Simultaneous Determination of 11 Marker Compounds in Gumiganghwal-tang by HPLC-DAD and LC-MS

Gumiganghwal-tang has been used for the treatment of common cold for a long-time. We developed an accurate and sensitive high performance liquid chromatography-diode array detection (HPLC-DAD) and electrospray ionization mass spectrometry method for the simultaneous determination of ferulic acid, baicalin, bergapten, methyl eugenol, glycyrrhizin, oxypeucedanin, wogonin, nodakenin, atractylenolide III, imperatorin, and atractylenolide I in Gumiganghwal-tang samples. The analytes were separated on a Shiseido C18 column (5 µm, 4.6 mm I.D. × 250 mm) with gradient elution with acetonitrile and 0.1% trifluoroacetic acid. Eleven compounds were quantitatively determined by HPLC-DAD and identified by LC-MS data. We also validated this method. The calibration curves of all the compounds showed good linear regression. The limits of detection and the limits of quantification ranged from 0.04 to 0.63 and from 0.12 to 1.92 µg/mL, respectively. The relative standard deviation values of intra- and inter-days of this method represented less than 2.9%. The recoveries were found to be in the range of 90.06 – 107.66%. The developed method has been successfully applied to the analysis of Gumiganghwal-tang samples. The established HPLC method could be used to quality control of Gumiganghwal-tang.

Rapid Isolation of Cyanidin 3-Glucoside and Peonidin 3-Glucoside from Black Rice (Oryza sativa) Using High-Performance Countercurrent Chromatography and Reversed-Phase Column Chromatography

Anthocyanins are water soluble plant pigments which are responsible for the blue, red, pink, violet colors in several plant organs such as flowers, fruits, leaves and roots. In recent years, anthocyanin-rich foods have been favored as dietary supplements and health care products due to diverse biological activities of anthocyanins including antioxidant, anti-allergic, anti-diabetic, anti-microbial, anti-cancer and preventing cardiovascular disease. High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase medium pressure liquid chromatography (RP MPLC) method was applied for the rapid and efficient isolation of cyanidin 3-glucoside (C3G) and peonidin 3-glucoside (P3G) from black rice (Oryza sativa L., Poaceae). The crude black rice extract (500 mg) was subjected to HPCCC using two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/0.01% trifluoroacetic acid (TBME/B/A/0.01% TFA, 1 : 3 : 1 : 5, v/v, flow rate - 4.5 mL/min, reversed phase mode) to give enriched anthocyanin extract (37.4 mg), and enriched anthocyanin extract was sequentially chromatographed on RP-MPLC to yield C3G (16.5 mg) and P3G (8.7 mg). The recovery rate and purity of isolated C3G were 76.0% and 98.2%, respectively, and those of P3G were 58.3% and 96.3%, respectively. The present study indicates that HPCCC coupled with RP-MPLC method is more rapid and efficient than multi-step conventional column chromatography for the separation of anthocyanins.

Phytochemical Analysis of the Phenolic Fat-Suppressing Substances in the Leaves of Lactuca raddeana in 3T3-L1 Adipocytes

Lactuca raddeana (Compositae) is used to treat obesity and complications due to diabetes. The five phenolic compounds including chlorogenic acid, chicoric acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, luteolin were qualitatively identified by LC-ESI-MS analysis. The contents were quantitatively determined by HPLC, under the condition of a Capcell Pak C18 column (5 microm, 250 mm x 4.6 mm i.d.) and a gradient elution of 0.05% trifluoroacetic acid (TFA) and 0.05% TFA in MeOH-H2O (60 : 40). The contents of chicoric acid (100.99 mg/g extract) and luteolin 7-O-glucoside (101. 69 mg/g extract) were high, while those of other three phenolic substances were very low. The 3T3-L1 adipocyte cells treated with chicoric acid and luteolin 7-O-glucuronide significantly suppressed the accumulation of fat, suggesting they are effective against obesity. Since high level of peroxynitrite (ONOO) causes cardiovascular disease in obese patients, its scavenging activity was also studied.

Qualitative and quantitative analysis of fluoxetine hydrochloride by 19F NMR / 药学学报

The chemical shift of fluoxetine hydrochloride appears at delta 14.15 in 19F NMR analysis. The delta moved upfield slightly from 14.158 to 14.145 when the concentration of solution became diluted from 2.00 to 0.05 mmol x L(-1). Spiking test was suggested to confirm the existence of the compound for qualitative analysis. 19F NMR detection sensitivity test illustrated that a concentration of 17 mg in 1 L water could be detected while the sample was scanned 500 times with optimum parameters. In quantitative analysis, standard curve of concentration versus fluorine signal intensity was proposed to determine the amount of fluoxetine. Long capillary tube containing trifluoroacetic acid was used as internal standard for the integration measurements and straight line was obtained with good fitting. Direct additions of trifluoroethanol to fluoxetine solutions gave a poorer standard curve.

A Case of Chemical Burn Caused by Trifluoroacetic Anhydride that Mimicked a Hydrofluoric Acid Burn

A 22-year-old woman was referred to our emergency department for the treatment of a chemical injury on her arm. She had accidentally spilled 99% trifluoroacetic anhydride (TFAA) over her left forearm during an organic chemistry experiment. She visited a primary care unit, and then she was referred to our hospital for inactivation of the released fluoride ions. Her skin lesions were different from those caused by hydrofluoric acid (HF) injury. The injured area showed painful whitish maculae and patchy areas with accentuated rim. No vesiculation and bulla formation was detected. We intradermally injected a 5% solution of calcium through a 24-gauge needle into the burned skin. After the injection, she complained of more severe pain. Although TFAA contains fluorine, it does not release free fluoride ions on contact with the skin, unlike HF. In fact, application of calcium gluconate for TFAA burns is not recommended. Rather, it should be avoided since it increases pain and local abscess formation.

Application of reversed-phase ion-pair chromatography for universal estimation of octanol-water partition coefficients of acid, basic and amphoteric drugs / 药学学报

This paper is to establish a reversed-phase ion-pair chromatography (RP-IPC) method for universal estimation of the octanol/water partition coefficients (logP) of a wide range of structurally diverse compounds including acidic, basic, neutral and amphoteric species. The retention factors corresponding to 100% water (logk(w)) were derived from the linear part of the logk'/phi relationship, using at least four isocratic logk' values containing different organic compositions. The logk(w) parameters obtained were close to the corresponding logP values obtained with the standard "shake flask" methods. The mean deviation for test drugs is 0.31. RP-IPC with trifluoroacetic acid as non classic ion-pair agents can be applicable to determine the logP values for a variety of drug-like molecules with increased accuracy.

Liver Function and Inhaled Anesthetics

The liver is the major site of endogenous and exogenous drug metabolism. The primary result of drug metabolism is the production of more water-soluble and therefore more easily excreted drug metabolites. Drugs are sometimes biotransformed into more reactive metabolites, which may lead to toxicity. Volatile anesthetics, like most drugs, undergo metabolism in the body and are sometimes associated with toxic reactions. Here, author will discuss the metabolism and hepatic toxicity of inhaled anesthetics. Toxicity and liver injury have been reported after repeated exposure on subsequent occasions to different fluorinated anesthetics. This phenomenon of cross-sensitization has also been reported with the chlorofluorocarbon(CFC) replacement agents, the hydrochlorofluorocarbons(HCFCs). Halothane, enflurane, sevoflurane, isoflurane, desflurane are all metabolized to trifluoroacetic acid, which have been reported to induce liver injury in susceptible patients. The propensity to produce liver injury appears to parrel metabolism of the parent drug: halothane(20%) >>>> enflurane(2.5%) >> sevoflurane(1%) > isoflurane(0.2%) > desflurane(0.02%). The use of any anesthetic must be based on its benefits and risks, how it may produce toxicity, and in which patients it may be most safely administered. Nonhalogenated inhaled anesthetics (nitrous oxide, xenon) chemically inert and not metabolized in human tissue. The perfect anesthetic agents dose not exist. But ongoing research attempts to uncover emerging toxicities. Xenon is not currently approved for clinical use. Other than the expense associated with its use, it may be the most ideal anesthetic agent. In general, surgical manipulation or disturbance of the surgical site appears to be more important in decreasing hepatic blood flow than current anesthetic agents such as isoflurane, sevoflurane, and desflurane or technique. However, the clinician is challenged to balance new information with current clinical practices and choice the safest, most effective agents for each patient.

Uso do promestrieno e policresuleno no tratamento da adenose e estenose de vagina, após aplicações de 5-fluorouracil e de ácido tricloroacético em paciente com HPV subclínico / The use of promestriene and policresulen in vaginal adenosis and stenosis treatment after applications of 5-fluoroucacil and trichloroacetic acid in subclinical HPV patient

A freqüência das infecções genítais pelo papiloma vírus humano (HPV) vem aumen- tando de forma importante nos últimos anos, em parte devido a maior sensibilidade e especificidade dos exames de detecção do DNA-HPV Com o aumento na sua freqüência, também aumentam os procedimentos terapêuticas, ainda controversos 'e a possibilidade de algumas complicações, como a adenose vaginal não díetilestilbestroi induzida após uso de ácidos ou de laser de CO, Aqui relatamos um caso de uma paciente que apresentava lesões HPV induzidas em colo do útero e vagina. Após tratamento abrasívo destas lesões, principalmente com ácidos, houve o aparecimento de adenose e de estenose vaginal Neste caso, a conduta foi conservadora através de aplicações tópicas de policresuleno, e posteriormente de promestríeno, com o objetivo de acelerar o processo de reepitelização e de cicatrização. Ao final do tratamento, observou-se uma boa evolução das lesões, não havendo então a necessidade de outras intervenções para restaura ção anatõmica e funcional da vagina.

Influence of Bradykinin on Catecholamine Release from the Rat Adrenal Medulla

The present study was undertaken to investigate the effect of bradykinin on secretion of catecholamines (CA) evoked by stimulation of cholinergic receptors and membrane depolarization from the isolated perfused model of the rat adrenal glands, and to elucidate its mechanism of action. Bradykinin (3 X 10 (-8) M) alone produced a weak secretory response of the CA. however, the perfusion with bradykinin (3 X 10 (-8) M) into an adrenal vein of the rat adrenal gland for 90 min enhanced markedly the secretory responses of CA evoked by ACh (5.32 X 10 (-3) M), excess K+ (5.6 X 10 (-2) M, a membrane depolarizer), DMPP (10 (-4) M, a selective neuronal nicotinic agonist) and McN-A-343 (10 (-4) M, a selective M1-muscarinic agonist). Moreover, bradykinin (3 X 10 (-8) M) in to an adrenal vein for 90 min also augmented the CA release evoked by BAY-K-8644, an activator of the dihydropyridine L-type Ca2+ channels. However, in the presence of (N-Methyl-D-Phe7) -bradykinin trifluoroacetate salt (3 X 10 (-8) M), an antagonist of BK2-bradykinin receptor, bradykinin no longer enhanced the CA secretion evoked by Ach and high potassium whereas the pretreatment with Lys- (des-Arg9, Leu8) -bradykinin trifluoroacetate salt (3 X 10 (-8) M), an antagonist of BK1-bradykinin receptor did fail to affect them. Furthermore, the perfusion with bradykinin (3 X 10 (-6) M) into an adrenal vein of the rabbit adrenal gland for 90 min enhanced markedly the secretory responses of CA evoked by excess K+ (5.6 X 10 (-2) M). Collectively, these experimental results suggest that bradykinin enhances the CA secretion from the rat adrenal medulla evoked by cholinergic stimulation (both nicotininc and muscarinic receptors) and membrane depolarization through the activation of B2-bradykinin receptors, not through B1-bradykinin receptors. This facilitatory effect of bradykinin seems to be associated to the increased Ca2+ influx through the activation of the dihydropyridine L-type Ca2+ channels.

The Effect of Trifluoroacetic Acid, a Metabolite of Isoflurane on the ATP-sensitive Potassium Channel in Rabbit Ventricular Myocytes / 대한마취과학회지

BACKGROUND: Activation of ATP-sensitive K+ channels (KATP channels) in the cardiac muscle produces cardioprotective effects during myocardial ischemia. Previous experimental evidence indicates that volatile anesthetics exert beneficial actions in ischemic myocardium and enhance functional recovery of stunned myocardium. More recently, volatile anesthetics have been demonstrated to produce cardioprotective effects in stunned myocardium in vivo, and these effects are blocked by a KATP channel antagonist. This finding suggests that KATP channel activation by isoflurane may mediate antiischemic effects. However, it was demonstrated that isoflurane inhibited KATP channel activity in rabbit ventricular myocytes. To explain the discrepancy, the present investigation tested the hypothesis that isoflurane and its metabolite, trifluoroacetic acid, contributes to the activation of KATP channels in rabbit ventricular myocytes. METHODS: Single ventricular myocytes were isolated from rabbit hearts by an enzymatic dissociation procedure. Single-channel currents were measured in inside-out patch configurations of the patch-clamp technique. The perfusing liquid was equilibrated with isoflurane by passing 100% O2 through a vaporizer. RESULTS: Isoflurane inhibited KATP channel activity without a change in the single-channel conductivity. Isoflurane decreased the burst duration and increased the interburst duration. In addition, isoflurane diminished the ATP sensitivity of KATP channels. Trifluoroacetic acid, a metabolite of isoflurane, enhanced the channel activity in a dose-dependent fashion. Trifluoroacetic acid increased the burst duration and decreased the interburst duration without a change in the single-channel conductivity. Isoflurane and trifluoroacetic acid diminished the ATP sensitivity of KATP channels. CONCLUSIONS: These results imply that isoflurane and its metabolite could mediate cardioprotective effects via KATP channel activation during myocardial ischemia.

Activation of ATP-sensitive potassium channels by the predominant metabolite of isoflurane in rabbit ventricular myocytes

Background: Recent in vivo experimental evidence suggests that isoflurane-induced cardioprotection may involve KATP channel activation. However, it was demonstrated that isofluran inhibited KATP channel activities in the inside-out patch mode. To explain this discrepancy, the present investigation tested the hypothesis that a metabolite of isoflurane, trifluoroacetic acid (TFA), contributes to isoflurnae-induced cardioprotection via KATP channel activation during myocardial ischemia and reperfusion. Methods: Single ventricular myocytes were isolated from rabbit hearts by an enzymatic dissociation procedure. Patch-clamp techniques were used to record single-channel currents. KATP channel activities were assessed before and after the application of TFA with the inside-out patch mode. Results: TFA enhanced channel activity in a concentration-dependent fashion. The concentration of TFA for half-maximal activation and the Hill coefficient were 0.03 mM and 1.2, respectively. TFA did not affect the single channel conductance of KATP channels. Analysis of open and closed time distributions showed that TFA increased burst duration and decreased the interburst interval without changes in open and closed time distributions shorter than 5 ms. TFA diminished ATP sensitivity of KATP channels in a concentration-response relationship for ATP. Conclusions: TFA, a metabolite of isoflurane, enhanced KATP channel activity in a concentration-dependent fashion. These results imply that TFA could mediate isoflurane-induced cardioprotection via KATP channel activation during myocardial ischemia and reperfusion.

Antitumor effects of rhodium (II) complexes on mice bearing Ehrlich tumors

Rhodium (II) trifluoracetate (TFARh), rhodium (II) trifluoracetate adduct with sulfadiazine (TFARh.Sd) and rhodium (II) acetate adduct with sulfisoxazole (RhSx) were tested in mice for acute toxicity, antitumoral activity against Ehrlich ascites carcinoma and for viability of Ehrlich tumor cells in culture. At ip doses up to 60 µmg/kg (40-70 and 59 mg/kg, respectively), these coumpounds had no toxic effects up to 14 days. At ip doses of 10 µmol Kg-1 day-1 for 5 days, TFARh and TFARh.Sd significantly increased the survival rate of mice bearing Ehrlich ascites cells (probability of survival to the end of 34th day, controls = 0.23, TFARh = 0.85, TFARh.Sd = 0.74). No significant effect was observed for RhSx. In vitro, these rhodium complexes at 40 µM significantly increased the number of dead cells in cultured Ehrlich tumor cells

Exposiçäo profissional aos anestésicos voláteis: 2. monitorizaçäo ambiental e monitorizaçäo biológica / Occupational exposure as volatile anesthetics: 2. physiologic and environmental monitoring

Säo apresentados vários aspectos da monitorizaçäo ambiental e monitorizaçao biológica, valores-limites propostos e resultados laboratoriais de amostras coletadas durante a realizaçäo de cirurgias.

Efeito de alguns fatores físicos sobre a estabilidade de vitamina A e carotenóides plasmáticos / Effect of some physical factors on the stability of vitamin A and plasma carotenoids

Efeito de alguns fatores físicos sobre a estabilidade de vitamina A e carotenóides plasmáticos. Foi testada a estabilidade da vitamina A e dos carotenóides em plasma, em diferentes condiçöes de temperatura, luz e evaporaçäo, além de verificar os efeitos de congelamentos-descongelamentos cíclicos. Para as dosagens de vitamina A foi utilizado o macro-método de Neeld-Pearson e os ensaios foram realizados em triplicata. Os resultados sugerem que houve alteraçäo apenas após exposiçäo solar à luz direta, fator que deve ser considerado na realizaçäo de trabalhos de campo