Quality Assessment and Origin Identification on Four Species of Curcumae Radix by HPLC Assay and Fingerprint Analysis

BackgroundCurcumae Radix is a multi-source Chinese medicine called Yujin. It is derived from the root tubers of four Curcuma species including Curcuma wenyujin , C. kwangsiensis , C. phaeocaulis and C. longa. The identification of the root tubers of the four Curcuma species often caused confusions. MethodsIn this study, we developed HPLC/PDA methods to differentiate these four species of Curcumae Radix by HPLC assay and fingerprint analysis. The methods developed were also validated with their precision, repeatability and accuracy. ResultsCurdione could only be detected in C. wenyujin . For C. kwangsiensis , C. phaeocaulis , if the relative peak area value of peak 1 and germacrone peak of the samples was more than 1.024, it could be identified as C. phaeocaulis of Curcumae Radix. Curcumin, desmethoxycurcumin and bisdesmethoxycurcumin could only be detected in C. longa . We also developed a simple method to determine curcumin, desmethoxycurcumin and bisdesmethoxycurcumin in C. lon ga and germacrone in the other three species of Curcumae Radix. ConclusionsBecause of the definite difference of main ingredients between curcumin-free group (C. wenyujin , C. kwangsiensis and C. phaeocaulis ) and curcumin containing C. longa from this study, four Curcuma species of Curcumae Radix in the pharmacopeia should be separated into two monographs, with curcumin-free and germacrone containing species as one and curcumin containing C. longa as the second monograph. The HPLC/PDA methods of HPLC assay and fingerprint analysis developed in this study could be used for the quality control of these four species of Curcumae Radix. Keywords: Curcumae Radix, HPLC, assay, fingerprint, curcumin, desmethoxycurcumin, bisdesmethoxycurcumin, germacrone, curdione

Stability-indicating HPLC method to determine the stability of extemporaneously prepared vancomycin oral solution cups

Purpose To determine the stability of compounded sweetened vancomycin oral formulations in plastic unit dose cups stored up to 180 days under 2 temperature conditions: refrigeration (2°C-6°C) and room temperature (25°C with 60% relative humidity). Methodology A stability-indicating high-performance liquid chromatography (HPLC) method was developed to analyze vancomycin in the presence of degradation peaks. The stability of extemporaneously compounded vancomycin solution stored in oral unit dose cups was investigated using this method. The tested vancomycin oral solutions were compounded formulations of 125 mg/2.5 mL and 500 mg/10 mL. Three oral unit dose cups from each storage condition were withdrawn and assessed for stability on days 0, 3, 7, 15, 22, 30, 90, 120, 150, and 180 as per United States Pharmacopeia guidelines. The assay of vancomycin was carried out by using a calibrated stability-indicating HPLC method. Results The stability-indicating HPLC assay showed that vancomycin completely degraded within 2 hours when exposed to highly acidic or basic pH conditions. No precipitation, cloudiness, or color changes were observed during the study under either temperature condition. The HPLC assay revealed that vancomycin oral solution cups retained greater than 90% of the initial concentrations of vancomycin for 30 days when stored at room temperature (25°C and 60% relative humidity) and for 180 days with refrigeration (2°C-6°C). Conclusion Vancomycin oral formulations were stable for long-term storage periods beyond those specified in manufacture guidelines. Our data suggests the extended stability of vancomycin oral solutions compounded for hospital use can be extended.

Variability of the arbutin content in wild growing populations of Arctostaphylos uva-ursi (L.) Spreng from Korab Mountain, Western Balkan

Due to the presence of arbutin and hydroquinone in the bearberry plant (Arctostaphylos uva-ursi (L.) Spreng., Ericaceae), it is widely used as a urinary tract antiseptic and diuretic. The herbal substance consists of whole or cut dried leaves, and it should not contain less than 7.00% of anhydrous arbutin (Ph.Eur.10). The supply of this herbal substance in Balkan countries mainly comes from a wild plant harvested in the mountains. It is a common practice to collect the leaves during the flowering season of the plant (June-July). There is an abundance of wild growing natural populations of bearberry on Korab Mountain that represents a unique natural resource of this herbal substance for both countries, Albania and North Macedonia. The aim of the present study was to determine the arbutin content and to assess its variability in the bearberry leaves of seven wild populations from Korab Mountain. The leaves were collected monthly (May-October) during a period of 3 years (2014-2016). The HPLC assay (Ph.Eur.10) revealed that all populations have arbutin content over 7.00% (7.03-9.42%) and that the highest content of arbutin can be attained in September/October, during the phase after the fructification. Statistical analysis showed that there is significant difference between the content of arbutin in the different populations. The content was related to the altitude of the collection site and the collection month and year.

Development of a Comprehensive HPLC Method for Determination of Product-related Impurities and Assay of Active Ingredients in Papaverine Hydrochloride Products

Papaverine hydrochloride products are used as anticonvulsants in routine medical practice. Most of the approved product specification files include thin-layer chromatography for assessment of product-related impurities and UV spectrophotometry for determination of active pharmaceutical ingredients. An HPLC assay is not used for determination of papaverine hydrochloride in drug dosage forms.The aim of the study was to develop an HPLC test method for determination of product-related impurities and for quantification of papaverine hydrochloride in solutions for injection, tablets, and rectal suppositories.Materials and methods: samples of the following Russian-made papaverine products were used in the study: Papaverine, solution for injection, 20 mg/mL; Papaverine, rectal suppositories, 20 mg; Papaverine, tablets, 40 mg. The Agilent 1260 Infinity II DAD System was used for the HPLC assay, and the Agilent 8453Е UV-Vis System was used for recording UV spectra. The determination of product-related impurities and the assay of active ingredients were performed simultaneously by HPLC using a reversed-phase column Kromasil 100-5-C18, 250×4.6 mm, 5 μm, the gradient elution mode, and detection at 238 nm. Papaverine Hydrochloride USP RS, 99% purity, and Noscapine EP CRS were used as reference standards.Results: the study demonstrated that determination of product-related impurities and assay of active ingredients in papaverine products can be performed simultaneously using HPLC.Conclusions: the authors proposed an HPLC test method for determination of active ingredients in papaverine products, which is aligned with the “consistent standardisation” principle and can be recommended for inclusion into draft monographs for papaverine products.

A Thorough HPLC Assay of Quaternary Veterinary Formulation coupled with Environmental Assessment Tool

A new and accurate reversed phase HPLC method with UV detection has been established for any veterinarian analyst for simultaneous determination of a veterinary quaternary mixture of sulphadimidine sodium (SDS), sulphaquinoxaline sodium (SQS), diaveridine (DVD) and vitamin K3 (VTK3) in their formulation. The stationary phase was SEA C18 column (250 × 4.6 mm i.d., 5 μm particle size) at 25°C with an isocratic mode, using a mobile phase containing a mixture of methanol:acetonitrile:distilled water in the ratio of (20:20:60, by volume). The flow rate was 0.8 mL min−1, and UV detection was performed at 230 nm. The HPLC assay was coupled with Environmental Assessment Tool (EAT), which represents a simple and proficient approach for profiling the greenness of the method. This takes into consideration the environmental, health and safety issues for all solvents that involved in the chromatographic method and calculates a total score that can be used for comparison of the greenness of different methods. The method was found to be linear over (0.5–30) μg/mL for all cited drugs with mean percentage recoveries (99.56 ± 1.141) for VTK3, (99.56 ± 1.056) for DVD, (99.62 ± 1.482) for SDS and (99.52 ± 1.205) for SQS. The results were statistically compared with those of the official and reported methods; using Student’s t-test and F-test, showing no significant difference with respect to accuracy. Specificity of the applied method was assessed by analyzing the laboratory-prepared mixtures. The developed method was validated according to ICH guidelines. The proposed methodology can be applied for rapid routine assay of this combination.

A sensitive HPLC-FLD method for the quantification of 6-O-demethylmenisporphine isolated from Menispermi Rhizoma in rat plasma

Background To investigate the pharmacokinetics of 6-O-demethylmenisporphine, an oxoisoaporphine alkaloid with significant anti-tumor activities and isolated from Menispermi Rhizoma, a novel and sensitive HPLC assay was established for 6-O-demethylmenisporphine quantification in rat plasma. Methods Peak responses were detected by a highly selective and sensitive fluorescence detector with 426-nm excitation and 514-nm emission wavelengths. Curcumin was employed as the internal standard (IS). A Capcell Pak C18 column (150 mm × 4.6 mm i.d., 5 μm) and an isocratic elution procedure with a flow rate of 1.0 mL/min were used to exclude the endogenous interfering substance. Acetonitrile-water (68:32, v/v) containing 1% formic acid was employed as mobile phase. A 7-point calibration curve that covered the concentration range of 10–2500 ng/mL was constructed. Results A good linearity was observed with a correlation coefficient (r) of 0.9993. The lower limit of quantification for 6-O-demethylmenisporphine was 10 ng/mL. The mean recoveries of analyte in rat plasma exceeded 80.5%. The precision at four concentration levels was within 11.3% and the accuracy ranged from − 7.6 to 6.7%. Conclusion Using this new HPLC-FLD method, the investigation of plasma samples from rats following oral dosing of neat compound and Menispermi Rhizoma extract was successfully conducted. The results will provide a reference for the evaluation of preclinical safety of 6-O-demethylmenisporphine.