Response surface methodology-based optimization of ultrasound-assisted extraction of β-sitosterol and lupeol from astragalus atropilosus (roots) and validation by HPTLC method

Objective: To optimize the ultrasonication method for efficient extraction of P-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology (RSM), and its validation by high performance thin layer chromatography (HPTLC) method. Methods: Ultrasonication method was used to extract β-sitosterol and lupeol from Astragalus atropilosus (roots). RSM was used to optimize the different extraction parameters viz. liquid to solid ratio (10-14 mL/g), temperature (60-80 °C) and time (40-60 min) to maximize the yield of β-sitosterol and lupeol. The quantitative estimation of β-sitosterol and lupeol was done in chloroform extract of Astragalus atropilosus by validated HPTLC method on 10 cm × 20 cm glass-backed silica gel 60F

Quantification of glycyrrhizin in anti-stress herbal formulations by validate HPTLC method: a rational paradigm towards quality control of herbals

In the present study an analytical method of high-performance thin-layer chromatography [HPTLC] has been developed for quantification of glycyrrhizin for marketed antistress liquorice root capsules [LRC] and herbal tea [HT]. Chromatography was performed by using mobile phase ethyl acetate [EA]: glacial acetic acid [GAA]: Methanol [MeOH]: water [H2O] in proportion of 6:2:2:1, v/v/v/v. The developed plate was scanned and quantified densitometrically at absorption maxima 254nm. The method was validated for various analytical parameters viz. precision, accuracy, recovery, robustness, specificity, detection and quantification limits. The developed system was found to give compact spot for glycyrrhizin [Rf = 0.33 +/- 0.001]. The linearity relationship was described by the equation Y=6.841X+ 70.428. The limit of detection [34 ng band-1], limit of quantification [101ng band-1], recovery [99.4-99.8%], and precision [

Quantitative analysis of biomarker rutin in different species of genus Ficus by validated NP and RP-HPTLC methods

Biomarker rutin was analyzed in methanol extracts of leaves of five different species of genus Ficus [Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata and Ficus vasta] by NP-HPTLC [Method I] and RP- HPTLC methods [Method II]. The development and validation for method I was carried out with silica gel 60F[254] plates using EA: GAA: FA: H[2]O [10:1:1:2.5, v/v/v/v] as developing system. Method II was carried out on silica gel 60F[254] RP-18 plates using mobile phase ACN: H[2]O [4:6 v/v]. Both analyses were scanned at 305 nm and were found to give well resolved peak of rutin at Rf 0.28 +/- 0.01 and 0.68 +/- 0.03 for Method I and Method II, respectively. The percentage of rutin was found to be 0.51% and 0.66% in F. ingens, 0.24% and 0.54% in F. palmata and 0.14% and 0.17% in F. vasta by Method I and Method II, respectively. Method II [RP-HPTLC] was found to be more accurate, precise and sensitive than Method I. Method II can be used as an important tool for standardization and quality control of bulk drugs and in-process formulations of rutin

Medico-legal assessment of non-fatal cases of fall from height in Karachi

Background: Fall from height is a tragedy, largely avertable, often leading to death or morbidity. Apart from the physical dimension, it also has important medico-legal implications. There is a dearth of serious study to document the overall fall from height cases in the metropolis of Karachi or to formulate suggestions to improve the situation. The current study has been undertaken in this direction

Objective: To assess and document the incidence of medico-legally relevant cases of fall from height in the metropolis of Karachi and to suggest ways to minimize this mishap

Methods: Descriptive, retrospective, cross-sectional study of medico-legally relevant cases of falls from height in Karachi registered from 1st January, 2007 to 31st December, 2011; the data having been retrieved from the three medico-legal sections of Karachi located at Abbasi Shaheed Hospital, Jinnah Post-graduate Medical Centre and Civil Hospital and from the Police Surgeon's Office, Karachi. Findings were expressed in numbers and percentages

Results: The total number of registered medico-legal cases of fall from height in Karachi during the study period was 1029. Of these 49.17% [n=506] were reported at ASH, followed by 28.67% [n=295] at CHK and 22.16% [n=228] at JPMC. Of the total number of fall cases, 86.01% [n=885] were non-fatal and 13.99 [n=144] proved fatal. Further, 51.41% [n=455] of the non-fatal cases were referred to ASH while 28.59% [n=253] went to CHK and 20% [n=177] to JPMC. Most of the referrals to ASH were due to location of the hospital

Conclusion: Fall from height is one of the leading causes of medico-legal cases in Karachi with a high morbidity rate as well as mortality with about 15% of them being fatal in the five year period 2007-2011. Despite the incidence, most of these mishaps are preventable. As majority of these cases involve the construction industry, a multi-pronged team effort is required to improve the situation

Anti-inflammatory activity and qualitative analysis of different extracts of Maytenus obscura (A. Rich.) Cuf. by high performance thin layer chromatography method

<p><b>OBJECTIVE</b>To perform aqueous ethanol soluble fraction (AESF) and dichloromethane extract of aerial parts of Maytenus obscura (A. Rich.) Cuf. using high performance thin layer chromatography (HPTLC) and to test anti-inflammatory activity of these extracts.</p><p><b>METHODS</b>HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software were used. The anti-inflammatory activity was tested by injecting different groups of rats (6 each) with formalin in hind paw and measuring the edema volume before and 1 h later formalin injection. Control group received saline i.p. The extracts treatment was injected i.p. in doses of 100 and 200 mg/kg 1 h before formalin administration. Indomethacin (30 mg/kg) was used as standard.</p><p><b>RESULTS</b>The results of preliminary phytochemical studies confirmed the presence of protein, lipid, carbohydrate, phenol, flavonoid, saponin, triterpenoid, alkaloid and anthraquinone in both extracts. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with the green solvents toluene: ethyacetate: glacial acetic acid (5:3:0.2, v/v/v) as mobile phase. HPTLC finger printing of AESF revealed major eight peaks with Rf values in the range of 0.28 to 0.80 and the dichloromethane revealed major 11 peaks with Rf values in the range of 0.12 to 0.76. The purity of sample was confirmed by comparing the absorption spectra at start, middle and end position of the band. Treatment of rats (i.p.) with AESF and dichloromethane in doses of 100 and 200 mg/kg inhibited singnificantly (P<0.05, n=6) formalin-induced inflammation by 50%, 55.9%, 45.5%, and 51.4%, respectively.</p><p><b>CONCLUSIONS</b>HPTLC finger printing of AESF and dichloromethane of Maytenus obscura revealed eight major spots for alcoholic extracts and nine major spots for dichloromethane extracts. These HPTLC profiles may be of great usefulness in the quality control of herbal products containing these extracts. The anti-inflammatory activity of both extracts also revealed the medicinal importance of these extracts. The plant can be further explored for the isolation of phytoconstituents having anti-inflammatory activity.</p>

Chromatographic finger print analysis of anti-inflammatory active extract fractions of aerial parts of Tribulus terrestris by HPTLC technique

<p><b>OBJECTIVE</b>To develop HPTLC fingerprint profile of anti-inflammatory active extract fractions of Tribulus terrestris (family Zygophyllaceae).</p><p><b>METHODS</b>The anti-inflammatory activity was tested for the methanol and its fractions (chloroform, ethyl acetate, n-butanol and aqueous) and chloroform extract of Tribulus terrestris (aerial parts) by injecting different groups of rats (6 each) with carrageenan in hind paw and measuring the edema volume before and 1, 2 and 3 h after carrageenan injection. Control group received saline i.p. The extracts treatment was injected i.p. in doses of 200 mg/kg 1 h before carrageenan administration. Indomethacin (30 mg/kg) was used as standard. HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software for the active fractions of chloroform fraction of methanol extract.</p><p><b>RESULTS</b>The methanol extract showed good antiedematous effect with percentage of inhibition more than 72%, indicating its ability to inhibit the inflammatory mediators. The methanol extract was re-dissolved in 100 mL of distilled water and fractionated with chloroform, ethyl acetate and n-butanol. The four fractions (chloroform, ethyl acetate, n-butanol and aqueous) were subjected to anti-inflammatory activity. Chloroform fraction showed good anti-inflammatory activity at dose of 200 mg/kg. Chloroform fraction was then subjected to normal phase silica gel column chromatography and eluted with petroleum ether-chloroform, chloroform-ethyl acetate mixtures of increasing polarity which produced 15 fractions (F1-F15). Only fractions F1, F2, F4, F5, F7, F9, F11 and F14 were found to be active, hence these were analyzed with HPTLC to develop their finger print profile. These fractions showed different spots with different Rf values.</p><p><b>CONCLUSIONS</b>The different chloroform fractions F1, F2, F4, F5, F7, F9, F11 and F14 revealed 4, 7, 7, 8, 9, 7, 7 and 6 major spots, respectively. The results obtained in this experiment strongly support and validate the traditional uses of this Sudanese medicinal plant.</p>

Theoretical and experimental study on lipophilicity and wound healing activity of ginger compounds

<p><b>OBJECTIVE</b>To correlate the chromatographic and computational method to calculate lipophilicity of selected ginger compounds and to observe the effects of log P on wound healing.</p><p><b>METHODS</b>Mixtures of acetonitrile and water with acetonitrile content between 95% and 50% v/v in 5% increments were kept separately in 10 different chromatographic chambers, saturated with solvent for 2 h. Spots were observed under UV light at λ=254 nm p-anisaldehyde used as a spraying reagent. Theoretical calculation was done using the Alogps 2.1 online program at www.vcclab.org/lab/alogps. For percentage wound contraction, five groups of animal (mice) (25-30 g) of either sex were selected. Wound were created on dorsal surface of animals using toothed forceps, scalpel and pointed scissors. The wound areas were calculated using vernier caliper. After making wound mice were orally administered 35 mg/kg 6-shogoal, 6-gingerol, 8-gingerol and 10-gingerol respectively. Group E as the control group received tap water.</p><p><b>RESULTS</b>The lipophilicity values determined in thin layer chromatography were correlated with the theoretically calculated various log P by linear regression analysis. Significant correlations were found between log P values calculated by software program and the experimental reversed-phase thin-layer chromatography data. Order of wound healing property of ginger compounds is directly dependent on lipophilicity i.e. more lipophilic compound has highest activity.</p><p><b>CONCLUSIONS</b>Experimentally determined lipophilicity (R MO) values were correlated with log P determined by software's and found satisfactory. Lipophilicity (R MO) is a useful parameter for the determination and prediction of biological activity of ginger compounds.</p>