HPLC-UV Estimation of Folic Acid in Fortified Rice and Wheat Flour using Enzymatic Extraction and Immunoaffinity Chromatography Enrichment: An Interlaboratory Validation Study.

BACKGROUND: In India, fortification of cereals with folic acid has been voluntary for many years. However, The Food Safety and Standards Authority of India's recent Fortification of Foods Regulations (2018) has prompted us to develop and validate a simple analytical method for estimation of folic acid in fortified cereals. OBJECTIVE: The aim was to develop and validate a simple and rugged HPLC-UV method for quantitative analysis of folic acid in fortified rice and wheat flour. METHODS: The enzymatic sample extract was diluted with phosphate buffer, centrifuged, filtered, and then passed slowly through an immunoaffinity cartridge for cleanup. Folic acid in the sample extract was retained by the cartridge and subsequently eluted with 30% acetonitrile [+0.2% trifluoroacetic acid (TFA)]. The elute was collected and analyzed by HPLC-UV at 280 nm. The chromatographic separation of folic acid was achieved on an Agilent Poroshell SB-C18 column (3.0 × 100 mm, 2.7 µm) with 0.1% TFA in methanol as mobile phase. RESULTS: The linearity range of the vitamin was established in the concentration range of 50-800 µg/L, and the regression coefficient was more than 0.999. The LOQ was 5 µg/L. The average spike recovery values of folic acid in rice and wheat flour samples were 90.9 and 80.5%, respectively. The method was subjected to an interlaboratory validation; eight accredited food testing laboratories across India participated in it and resulted in satisfactory z-scores for the reported results. CONCLUSIONS: The method will be useful in regulatory compliance testing of folic acid in fortified cereals and processed products. HIGHLIGHTS: A sensitive analysis method is reported for estimation of folic acid in fortified rice and wheat flour. The scope, selectivity, repeatability, and reproducibility of the method establishes it as fit for regulatory compliance check purposes.

Discovery of pyrazole carboxylic acids as potent inhibitors of rat long chain L-2-hydroxy acid oxidase.

Long chain L-2-hydroxy acid oxidase 2 (Hao2) is a peroxisomal enzyme expressed in the kidney and the liver. Hao2 was identified as a candidate gene for blood pressure (BP) quantitative trait locus (QTL) but the identity of its physiological substrate and its role in vivo remains largely unknown. To define a pharmacological role of this gene product, we report the development of selective inhibitors of Hao2. We identified pyrazole carboxylic acid hits 1 and 2 from screening of a compound library. Lead optimization of these hits led to the discovery of 15-XV and 15-XXXII as potent and selective inhibitors of rat Hao2. This report details the structure activity relationship of the pyrazole carboxylic acids as specific inhibitors of Hao2.

Development and validation of a stability indicating method for the enantioselective estimation of omeprazole enantiomers in the enteric-coated formulations by high-performance liquid chromatography.

Omeprazole is widely prescribed in the form of enteric-coated formulations, due to the rapid degradation of the drug in the acidic condition of the stomach. In the current article, we are reporting the development and complete validation of a stability indicating chiral high-performance liquid chromatography (HPLC) method for the enantioselective analysis of omeprazole in the enteric-coated formulations. A precise and sensitive enantiomeric separation of omeprazole was obtained on Chiralcel OD-H analytical column (250mm × 4.6 mm, 5µm particle size) using normal phase chromatography. The analysis was performed under UV detection at 301nm wavelength. During method development, the addition of methanol to the mobile phase helped in getting the sharp peaks. The developed method showed linear response over a wide concentration range of 0.39-800µg/ml and the regression coefficients value (r(2)) was obtained more than 0.999 for (S)- and (R)-omeprazole. The lower limit of detection (LLOD) and lower limit of quantification (LLOQ) for (R)-omeprazole were found to be 0.39 and 0.78 µg/ml, respectively for 5 µl injection volume. The percentage recovery of (R)-omeprazole ranged from 93.5 to 104 in spiked formulation samples and omeprazole sample solution and mobile phase were found to be stable for at least 24 h at room temperature. The proposed method was found to be suitable and accurate for the quantitative determination of undesired enantiomer in the enteric-coated omeprazole formulations.

Impact of a clinical nurse specialist on the treatment of chronic hepatitis C.

Hepatitis C virus is a common cause of liver disease and a major health problem worldwide. Modern treatment strategies can be successful in up to 50% of cases, with patients treated in specialist and more general hospital settings. Guidelines issued by the Department of Health and the then National Institute for Health and Clinical Excellence made it obligatory for a clinical nurse specialist (CNS) to be involved with assessing and treating patients with chronic hepatitis C. The aim of this retrospective case study was to assess the impact on referral, assessment and treatment of patients with chronic hepatitis C before and after the introduction of a CNS to Salisbury District Hospital. There was a significant improvement in patient follow-up and intention to treat, with a doubling in treatment rates. This was achieved by improving clinic attendance after the patient's initial outpatient assessment, allowing improved biopsy rates and subsequent selection for antiviral therapy. However, over half the cases with chronic hepatitis C identified in primary care each year were not referred to secondary care for assessment.