Authors:Lensi Kumssa, Thomas Layloff, Ariaya Hymete, and Ayenew Ashenef
According to World Health Organization (WHO) 10% of the medicines in the Low and Middle Income Countries (LMICs) are of poor quality posing a major public health threat. One way to circumvent such problem is the development and deployment of rapid, economical and efficient analytical methods. Hence this research aims to develop a High-Performance Thin Layer Chromatography (HPTLC) method for the determination of doxycycline hyclate. A rapid and simple HPTLC method with densitometry detection at 360 nm to determine doxycycline hyclate in capsules and tablet formulations was developed and validated. HPTLC was performed on glass plates coated with C18 reverse phase silica gel 60 F254 and pretreated with 0.27 M ethylenediaminetetraaceticacid (EDTA) solution. The mobile phase was dichloromethane: methanol: acetonitrile: 1% aqueous ammonia in the ratio of 10:22:53:15 (v/v). The linearity range lies between 200 and 1,000 ng/spot with correlation coefficient of 0.997. The Rf value is 0.5 ± 0.02%. Recoveries were in the range of 94.50–100.5%. Limit of detection and limit of quantitation values for doxycycline hyclate were 40 and 160 ng/spot respectively. The developed method was validated as per ICH guidelines. Thus, it was found to be accurate, precise, specific and robust. In forced degradation study, doxycycline hyclate was found to degrade in acidic and alkaline media, and through oxidative stress. The drug was found to be relatively stable to heat and photo degradation. The method was successfully applied for the routine quantitative analysis of dosage forms containing doxycycline hyclate. The developed method offered comparable results (as confirmed by F-test) with that of the HPLC pharmacopoeial (BP) analysis method.
Authors:Wael Alshitari, Fatimah Al-Shehri, Deia Abd El-Hady, and Hassan M. Albishri
Statins drugs are thought to be among the most prescribed drugs worldwide for the treatment of hypercholesterolaemia. A simple and reliable RP-HPLC method has been successfully employed for simultaneously separating and qualifying three statin drugs including atorvastatin, rosuvastatin and simvastatin in pharmaceutical tablets. The optimal conditions were mobile phase 50:50 (v/v) (formic acid pH 2.50: ETOH), column temperature 40.00 °C, detection wavelength 238.00 nm, and flow rate 1.00 mL/min. The proposed method has been validated based on the ICH guidelines in terms of linearity, precision, accuracy, and limit of detection and limit of quantification. The linear range investigated 2.0–80.0, 4.0–100.00, and 12.00–120.00 µg/mL for rosuvastatin, atorvastatin and simvastatin respectively with coefficients of determination (R2) within the range of 0.9993–0.9995. The LOD and LOQ for rosuvastatin, atorvastatin and simvastatin were (1.57, 4.76 µg/mL), (1.87, 5.66 µg/mL), (3.46, 10.49 µg/mL) respectively. In addition, in order to evaluate the feasibility of the method developed, it was employed towards the quantification of the pharmaceutical tablets for the analytes investigated and excellent recovery was obtained.
Authors:Hisham Elrefay, Omnia A. Ismaiel, Wafaa S. Hassan, Abdalla Shalaby, and Ali Fouad
A new, sensitive, stability-indicating reversed-phase HPLC method was validated and applied for the simultaneous quantitation of sodium valproate and two paraben preservatives; methylparaben, and propylparaben in the liquid dosage form. Stability tests were carried out through exposure of the analyte's solution to stress conditions. Separation of the analytes was achieved on (waters) C18 Column (150 mm × 3.9 mm, 5 μm). A mixture of 0.05 M monobasic potassium phosphate pH 3.5 and acetonitrile (50:50; v/v) was applied at 1.5 ml/min flow rate and UV detection wavelength at 210 nm. The degradation products and the analytes were completely separated. The linearity was performed in the range of 50–150 % from a target concentration of 10 μg/ml propylparaben, 90 μg/ml methylparaben, and 2.88 mg/ml sodium valproate with a coefficient correlation (R2) 1.0 for methylparaben, propylparaben and sodium valproate. The validation results of the suggested method were in a good agreement with ICH guidelines. Application of the proposed method for analysis of liquid dosage forms was successfully carried out in the routine quality control process.
Authors:Mohammed Hamed Alqarnı, Mohamad Ayman Salkini, Prawez Alam, Mazen Talal Alanazı, Maged Saad Abdel-Kader, and Samah M. El Sohafy
Plants secondary metabolites undergoes qualitative and quantitative variation due to environmental and growth factors. It is a crucial factor to select the proper time for collection of medicinal plants to assure maximum content of active components reflected as maximum efficacy. Olive leaves (Olea europaea L.) are known traditionally for their antidiabetic effect. The secoiridoid glycoside oleuropein is the main active component of Olive leaves responsible for the biological activity. The current study was conducted to monitor the seasonal variation of oleuropein in Olives leaves collected from the same location. To achieve this goal a validated HPLC method following the ICH guidelines was established. Separation was conducted using RP18 column and a mobile phase consisted of ultrapure water containing 20% acetonitrile and 1% acetic acid. Detection was performed at 254 nm with 1 mL/min flow rate. The method was simple, linear, accurate, precise, specific and robust. The analyses revealed considerable variations in the level of oleuropein throughout the year. This variation cannot be explained by temperature variation during the year. Two points of high levels of oleuropein were detected prior to flowering stage and ripening of the fruits. The levels of growth regulators most likely is responsible for the increased production of oleuropein. It is recommended that leaves intended for medicinal use to be collected during the fruiting stage prior to fruit ripening.
Spiny coriander (Eryngium foetidum L.) is a perennial medicinal herb grown in the tropical regions worldwide. In India, it is used as a potential spice for garnishing and flavoring the dishes and treating several ailments. Eryngium spp. found in coastal Odisha, India has a strong aroma similar to the seasonal Coriandrum. The volatile flavor constituents of the unique plants were analyzed through headspace solid-phase microextraction (HS-SPME) using capillary gas chromatography (GC) and gas chromatography-tandem mass spectrometry (GC–MS/MS). The volatile compounds exhibited high chemodiversity, with 10-undecenal as the major component in leaves (44.98%) and branches (57.43%). Fourier-transform infrared (FTIR) spectroscopy identified eight major peaks grouped into six main regions. Chemo profiles of these two corianders were overlapped and showed similar area differences in the spectral peak. The lesser-known perennial Eryngium with high chemodiversity would be a better alternative to the seasonal coriander for aromatic, pharmaceutical, and industrial uses.
Authors:Pengfei Li, Xi Zhang, Shumin Wang, Jing Wang, Ziqi Liu, Chen Wang, Weihang Tong, and Lihong Liu
Letrozole is one of the third generation aromatase inhibitors. It is suitable for the treatment of postmenopausal patients with advanced breast cancer and early treatment of breast cancer. It is necessary to develop a rapid, reliable, selective and sensitive LC–MS/MS assay to determine letrozole in human plasma to evaluate the clinical efficacy and adverse reactions with clinical pharmacokinetic and therapeutic drug monitoring. Separation was carried out on a Kromasil-C18 column using acetonitrile-water (55: 45, v/v) as mobile phase. Detection was carried out by multiple reaction monitoring on a 3200Qtrap mass spectrometry. The method needed one-step protein precipitation procedure only, and the cycle time was 2.5 min allowing 500–550 samples per day. It was linear within 0.30–50.00 ng/mL for plasma with the limit of detection (LOD) of 0.030 ng/mL. The intra- and inter-day RSD were 5.51–8.63%, 2.28–9.95% and the RE was 0.18–1.65%. The recovery rates of letrozole and internal standard for plasma were 89.30–98.55%. Letrozole was stable under all the conditions in the study. The method was sensitive enough to quantitate letrozole over a period of 288 h after giving a single oral dose of 2.5 mg–24 Chinese healthy volunteers. The absorption of letrozole was rapid with small individual difference, the tissue distribution of letrozole was more than that in blood, and the clearance was slow. Letrozole was similar to three-compartment model in vivo. Due to metabolism and excretion, the AUCs of letrozole varied greatly among individuals.
Authors:Tóth József Attila, Döbröntey Réka, Szegi Tamás, Michéli Erika, and Csorba Ádám
Szervesszén térképezést segítő módszertani kutatásként vizsgáltuk egy szántóföldi művelés alatt álló terület, 3 mélységből származó mintáinak MIR reflektanciáját, illetve szervesszén tartalmát (Walkley-Black). Ezt követően a spektroszkópia mérések eredményeit használtuk a talaj szervesszén-mennyiségének (TOC %) becslésére. Tettük ezt 3 mintakijelölési módszer (Kennard-Stone Sampling - KSS, K-means Sampling - KMS, Latin Hypercube Sampling - LHS) bevonásával, az így kijelölt kalibrációs mintákkal a PLSR modell segítségével becslést végeztünk az adathalmaz további értékeire. Annak érdekében, hogy tábla szintű szervesszén meghatározás során teszteljük becslési pontosságukat, a modellek reprezentativitását – különböző validációs/kalibrációs arány esetén – statisztikai mutatókkal (R2, RMSE) ellenőriztük.
Az eredményekben részleteiben vizsgáltuk a különböző becslési modellek reakcióját eltérő arányú kalibráció és validáció esetén. A modellek R2 és RMSE értékei alapján kijelöltük, hogy mely modellek működtek pontosan még alacsony kalibráció esetén is, illetve abszolút értelemben véve melyik modell volt leghatékonyabb.
Az összehasonlítás eredményeként kijelenthető, hogy az általunk vizsgált talajkörülmények között a 30% alatti, valamint a 70% feletti mintaszámú kalibráció a mintakijelölési módszerek megbízhatóságának ingadozását eredményezte. Az összes minta 30%-val történő kalibráció esetén legjobb eredményt a KSS adta, így ez tekinthető a leggazdaságosabb módszernek. Az abszolút értékben vett legkisebb hibát a K-means sampling eredményezte, a minták 90%-val történő kalibrációt követően.
Kijelenthető, hogy az alkalmazott módszertan esetünkben alkalmas volt – a reprezentativitás megtartása mellett – a szükséges minták számának, ergo a táblaszintű szervesszén-felmérés költségeinek csökkentésére. Továbbá a mintakijelölési módszerek becslési hatékonyságának összehasonlítására is megfelelt az általunk alkalmazott statisztikai vizsgálat. A módszertan a jövőben kiinduló alapja lehet hasonló jellegű kutatásoknak, valamint tábla szintű szervesszéntérképek elkészítésének. A szélesebb körű alkalmazást megelőzően a modelleket nagyobb varianciájú adathalmazok esetén is tesztelni szükséges.
Within the framework of the present research, we mapped the organic carbon content of an arable area, during which we measured the MIR reflectance and organic carbon content (Walkley-Black) of the soil samples collected from the area at three different depths. Subsequently, the results of spectroscopic measurements were used to improve the estimation of the soil organic carbon content (TOC %). Three sample selection models were involved (KSS, KMS, LHS), and with the selected calibration samples, we estimated the additional values of the data set using the PLSR model. In order to test the accuracy of estimation for a table-level organic carbon determination, the representativeness level of the models was checked with statistical indicators (R2, RMSE) at different validation / calibration ratios.
In the results, we thoroughly examined the response of different estimation models with different ratios of calibration and validation. Based on the R2 and RMSE values of the models, we determined which models worked precisely even at low calibration, and in absolute terms, which model was the most efficient.
As a result of the comparison, it can be stated that under the soil conditions we examined, calibration with a sample number below 30% and above 70% caused significant fluctuations in the reliability of the sampling methods. Kennard-Stone sampling (KSS) gave the most precise results for calibration with 30% of all samples, thus it is considered the most economical method. The smallest error overall was given by K-means sampling after calibration of 90% of the samples.
It can be stated that the methodology used in this study was suitable to reduce the samples required for analysis - while maintaining representativeness - therefore reducing the costs of the field-level organic carbon survey.
Furthermore, the statistical analysis we used to compare the estimation efficiency of the sampling methods was also appropriate. The methodology we use may be the basis for similar research in the future, as well as for the production of table-level organic carbon maps. Prior to wider application, models also need to be tested for higher variance datasets.