Validation of a Novel RP-HPLC Technique for Simultaneous Estimation of Lignocaine Hydrochloride and Tibezonium Iodide: Greenness Estimation Using AGREE Penalties.

Herein, we reported an HPLC method for the simultaneous determination of tibezonium iodide (TBN) and lignocaine hydrochloride (LGN). The method was developed according to the International Conference for Harmonization guidelines (ICH) Q2R1 using Agilent® 1260 with a mobile phase consisting of acetonitrile and phosphate buffer (pH 4.5) in a volumetric ratio of 70:30 and flowing through a C8 Agilent® column at 1 mL/min. The results revealed that TBN and LGN peaks were isolated at 4.20 and 2.33 min, respectively, with a resolution of 2.59. The accuracy of TBN and LGN was calculated to be 100.01 ± 1.72% and 99.05 ± 0.65% at 100% concentration, respectively. Similarly, the respective precision was 100.03 ± 1.61% and 99.05 ± 0.48%. The repeatability for TBN and LGN was found to be 99.05 ± 0.48% and 99.19 ± 1.72%, respectively, indicating that the method was precise. The respective regression co-efficient (r2) for TBN and LGN was found to be 0.9995 and 0.9992. Moreover, the LOD and LOQ values for TBN were 0.012 and 0.037 µg/mL, respectively, while for LGN, they were 0.115 and 0.384 µg/mL, respectively. The calculated greenness of the method for ecological safety was found to be 0.83, depicting a green contour on the AGREE scale. No interfering peaks were found when the analyte was estimated in dosage form and in volunteers' saliva, depicting the specificity of the method. Conclusively, a robust, fast, accurate, precise and specific method was successfully validated to estimate TBN and LGN.

GC-MS analysis, anticancer and anti-inflammatory activities of Saussurea hypoleuca spreng. Root.

Study has been premeditated to appraise the anticancer and anti-inflammatory activities of a native medicinal plant Saussurea hypoleuca Spreng root. Anticancer assays including MTT, Alamar Blue (AB), Neutral Red (NR) & LDH were employed on root methanolic extract (RME) and all fractions to calculate % age of cell viability and cell cytotoxicity. All fractions of plant root were tested for in vitro as well as in vivo anti-inflammatory assays by reported methods. GC-MS analysis of n-hexane: chloroform fractions in column chromatography has shown isopropyl myristate, hexadecanoic acid, 11-octadecenoic acid, Di-n-octyl phthalate, dioctyl ether, decanedioic acid, 1H-3a,7-Methanoazulene, 3,4-hexanedione and Tetracosapentaene. Percentage of cell viability in anticancer assays was significantly high in all fractions. However, whole results were momentous with ethyl acetate and aqueous fractions owning to excellent profile in evaluating cytotoxicity in each assay. COX-2 inhibition was calculated which was high in RME (68.69%), ethyl acetate (56.52%), aqueous (55.21%) and chloroform fraction (53.47%). Carrageenan and formalin models were developed on rats to investigate in vivo anti-inflammatory activity. RME (56.19%, 71.09%, 66.4%, 67.99%) and ethyl acetate (51.36%, 64.97%, 55.63% & 61.01%) produced significant % age inhibition in dose dependent manner at 200 and 400 mg/kg doses respectively. All above findings direct that plant root holds strong anticancer and anti-inflammatory activities.

Development and validation of single analytical HPLC method for determination of flavoxate HCl in bulk, tablets and biological fluids.

A simple, sensitive and precise high performance liquid chromatographic (HPLC) method was developed and validated for determination of flavoxate HCI in raw material, tablets and biological fluids. The method followed by using the Zorbax XDB-C18 column containing Di-isobutyl n-octadeceylsilane (4.6mm×150mm, 5µm). The mobile phase consisted of acetonitrile: methanol: 0.15M sodium perchlorate (17:35:48 v/v) having pH 3. UV detection was carried out at 229nm at 40°C. Results indicated that the method has successfully established and validated in accordance with ICH guidelines acceptance criteria for linearity (0.03-7.5µg), accuracy (101.18-101.28%), robustness of column age and column lot (peak area %CV<0.04, purity %CV< 0.006) and robustness of HPLC condition (%CV<0.02), precision (intra and inter day precision assay, %CV values for peak area and percent purity of flavoxate HCl<2%) and system suitability parameters. The average noise, theoretical LOD and LOQ were found to be 0.01 mAU, 0.03 mAU and 0.6ng, respectively. The Coefficient of determination (r2) ranging from 0.03µg to 7.5µg, 0.99 which was within acceptable criteria of r2 & gt 0.99. The spiked recoveries of samples were 101.28, 101.18 and 101.18% respectively. All data revealed that this method can be used for in-vitro & in-vivo determination of flavoxate HCI in various pharmaceutical preparations.

Hypoglycemic activity of Ficus racemosa bark in combination with oral hypoglycemic drug in diabetic human.

Medicinal herbs, used in indigenous medicines in crude forms for the management of diabetes mellitus, contain both the organic and inorganic constituents. The aim of the study was to find out the hypoglycemic effect of Ficus racemosa in a group of diabetic subjects taking oral hypoglycemic drug. Twenty five of each, male and female, diabetic patients, selected from Fatima Jinnah Medical College, Lahore, Pakistan, taking oral hypoglycemic drug were included in this study and were given orally the extract (5 mL) of bark of Ficus racemosa (about 100 mg) two times for 15 days. Blood samples for estimation of blood glucose and parameters of liver and renal functions were estimated. It was observed that after taking the herb in combination with drug, blood glucose level (fasting and after breakfast) was markedly decreased in both male and female but significant difference was only observed in sugar level of males after 1.5 h after breakfast. To rule out herb toxicity, liver and renal functions tests of patients was also performed which were observed to be in normal range. Present investigation established a pharmacological evidence to support the folklore claim that Ficus racemosa is good anti-diabetic agent.