Розробка спектрофотометричної методики визначення розувастатину в таблетках з використанням бромфенолового синього

The aim of the work was to develop a spectrophotometric method for the determination of rosuvastatin in tablets based on the reaction with BPB in compliance with the principles of «green» chemistry. Material and methods. Analytical equipment: two-beam UV-visible spectrophotometer Shimadzu model -UV 1800 (Japan), software UV-Probe 2.62, electronic laboratory balance RAD WAG AS 200/C. The following APIs, dosage forms, reagents and solvents were used in work: pharmacopoeial standard sample (CRS) of rosuvastatin calcium (Sigma-Aldrich, (≥ 98 %, HPLC)), BCG (Sigma-Aldrich, (≥ 98 %, HPLC)), "Rosuvastatin" tablets 10 mg, 15 mg, 20 mg, methanol (Honeywell, (≥ 99.9 %, GC)), ethanol (Honeywell, (≥ 99.9 %, GC)), chloroform (Honeywell, (≥ 99.9 %, GC)), acetonitrile (Honeywell, (≥ 99.9 %, GC)), and ethyl acetate (Honeywell, (≥ 99.7 %, GC)). Results and discussion. A spectrophotometric method for determining rosuvastatin by reaction with BPB in an acetonitrile solution using the absorption maximum at a wavelength of 595 nm has been developed. Stoichiometric ratios of reactive components were established, which were 1:1. The developed method for the quantitative determination of rosuvastatin was validated following the requirements of the SPhU. The analytical method was linear in the 7.99-23.97 μmol/L concentration range. The LOD and LOQ values were calculated to be 0.77 µmol/L and 2.36 µmol/L. According to the «greenness» pictogram of the analytical method using the AGREE method, the score was 0.77, indicating that the proposed spectrophotometric method for determining rosuvastatin was developed in compliance with the principles of «green» chemistry. Conclusions. An eco-friendly spectrophotometric method has been developed to quantitatively determine rosuvastatin in tablets based on the reaction with BPB. The appropriate sulfophthalein dye (BPB) and its concentration (4.00 x 10-4), the optimal eco-friendly solvent (acetonitrile), and the appropriate wavelength (595 nm) were chosen, and the sensitivity of the reaction was calculated. The analytical method was validated, and its possibility for use in the pharmaceutical analysis was shownМетою роботи було розробити спектрофотометричну методику визначення розувастатину в таблетках на основі реакції з БФС відповідно до принципів «зеленої» хімії. Матеріали та методи. Аналітичне обладнання: двопроменевий УФ-видимий спектрофотометр Shimadzu model -UV 1800 (Японія), програмне забезпечення UV-Probe 2.62, ваги лабораторні електронні RAD WAG AS 200C. У роботі використовували наступні АФІ, лікарські форми, реактиви та розчинники: фармакопейний стандартний зразок (ФСЗ) розувастатину кальцію (Sigma-Aldrich, (≥ 98 %, ВЕРХ)), БФС (Sigma-Aldrich, (≥ 98 %). ВЕРХ)), «Розувастатин» таблетки 10 мг, 15 мг, 20 мг, метанол (Honeywell, (≥ 99,9 %, ГХ)), етанол (Honeywell, (≥ 99,9 %, ГХ)), хлороформ (Honeywell, (≥ 99,9) %, ГХ)), ацетонітрил (Honeywell, (≥ 99,9 %, ГХ)) та етилацетат (Honeywell, (≥ 99,7 %, ГХ)). Результати та обговорення. Розроблено спектрофотометричну методику визначення розувастатину за реакцією з БФС у розчині ацетонітрилу з використання максимуму поглинання за довжини хвилі 595 нм. Встановлено стехіометричні співвідношення реакційноздатних компонентів, які становили 11. Розроблена методика кількісного визначення розувастатину була валідована відповідно до вимог ДФУ. Аналітична методика була лінійною в діапазоні концентрацій 7,99-23,97 мкмоль/л. Розраховані значення МВ та МКВ становили 0,77 мкмольл і 2,36 мкмольл. Відповідно до піктограми «зеленості» аналітичної методики за методом AGREE, бал складав 0,77 і вказує на те, що запропоновану спектрофотометричну методику визначення розувастатину розроблено з дотриманням принципів «зеленої» хімії. Висновки. Розроблено екологічно чисту спектрофотометричну методику кількісного визначення розувастатину в таблетках на основі реакції з БФС. Обрано відповідний сульфофталеїновий барвник (БФС) і його концентрація (4,00 x 10-4), оптимальний екологічно чистий розчинник (ацетонітрил), відповідну довжину хвилі (595 нм) та розрахована чутливість реакції. Проведено валідацію аналітичної методики та показано її можливість використання у фармацевтичному аналіз

Nitric oxide-mediated effects of L-ornithine-L-aspartate in acute toxic liver injury

This study was aimed to investigate nitric oxide-dependent mechanisms of L-ornithine-L-aspartate (LOLA) action in acute toxic liver injury in rats. Acute hepatitis was induced in Wistar rats using 50% oil solution of tetrachloromethane (CCl4) intragastrically (2 g/kg) twice in a 24 hour interval. Intraperitoneal treatment with LOLA (200 mg/kg) was started 6 hours after the second CCl4 administration and maintained for 3 consecutive days. L-Nω-Nitroarginine Methyl Ester (L-NAME) was used intraperitoneally (10 mg/kg). In CCl4-induced hepatitis, LOLA restores the structure of hepatocytes and prevents aminotransferases, alkaline phosphatase and gamma-glutamyl transferase elevation. It decreases total bilirubin concentration but does not affect increased cholesterol level. LOLA augments urea concentration, total protein level in blood and liver as well as serum and liver content of nitrite anions. LOLA enhances activity of catalase, glutathione S-transferase, manganese superoxide dismutase, increases reduced glutathione level and total antioxidant capacity and decreases thiobarbituric acid reactive substances level. The concomitant use of L-NAME inhibits the action of LOLA to enhance nitrite anions synthesis both in serum and liver, to delay the recovery of hepatocytes, to counteract LOLA effect against blood total protein reduction, to prevent the decline in aminotransferases, alkaline phosphatase,, gamma-glutamyl transferase and glutathione S-transferase activity and to reduce catalase activity and reduced glutathione level. Therefore, in CCl4-induced hepatitis, LOLA effectively prevents cytolysis and cholestasis, improves liver metabolism and protects against oxidative stress. Partially, these changes occur in nitric oxide-mediated mechanism since the use of L-NAME declines most of LOLA effects

Nitric oxide-mediated effects of L-ornithine-L-aspartate in acute toxic liver injury

This study was aimed to investigate nitric oxide-dependent mechanisms of L-ornithine-L-aspartate (LOLA) action in acute toxic liver injury in rats. Acute hepatitis was induced in Wistar rats using 50% oil solution of tetrachloromethane (CCl4) intragastrically (2 g/kg) twice in a 24 hour interval. Intraperitoneal treatment with LOLA (200 mg/kg) was started 6 hours after the second CCl4 administration and maintained for 3 consecutive days. L-Nω-Nitroarginine Methyl Ester (L-NAME) was used intraperitoneally (10 mg/kg). In CCl4-induced hepatitis, LOLA restores the structure of hepatocytes and prevents aminotransferases, alkaline phosphatase and gamma-glutamyl transferase elevation. It decreases total bilirubin concentration but does not affect increased cholesterol level. LOLA augments urea concentration, total protein level in blood and liver as well as serum and liver content of nitrite anions. LOLA enhances activity of catalase, glutathione S-transferase, manganese superoxide dismutase, increases reduced glutathione level and total antioxidant capacity and decreases thiobarbituric acid reactive substances level. The concomitant use of L-NAME inhibits the action of LOLA to enhance nitrite anions synthesis both in serum and liver, to delay the recovery of hepatocytes, to counteract LOLA effect against blood total protein reduction, to prevent the decline in aminotransferases, alkaline phosphatase,, gamma-glutamyl transferase and glutathione S-transferase activity and to reduce catalase activity and reduced glutathione level. Therefore, in CCl4-induced hepatitis, LOLA effectively prevents cytolysis and cholestasis, improves liver metabolism and protects against oxidative stress. Partially, these changes occur in nitric oxide-mediated mechanism since the use of L-NAME declines most of LOLA effects