Development and validation of a simultaneous analytical method for non-steroidal therapeutic compounds in cosmetics using liquid chromatography-tandem mass spectrometry.

Atopic dermatitis is a typical chronic inflammatory skin disease that affects all age groups and requires basic skin care for treatment. Anti-inflammatory and antiallergy steroids are the most frequently used treatments but they are limited due to their side effects caused by a weakening of the immune system. Many consumers focus on performance as a criterion for selecting cosmetics. However, steroids have been illegally used to improve the performance of cosmetics, and consumers have been adversely affected by the corresponding side effects. In this paper, we propose a simple and rapid method using liquid chromatography-tandem mass spectrometry to simultaneously analyze ten non-permitted atopic therapeutic compounds in cosmetic products: chlorpheniramine maleate, ketotifen fumarate, doxepin hydrochloride, azelastine hydrochloride, bufexamac, clotrimazole, tranilast, fusidic acid, tacrolimus, and pimecrolimus. Additionally, the major characteristic fragment ions for tacrolimus, pimecrolimus, and clotrimazole were identified by time-of-flight mass spectrometry. The specificity, linearity, limit of detection, limit of quantification, recovery, precision, accuracy, and stability of the proposed method were validated. The limit of detection and quantification were in the ranges of 5.05-203.30 pg/mL and 15.15-609.90 pg/mL, respectively. The proposed analysis method could help improve the safety management of cosmetics.

Characterization and antioxidant activity of avenanthramides from selected oat lines developed by mutagenesis technique.

From a mutagenized oat population, produced by ethyl methanesulfonate mutagenesis, hulled grains from 17 lines with elevated avenanthramide (AVN) content were selected and their AVN structures, concentrations and antioxidant potentials were determined by HPLC-MS2 and HPLC equipped with an on-line ABTS+ antioxidant detection system. The data obtained showed qualitative and quantitative differences in the synthesis of AVNs in the different lines, with a total AVN concentration up to 227.5 µg/g oat seed flour in the highest line, compared with 78.2 µg/g seed in the commercial line, SW Belinda. In total, 25 different AVNs were identified with avenanthramide B structures being among the most abundant, and AVN C structures having the highest antioxidant activity. The findings indicate the potential of oat mutagenesis in combination with a high precision biochemical selection method for the generation of stable mutagenized lines with a high concentration of total and/or individual AVNs in the oat seed grain.

Comprehensive analysis of oat avenanthramides using hybrid quadrupole-Orbitrap mass spectrometry: Possible detection of new compounds.

RATIONALE: Avenanthramides (AVNs) are constituents unique to oats and have many outstanding health benefits. AVNs are antioxidants and possess anti-inflammatory, antifungal and antibacterial activity. The number of known AVNs increased recently because of the latest developments in high-resolution tandem mass spectrometry (HRMS/MS) techniques. METHODS: Oat seed extract from 10 oat cultivars was analysed using ultra-high-performance liquid chromatography (UHPLC) and Q Exactive hybrid quadrupole-Orbitrap mass spectrometry (HRMS/MS) with positive heated electrospray ionization. RESULTS: Thirty-five AVNs were identified and characterized in seed extracts, and the structures of 10 novel AVNs were tentatively elucidated, among which were AVNs bearing a cinamoyl or sinapoyl moiety. These AVNs are reported in oats for the first time. The method was validated using AVN standards (AVNs 2c, 2f and 2p), with limits of detection and quantitation at low picomole levels. Recovery of AVN standards varied from 83% to 106%, and relative standard deviations ranged from 2% to 9%. The total AVNs in the selected oat varieties ranged from 36.0 to 302.5 µg/g (dry weight), with AVN 2c, AVN 2f and AVN 2p representing approximately 65%-70% of that total. CONCLUSIONS: Our comprehensive method for detecting the full avenanthramide spectrum can contribute to better understanding the chemical and biological properties of individual AVNs for utilization in developing new oat cultivars and novel functional foods.

A simple method to discriminate Guangchenpi and Chenpi by high-performance thin-layer chromatography and high-performance liquid chromatography based on analysis of dimethyl anthranilate.

Citri Reticulatae Pericarpium (CRP), the dried pericarp of Citrus reticulata Blanco, can be divided into "Guangchenpi" (GCP, the dried pericarps derived from Citrus reticulata 'Chachi') and "Chenpi" (CP, the dried pericarps derived from other cultivars of Citrus reticulata Blanco). To discriminate between GCP and CP, a simple and reliable high-performance thin-layer chromatography (HPTLC) method was firstly developed to analyze the volatile compound dimethyl anthranilate, and a high-performance liquid chromatography (HPLC) method was established to simultaneously quantify dimethyl anthranilate and three predominant flavonoids (hesperidin, nobiletin and tangeretin) in CRP samples. Both the HPTLC analysis and HPLC-orthogonal partial least squares discrimination analysis (OPLS-DA) indicated that GCP can be effectively distinguished from CP based on analysis of dimethyl anthranilate. Our results indicated that dimethyl anthranilate can be used as a marker compound for discrimination of GCP and CP. This work provided a convenient approach which might be applied for quality evaluation of CRP.

Oat germination and ultrafiltration process improves the polyphenol and avenanthramide contents with protective effect in oxidative-damaged HepG2 cells.

Oat is the nutritious crop containing various compounds with antioxidant properties, such as polyphenols. In this study, we investigated the effect of germination and ultrafiltration process on polyphenol and avenanthramide contents in oat as well as their cytoprotective effect. Germination of oat for 48 hr significantly increased avenanthramide (5.5 to 11.3 mg/g) and polyphenol (115 to 155 mg GAE/g) contents. The compounds were more concentrated after ultrafiltration using 10 kDa membranes (polyphenol, 206 GAE/g; avenanthramide, 18 mg/g). In addition, oat extracts significantly reduced the cellular ROS level against tert-butyl hydroperoxide (t-BHP) stimulation in HepG2 cells. In the mechanistic study, oat extracts induced Nrf2 translocation to the nucleus by inhibition of Keap1 expression, resulting into upregulation of γ-GCS and NQO1. In conclusion, oat germination and ultrafiltration processes increased the polyphenol content, including that of avenanthramide. These extracts protected cells from t-BHP by radical scavenging activities and induced Nrf2 pathway activation. PRACTICAL APPLICATIONS: This study presents the method for avenanthramide-concentrated extract which is unique bioactive compounds in oat. In addition, antioxidant activity and their mechanisms of the avenanthramide-enriched extracts were evaluated. The polyphenol compounds including avenanthramide were found to increase after germination and ultrafiltration, thereby improving the radical scavenging ability. These results can be utilized as data for the development of health-promoting materials using oats.

Mass spectrometric characterisation of avenanthramides and enhancing their production by germination of oat (Avena sativa).

Avenanthramides are amides, with a phenylalkenoic acid (PA) and an anthranilic acid (AA) subunit, which are secondary metabolites of oat. Oat seeds were germinated, extracted, and the avenanthramides analysed by a combination of UHPLC with ion trap and high resolution ESI-MS. Typical fragmentation pathways with corresponding diagnostic fragments belonging to the PA and AA subunits were identified and summarised in a decision guideline. Based on these findings 28 unique avenanthramides were annotated in the oat seed(ling) extracts, including the new avenanthramide 6f (with a 4/5-methoxy AA subunit). Avenanthramide content increased by 25 times from seed to seedling. Avenanthramides 2p, 2c, and 2f, which are commonly described as the major avenanthramides, represented less than 20% of the total content in the seedlings. Future quantitative analyses should, therefore, include a wider range of avenanthramides to avoid underestimation of the total avenanthramide content.

High Levels of Avenanthramides in Oat-Based Diet Further Suppress High Fat Diet-Induced Atherosclerosis in Ldlr-/- Mice.

Oats, in addition to cholesterol-lowering properties, contain unique antioxidants called avenanthramides (Avns), which inhibit both inflammatory cytokines and adhesion molecules in endothelial cells in culture. This study evaluated the effects of Avns of oats on atherosclerosis in Ldlr-/- mice, one of the most commonly used atherosclerosis mouse models with their similar cholesterol distributions to humans. The Ldlr-/- mice were fed a low fat, high fat, high fat containing regular oat brans with low levels of Avns (HFLA), or high fat containing regular oat brans with high levels of Avns (HFHA) diet. After 16 weeks of intervention, blood cholesterol and extent of aortic lesions were evaluated. We found that both oat-based diets reduced high fat diet-induced atheroma lesions in the aortic valve (p < 0.01). Furthermore, the effects of oat-based diets are more profound in HFHA mice than mice fed HFLA. Total plasma cholesterol levels were similarly reduced in both oat-supplemented mice. We concluded that oat bran diets reduce atheroma lesions and higher levels of Avns further reduce aortic lesions compared to regular oat bran. These preliminary in vivo data indicate that consumption of oats bran, with high Avns, has demonstrable beneficial effects on prevention of cardiovascular disease.

Dissipation and residues of the diamide insecticide chlorantraniliprole in ginseng ecosystems under different cultivation environments.

Ginseng is a valuable herb, the roots of which have been consumed for medicinal use since ancient times in Asian countries. Currently, its cultivation in China is conducted in either forests or non-forest areas. In this study, we investigated the dissipation and residues of the diamide insecticide chlorantraniliprole (CAP) in ginseng ecosystems in these two cultivation environments. The half-life of CAP in ginseng leaves was calculated to be 16.5 days in non-forest areas and 23.9 days in forests, while the half-life of CAP in the soil under non-forest and forest cultivation was 33.0 and 53.3 days, respectively. The relatively long persistence of CAP in ginseng leaves and soil could be attributed to the unique cultivation techniques of ginseng, which prevents exposure to direct sunlight irradiation and rainfall. The residual amounts of CAP in ginseng leaves, roots, and soil in non-forest areas were 0.168, 0.011, and 0.013 mg kg-1 21 days after CAP application at the maximum label-allowed dosage and frequency versus 0.564, 0.013, and < 0.005 mg kg-1 in forests. CAP application strategies for the control of Lepidoptera pests such as armyworms, and the safety of harvested ginseng roots are recommended for ginseng cultivated in forests and non-forest areas based on the results of the CAP residue experiments.

Chromatographic Methods to Evaluate Nutritional Quality in Oat.

Oats (A. sativa L.) have an important and positive role in human diet and health. The health benefits of oats are attributed to its multifunctional characteristic and nutritional profile, being an important source of soluble dietary fiber, well-balanced proteins, unsaturated fatty acids, vitamins, essential minerals, and a good source of natural antioxidants. These antioxidants include the avenanthramides (Avns) and avenalumic acids, which are unique to oats among cereals. High-performance liquid chromatography allows a simultaneous quantification of free amino acids and biogenic amines in oat samples as their OPA/FMOC-CL (o-phthalaldehyde/9-fluorenylmethoxycarbonyl chloride) derivatives. In addition, an ultra-performance liquid chromatography/mass spectrometry method was developed to quantify and characterize avenanthramides contained in oat samples.

Validation of chemical compound library screening for transcriptional co-activator with PDZ-binding motif inhibitors using GFP-fused transcriptional co-activator with PDZ-binding motif.

Transcriptional co-activator with PDZ-binding motif (TAZ) plays versatile roles in cell proliferation and differentiation. It is phosphorylated by large tumor suppressor kinases, the core kinases of the tumor-suppressive Hippo pathway. Phosphorylation induces the cytoplasmic accumulation of TAZ and its degradation. In human cancers, the deregulation of the Hippo pathway and gene amplification enhance TAZ activity. TAZ interacts with TEA domain family members (TEAD), and upregulates genes implicated in epithelial-mesenchymal transition. It also confers stemness to cancer cells. Thus, TAZ activation provides cancer cells with malignant properties and worsens the clinical prognosis. Therefore, TAZ attracts attention as a therapeutic target in cancer therapy. We applied 18 606 small chemical compounds to human osteosarcoma U2OS cells expressing GFP-fused TAZ (GFP-TAZ), monitored the subcellular localization of GFP-TAZ, and selected 33 compounds that shifted GFP-TAZ to the cytoplasm. Unexpectedly, only a limited number of compounds suppressed TAZ-mediated enhancement of TEAD-responsive reporter activity. Moreover, the compounds that weakened TEAD reporter activity did not necessarily decrease the unphosphorylated TAZ. In this study, we focused on three compounds that decreased both TEAD reporter activity and unphosphorylated TAZ, and treated several human cancer cells with these compounds. One compound did not show a remarkable effect, whereas the other two compounds compromised the cell viability in certain cancer cells. In conclusion, the GFP-TAZ-based assay can be used as the first screening for compounds that inhibit TAZ and show anticancer properties. To develop anticancer drugs, we need additional assays to select the compounds.

Identification of a new antinociceptive alkaloid isopropyl N-methylanthranilate from the essential oil of Choisya ternata Kunth.

ETHNOPHARMACOLOGICAL RELEVANCE: Mexican people employed infusion of leaves of Choisya ternata Kunth for their antispasmodic and "simulative properties". AIM OF THE STUDY: In the present study the detailed GC and GC-MS analyses of the essential oil of Choisya ternata Kunth (Rutaceae) were performed. The presence of a minor constituent isopropyl N-methylanthranilate (1) was revealed among other identified volatiles. A synthesis of 1 was undertaken in order to corroborate this find and obtain gram quantities that would allow the testing of its biological activity (peripheral and central antinociceptive activity). MATERIALS AND METHODS: The oils were investigated by GC and GC-MS. Synthesized compounds were spectrally characterized (UV-Vis, IR, 1D and 2D NMR, MS). The obtained synthetic samples of compounds were assayed for peripheral and central antinociceptive activity in two models (effects on acetic acid induced writhing in mice and the hot plate test for nociception). RESULTS: Detailed GC and GC-MS analyses of the essential oil of Choisya ternata Kunth (Rutaceae) among 157 other identified volatiles revealed the presence of a minor constituent isopropyl N-methylanthranilate (1). Compound 1, named ternanthranin, is therefore detected as a natural product for the first time with a very restricted occurrence (samples of several citrus oils were screened for the presence of 1). The antinociceptive activities were assayed for ternanthranin, the two other synthetic analogs, methyl and propyl N-methylanthranilate, as well as the essential oil and the crude ethanol extract of the leaves. The results clearly demonstrate a very high (even significant at 0.3 mg/kg) dose dependent activity for the anthranilates (and the extracts). Isopropyl N-methylanthranilate showed the highest, while methyl N-methylanthranilate showed the lowest activity (with the methyl ester at 3 mg/kg still better than acetylsalicylic acid, at 200 mg/kg, in the first, or comparable with morphine, at 5mg/kg, in the second test). CONCLUSION: This study once again revealed that detailed investigations of plant species with ethnopharmacologically documented activity may yield new natural compounds-a new alkaloid (ternanthranin), a volatile simple anthranilate that can be considered responsible for the antinociceptive activity of the crude plant extracts.

The green oat story: possible mechanisms of green color formation in oat products during cooking.

Consumers occasionally report greenish colors generated in their oat products when cooking in tap water. Here we have investigated pH and ferrous (Fe(2+)) ion as possible mechanisms for this color change. Steel-cut oat groats can turn brown-green color when cooked in alkaline conditions (pHs 9 to 12). Extraction of this color with methanol, and high-pressure liquid chromatography indicated a direct association of this color with the phenolic acid or avenanthramide content of the oat. The presence of 50 mM NaHCO(3) in water will cause oat/water mixtures to turn alkaline when cooked as CO(2) is driven off, generating OH(-) ion. Although tap water rarely, if ever, contains so much bicarbonate, bicarbonate is used as a leavening agent in baking applications. Industrial interests using baking soda or alkaline conditions during oat processing should be aware of possible off color generation. We have also found that as little as 10 ppm Fe(2+) will turn oat products gray-green when cooked. The aleurone stained darker than the starchy endosperm. Other divalent cations, such as Ca(2+) or Mg(2+) had no effect on cooked oat color. As much as 50 ppm Fe(2+) may be found in freshly pumped well water, but Fe(2+) reacts quickly with oxygen and precipitates as Fe(OH)(3). Thus, some freshly pumped well water may turn oats green when cooked, but if the water is left under atmospheric conditions for several hours, no discoloration will appear in the cooked oats.

[Determination of salylic acid, syringic acid, benzoic acid and anthranilic acid in Radix Isatidis by HPCE].

OBJECTIVE: To develop a simple and rapid capillary electrophoresis (CE) method for the separation and determination of four active organic acids including salicylic acid, syringic acid, benzoic acid, and anthranilic acid in Radix Isatidis. METHOD: The HPCE system consisted of a fused-silica capillary column of 47.3 cm (38.3 cm to the detector) x50 microm i.d. and a mixture ofacetonitrile-borate buffer (15% acetonitrile, 25 mmol L(-1) borate, 15 mmol L(-1) beta-CD, pH 9.10) solution as the operating buffer. The applied voltage was 11.5 kV and the UV detection was set at 220 nm. The effects of the applied voltage, detection wavelength, and the pH of buffer, the concentration of buffer, acetonitrile and beta-CD were investigated. RESULT: The linear calibration rang was 3.0-90 mg L(-1) (r=0.9994) for salylic acid, 4.0-120 mg L(-1) (r=0.9995) for syringic acid, 2.0-60 mg L(-1) (r=0.9998) for benzoic acid and 5.0-100 mg L(-1) (r=0.9992) for anthranilic acid. The recoveries of salylic acid, syringic acid, benzoic acid and anthranilic acid were 95.9%-102.6%, 98.6%-103.4%, 98.7%-104.1%, 96.1%-104.3% respectively. The detection limits of salylic acid, syringic acid, benzoic acid and anthranilic acid were 0.7, 1.1, 1.2 and 1.5 mg L(-1), respectively.

Production and analysis of organic acids in hairy-root cultures of Isatis indigotica Fort. (indigo woad).

Hairy roots were induced from both cotyledon and hypocotyl explants of Isatis indigotica Fort. (indigo woad) through transformation with Agrobaterium rhizogenes strain A4, R1601 and ATCC15834. The results showed that the cotyledons were the preferred explants to hypocotyls and A4 was the most suitable A. rhizogenes strain for the transformation and induction of hairy roots of I. indigotica. High-voltage paper electrophoresis (HVPE) analysis demonstrated the production of mannopine in hairy roots and confirmed the successful transfer of Ri T-DNA (root-inducing transferred DNA) of A. rhizogenes into the I. indigotica genome. Five organic acids, namely CPQ [3-(2-carboxyphenol)-4(3 H )-quinazolinone], syringic acid, salicylic acid, benzoic acid and 2-aminobenzoic acid, which were considered as main antiviral components of I. indigotica, were detected in natural roots, hairy roots and liquid media with high-performance capillary electrophoresis. The results showed CPQ production in hairy roots was significantly higher than that in natural roots. Our results also revealed that all the five organic acids could be excreted from hairy roots into liquid media, and the concentrations of organic acids in the liquid media paralleled those in hairy roots. The hairy roots of I. indigotica grew fast and showed an S-shaped growth curve that reached its apex on the day 24 of culture with a 20-fold increase in fresh weight compared with the starting inoculums. The accumulation of the two organic acids CPQ and syringic acid in liquid media paralleled the growth of hairy roots. MS [Murashige, T. and Skoog, F. (1962) Physiol. Plant. 15, 473-497] medium or half-strength MS medium supplemented with 30 g/l maltose was found to be best for hairy-root culture and accumulation of CPQ.

Leaf volatile compounds of seven citrus somatic tetraploid hybrids sharing willow leaf mandarin (Citrus deliciosa Ten.) as their common parent.

Volatile compounds were extracted by a pentane/ether (1:1) mixture from the leaves of seven citrus somatic tetraploid hybrids sharing mandarin as their common parent and having lime, Eurêka lemon, lac lemon, sweet orange, grapefruit, kumquat, or poncirus as the other parent. Extracts were examined by GC-MS and compared with those of their respective parents. All hybrids were like their mandarin parent, and unlike their nonmandarin parents, in being unable to synthesize monoterpene aldehydes and alcohols. The hybrids did retain the ability, although strongly reduced, of their nonmandarin parents to synthesize sesquiterpene hydrocarbons, alcohols, and aldehydes. These results suggest that complex forms of dominance in the mandarin genome determine the biosynthesis pathways of volatile compounds in tetraploid hybrids. A down-regulation of the biosynthesis of methyl N-methylanthranilate, a mandarin-specific compound, originates from the genomes of the nonmandarin parents. Statistical analyses showed that all of the hybrids were similar to their common mandarin parent in the relative composition of their volatile compounds.

Avenanthramides in oats (Avena sativa L.) and structure-antioxidant activity relationships.

Eight avenanthramides, amides of anthranilic acid (1) and 5-hydroxyanthranilic acid (2), respectively, and the four cinnamic acids p-coumaric (p), caffeic (c), ferulic (f), and sinapic (s) acid, were synthesized for identification in oat extracts and for structure-antioxidant activity studies. Three compounds (2p, 2c, and 2f) were found in oat extracts. As assessed by the reactivity toward 1,1-diphenyl-2-picrylhydrazyl (DPPH), all avenanthramides except 1p showed activity. Initially, the antioxidant activity of the avenanthramides decreased in a similar order as for the corresponding cinnamic acids, that is: sinapic > caffeic > ferulic > p-coumaric acid. The avenanthramides derived from 2 were usually slightly more active than those derived from 1. All avenanthramides inhibited azo-initiated peroxidation of linoleic acid. 1c and 1s were initially the most effective compounds. The relative order of antioxidant activities was slightly different for the DPPH and the linoleic acid assays run in methanol and chlorobenzene, respectively.

Identification of potent odorants in Chinese jasmine green tea scented with flowers of Jasminum sambac.

The odorants in Chinese jasmine green tea scented with jasmine flowers (Jasminum sambac) were separated from the infusion by adsorption to Porapak Q resin. Among the 66 compounds identified by GC and GC/MS, linalool (floral), methyl anthranilate (grape-like), 4-hexanolide (sweet), 4-nonanolide (sweet), (E)-2-hexenyl hexanoate (green), and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (sweet) were extracted as potent odorants by an aroma extract dilution analysis and sensory analysis. The enantiomeric ratios of linalool in jasmine tea and Jasminum sambac were determined by a chiral analysis for the first time in this study: 81.6% ee and 100% ee for the (R)-(-)-configuration, respectively. The jasmine tea flavor could be closely duplicated by a model mixture containing these six compounds on the basis of a sensory analysis. The omission of methyl anthranilate and the replacement of (R)-(-)-linalool by (S)-(+)-linalool led to great changes in the odor of the model. These two compounds were determined to be the key odorants of the jasmine tea flavor.

[Flavonoid and dianthranoid levels of the St.-John's-wort flowering tops]. / Teneurs en flavonoïdes et en dianthranoïdes de la sommité fleurie de millepertuis.

Dried flowering tops of Hypericum perforatum L. (16 batches) and H. maculatum (3 batches) were studied according to the harvest period. The mean levels of the flavonoid and dianthranoid compounds were respectively: total flavonoids 3.20 and 3.92%, hyperoside 0.87 and 2. 12%, rutin 0.54 and 0.02%, isoquercitrin 0.48 and 0.75%; total dianthranoids 0.14 and 0.13%, hypericin 0.04 and 0.02%, pseudohypericin 0.08 and 0.11%. The flavonoid and dianthranoid levels were higher in young flowering tops of H. perforatum. 3 commercial batches were also examined for a comparative study. Pharmacopoeial specifications are proposed for a revision of the monograph "St.-John's-wort".

Occurrence of avenanthramides and hydroxycinnamoyl-CoA:hydroxyanthranilate N-hydroxycinnamoyltransferase activity in oat seeds.

Oat phytoalexins, avenanthramides, occur as constitutive components in seeds. The amounts of each avenanthramide were analyzed. The composition of avenanthramides in dry seeds was different from that in elicitor-treated leaves. In seeds, avenanthramide C was most abundant with an amount two times larger than that of avenanthramide A or B. On the other hand, avenanthramide A was the major component in elicitor-treated leaves. The total amount of avenanthramides in seeds increased 2.5 times during imbibition for 48 h although the composition did not change. The hydroxycinnamoyl-CoA:hydroxyanthranilate N-hydroxycinnamoyltransferase (HHT, EC 2.3.1.-) activity, which is responsible for the final condensation step in the avenanthramide biosynthesis, was detected in dry seeds. The activity was localized in endosperm and scutellum, and slightly increased during 48-h imbibition. The enzyme was partially purified by anion exchange chromatography from both dry seeds and elicitor-treated leaves The activity was separated into two peaks by chromatography, indicating that HHT consists of at least two isoforms. The substrate specificities of HHT isoforms from seeds were different from each other.

Técnica analítica para el control de calidad de las tabletas de Ribofen 80 mg / Analytical technique for the quality control of ribofen tablets 80 mg

Se desarrolló una técnica para el control de calidad de las tabletas de ribofen, con el empleo de la espectrofotometría UV y la cromatografía líquida de alta resolución, que incluye la identificación, disolución, uniformidad de dosis y valoración. Las tabletas cumplen con los índices de calidad establecidos en la técnica. Se estudio la especificidad, linealidad, precisión, exactitud y se corroboró la validez de estos parámetros en ambos métodos