 PUFA metabolites were altered in HCs that developed leprosy (HCDL) in comparison to those that did not (HCNDL). Sera were collected from HCs at the time of index case diagnosis and before clinical signs/symptoms of leprosy. Our findings showed that HCDL sera exhibited a distinct metabolic profile in comparison to HCDNL. Specifically, arachidonic acid, leukotriene B4, 11-hydroxyeicosatetraenoic acid, prostaglandin D2, and lipoxin A4 were elevated in HCDL. In contrast, prostaglandin E2 levels were reduced in HCDL. The ω-3 PUFAs, docosahexaenoic acid, eicosapentaenoic acid, and the docosahexaenoic acid-derived resolvin D1 and maresin-1 were also elevated in HCDL individuals compared to HCNDL. Principal component analyses provided further evidence that lipid mediators could serve as an early biomarker for progression to active leprosy. A logistic model identified resolvin D1 and D2, and prostaglandin D2 as having the greatest potential for early detection of HCs that will manifest leprosy.

High-performance liquid chromatography-based multivariate analysis to predict the estrogenic activity of an Epimedium koreanum extract.

This study characterizes the correlation between the chemical fingerprint and estrogenic activity of an Epimedium koreanum extract. The estrogenic activity of 31 E. koreanum extract samples was evaluated by a luciferase reporter gene assay, and the samples were classified into 3 groups based on their bioactivity. A chemical fingerprint analysis was performed on each sample by high-performance liquid chromatography (HPLC), and 44 common peaks were selected from the chromatogram and used as a dataset for a pattern recognition analysis. A canonical discriminant analysis performed on this dataset determined a distinct distribution of the samples according to their estrogenic activity on the scoring plot. The classification results showed that 90.3% of the original grouped cases had been correctly classified. The total content of the 4 major extract compounds, epimedin A, epimedin B, epimedin C, and icariin, exhibited good correlation (r=0.784) with the estrogenic activities of the respective extracts. This chromatographic fingerprint-chemometric analysis system could be useful for predicting the E. koreanum pharmacological activity and consequent biological activity-relevant quality control assessment.

A simple isocratic HPLC method for the simultaneous determination of bioactive components of Scutellariae radix extract.

Scutellariae Radix, the dried root of Scutellaria baicalensis Georgi, has been widely used in Asian countries for the treatment of dermatitis, diarrhoea, inflammatory disease and hepatic disease. A simple, sensitive and precise reversed-phase liquid chromatographic method with isocratic elution was developed to simultaneously determine four bioactive compounds in Scutellariae Radix: baicalein, baicalin, wogonin and wogonoside. Chromatographic analysis was performed on a YMC Pack Pro C(8) column (150 × 4.6 mm(2), 3 µm), with a mobile phase of 0.1% formic acid : acetonitrile (70 : 30, v/v) at a flow rate of 1.0 mL min(-1), and UV detection at 280 nm. Linear behaviour was observed over the investigated concentration range (0.25-10 µg mL(-1)) for all analytes, with a correlation coefficient of >0.997. The intra- and inter-day precisions were <8.07%, and accuracies were 92.3-102.9%. This method was successfully applied for the analysis of marker compounds for the quality control of Scutellariae Radix extract.

Simultaneous determination of phenolic acids and phthalide compounds by liquid chromatography for quality assessment of Rhizoma cnidii.

This paper describes a simple, rapid, and validated reversed-phase high-performance liquid chromatographic method developed for the determination of 4 major bioactive constituents, namely, chlorogenic acid, ferulic acid, senkyunolide A, and Z-ligustilide in Rhizoma Cnidii extract. A Capcell Pak C18 chromatographic column (150 x 4.6 mm, 3 microm) was used with mobile phases consisting of 0.1% formic acid, acetonitrile, and methanol at a flow rate of 0.8 mL/min and UV detection at 285 nm. Comprehensive validation of the method included evaluation of linearity, repeatability, recovery, and stability. Excellent linear behavior (r2>0.99) was observed over the concentration range of 2-100 microg/mL for the compounds under investigation. Repeatability and accuracy were evaluated by intra- and interday assays; the relative standard deviation (RSD) values were < or = 5.37% and accuracies ranged from 97.1 to 104.9%. Recoveries of the compounds ranged from 94.2 to 104.2% with RSD values of < or = 9.50%. The developed method was successfully applied to the analysis of ethanolic extracts of Rhizoma Cnidii samples. As a result, the concentrations of chlorogenic acid, ferulic acid, Z-ligustilide, and senkyunolide A were determined to be 0.84-5.35, 0.45-1.65, 0.74-4.39, and 0.32-1.14 mg/g herb, respectively. Thus, the developed method was found to be accurate and reproducible and is considered suitable for the qualitative and quantitative analysis of Rhizoma Cnidii for bioactive compounds.

Simultaneous determination of bioactive xanthone glycosides and norlignans from ethanolic extract of Anemarrhena asphodeloides by liquid chromatography.

The rhizomes of Anemarrhena asphodeloides Bunge (Liliaceae) are prescribed as crude drugs in herbal medication for the treatment of various diseases such as diabetes, inflammation, and platelet aggregation inhibition. A simple, sensitive, and precise reversed-phase liquid chromatographic method was developed to study the quantitative determination of 5 bioactive compounds from these rhizomes, namely, neomangiferin, mangiferin, isomangiferin, nyasol, and methylnyasol. Chromatographic analysis was performed on Capcell Pak C18 column (150 x 4.6 mm, 3 microm) with a mobile phase consisting of acetonitrile, methanol, and 0.1% formic acid at a flow rate of 1.00 mL/min. Quantitation was performed using a UV-visible detector at 260 nm. The method for the determination of reported medicinal agents was accurate and reproducible. Excellent linear behavior was observed over the investigated concentration range of 2.5-100.0 microg/mL for neomangiferin; 1.5-60.0 microg/mL for mangiferin; 0.5-20.0 microg/mL for nyasol; and 0.2-20.0 microg/mL for methylnyasol; correlation coefficient > 0.99. The intraday and interday precision over the concentration range of compounds was < 6.6% (relative standard deviation) and accuracy was between 94.9 and 109.3%. This method can be successfully applied for the analysis of medicinal compounds from the ethanolic extract of A. asphodeloides Bunge.