Chemometrics-assisted spectroscopic methods for rapid analysis of combined anti-malarial tablets.

Combination of piperaquine (PQ) (320mg) and dihydroartemisinin (DHA) (40 mg) is an anti-malarial formulation, which is recommended by World Health Organization (WHO). Simultaneous analysis of PQ and DHA can be problematic due to the lack of chromophores or fluorophores in DHA molecule. Whereas PQ possesses strong UV absorption and it presents in 8 times of DHA contents in the formulation. In this study, two spectroscopic methods, Fourier transform infrared (FTIR) and Raman spectroscopy, were developed for the determination of both drugs in combined tablets. The FTIR and Raman spectra were recorded in the attenuate total reflectance (ATR) and scattering modes, respectively. The original and pretreated spectra from FTIR and handheld-Raman were subjected to Unscrambler® program to construct partial least squares regression (PLSR) model comparing with references values obtained from high performance liquid chromatography (HPLC)-UV method. The optimal PLSR models of PQ and DHA from FTIR spectroscopy were obtained from orthogonal signal correction (OSC) pretreatment at the wavenumbers 400-1,800 cm-1 and 1,400-4,000 cm-1, respectively. For Raman spectroscopy of PQ and DHA, the optimal PLSR models were obtained from standard normal variate (SNV) pretreatment at the wavenumbers 1,200-2,300 cm-1 and OSC pretreatment at the wavenumber 400-2,300 cm-1, respectively. Determination of PQ and DHA in tablets from the optimum model was compared with HPLC-UV method. Results were not significantly different at 95% confidence limit (p-value >0.05). The chemometrics-assisted spectroscopic methods were fast (1-3 min), economical and less labor intensive. Moreover, the handheld Raman spectrometer is portable and can be utilized for onsite analysis to facilitate the detection of counterfeit or substandard drugs at ports of entry.

Portable near-infrared and Raman spectroscopic devices as complementary tools for instantaneous quality control of turmeric powder.

INTRODUCTION: Process analytical technology (PAT) guidance is implemented in the quality assurance of phytocompounds to achieve the Industry 4.0 concept. Near-infrared (NIR) and Raman spectroscopies are feasible for rapid, reliable quantitative analysis through transparent packaging without removing the samples from their original containers. These instruments can serve PAT guidance. OBJECTIVE: This study aimed to develop online portable NIR and Raman spectroscopic methods for quantifying total curcuminoids in turmeric samples through a plastic bag. The method mimicked an in-line measurement mode in PAT compared with placing samples into a glass vessel (at-line mode). MATERIALS AND METHODS: Sixty-three curcuminoid standard-spiked samples were prepared. Then, 15 samples were randomly selected as fixed validation samples, and 40 of the 48 remaining samples were chosen as calibration set. The results obtained from the partial least square regression (PLSR) models constructed by using the spectra acquired from NIR and Raman were compared with the reference values from high-performance liquid chromatography (HPLC). RESULTS: The optimum PLSR model of at-line Raman was achieved with three latent variables and a root mean square error of prediction (RMSEP) of 0.46. Meanwhile, the PLSR model of at-line NIR with one latent variable offered an RMSEP of 0.43. For the in-line mode, PLSR models created from Raman and NIR spectra had one latent variable with RMSEP of 0.49 and 0.42, respectively. The R2 values for prediction were 0.88-0.92. CONCLUSION: The models established from the spectra from portable NIR and Raman spectroscopic devices with the appropriate spectral pretreatments allowed the determination of total curcuminoid contents through plastic bag.

A Comparative Study of Benchtop and Portable NIR and Raman Spectroscopic Methods for the Quantitative Determination of Curcuminoids in Turmeric Powder.

Turmeric consumption is continually increasing worldwide. Curcuminoids are major active constituents in turmeric and are associated with numerous health benefits. A combination of spectroscopic methods and chemometrics shows the suitability of turmeric for food quality control due to advantages such as speed, versatility, portability, and no need for sample preparation. Five calibration models to quantify curcuminoids in turmeric were proposed using benchtop and portable devices. The most remarkable results showed that Raman and NIR calibration models present an excellent performance reporting RMSEP of 0.44% w/w and 0.41% w/w, respectively. In addition, the five proposed methods (FT-IR, Raman, and NIR) were compared in terms of precision and accuracy. The results showed that benchtop and portable methods were in good agreement and that there are no significant differences between them. This study aims to foster the use of portable devices for food quality control in situ by demonstrating their suitability for the purpose.

Non-Destructive Analysis of Chlorpheniramine Maleate Tablets and Granules by Chemometrics-Assisted Attenuated Total Reflectance Infrared Spectroscopy.

Non-destructive analysis of chlorpheniramine maleate (CPM), pharmaceutical tablets, and granules was conducted by chemometrics-assisted attenuated total reflectance infrared spectroscopy (ATR-IR). For tablets, an optimum PLSR model with eight latent factors was obtained from area-normalized and standard normal variate (SNV) pretreated ATR-IR spectral data with correlation coefficients (R2) of calibration and cross-validation of 0.9716 and 0.9602, respectively. The model capability for the 42 test set samples was proven with R2 between the reference and model prediction values of 0.9632, and a root-mean-square error of prediction (RMSEP) of 1.7786. The successive PLSR model for granules was constructed from SNV and first derivative pretreated ATR-IR spectral data with two latent factors and correlation coefficients (R2) of calibration and cross-validation of 0.9577 and 0.9450, respectively.

Determination of Chinese hamster ovary (CHO) cell densities and antibody titers from small volumes of cell culture supernatants using multivariate analysis and partial least squares regression of UV-Vis spectra.

Antibody titer and viable cell density (VCD) are two important parameters that need to be closely monitored during the process of cell line development and manufacturing of therapeutic antibodies. Typically, determination of each parameter requires 10-100 µL of supernatant sample, which is not suitable for small scale cultivation. In this study, we demonstrated that as low as 2 µL of culture supernatants were sufficient for the analysis using UV-Vis spectrum assisted with partial least squares (PLS) model. The results indicated that the optimal PLS models could be used to predict antibody titer and VCD with the linear relationship between reference values and predicted values at R2 values ranging from 0.8 to > 0.9 in supernatant samples obtained from four different single clones and in polyclones that were cultured in various selection stringencies. Then, the percentage of cell viability and productivity were predicted from a set of samples of polyclones. The results indicated that while all predicted % cell viability were very similar to the actual value at RSEP value of 6.7 and R2 of 0.8908, the predicted productivity from 14 of 18 samples were closed to the reference measurements at RSEP value of 22.4 and R2 of 0.8522. These results indicated that UV-Vis combined with PLS has potential to be used for monitoring antibody titer, VCD, and % cell viability for both online and off-line therapeutic production process. The process of multivariate analysis and partial least squares regression of UV-Vis spectrum for the determination of CHO cell densities and antibody titers obtained from small volume of cell culture supernatant samples.

Development and Characterization of Stingless Bee Propolis Properties for the Development of Solid Lipid Nanoparticles for Loading Lipophilic Substances.

Stingless bees are insects which are popularly bred by agriculturists in the eastern region of Thailand for the pollination of their orchards. The products from stingless bee breeding include bee honey and bee propolis. The objective of this experiment is to study the possibility of developing stingless bee propolis wax into solid lipid nanoparticles (SLN) by the comparison of five surfactants (Brij 721, Cremophor WO 7, Myrj 52, Poloxamer 188, and Tween 80). Each surfactant is used at three concentrations: 10%, 20%, and 30%. A master formula is selected according to the following: physical features, particle size, zeta potential, and entrapment. The results showed that Brij 721 and Myri 52 at 20% can be used in preparing SLN and have good preservation properties for vitamin E (size: 451.2 nm and 416.8 nm, zeta potential: - 24.0 and - 32.7; % EE: 92.32% and 92.00%, resp.). Therefore, they are further developed by adding the following drugs at low solubility: curcumin, ibuprofen, and astaxanthin. It is found that a formula using the surfactants Brij 721 and Myrj 52 at 20% have similar drug entrapment. The entrapment study involves curcumin 82%, ibuprofen 40%, and astaxanthin 67%. Moreover, the cytotoxicity test of blank solid lipid nanoparticle found no toxicty in fibroblast cell line (CRL-2522). Therefore, from this study, it is determined that stingless bee propolis wax has the potential to be developed to provide more efficient SLN in the future.

Antioxidant Effects and in vitro Cytotoxicity on Human Cancer Cell Lines of Flavonoid-Rich Flamboyant (Delonix regia (Bojer) Raf.) Flower Extract.

BACKGROUND: Cancer is a non-communicable disease with increasing incidence and mortality rates worldwide, including Thailand. Its apparent lack of effective treatments is posing challenging public health issues. INTRODUCTION: Encouraging research results indicating probable anti-cancer properties of the Delonix regia flower Extract (DRE) have prompted us to evaluate the feasibility of developing a type of product for future cancer prevention or treatment. METHODS AND RESULTS: In the present report, using High-Performance Liquid Chromatography (HPLC), we demonstrate in the DRE the presence of high concentrations of three identifiable flavonoids, namely rutin 4.15±0.30% w/w, isoquercitrin 3.04±0.02 %w/w, and myricetin 2.61±0.01% w/w, respectively, while the IC50 of DPPH and ABTS assay antioxidation activity was 66.88±6.30 µg/ml and 53.65±7.24 µg/ml, respectively. DISCUSSION: Our cancer cell line studies using the MTT assay demonstrated DREs potent and dosedependent inhibition of murine leukemia cell line (P-388: 35.28±4.07% of cell viability remaining), as well as of human breast adenocarcinoma (MCF-7), human cervical carcinoma (HeLa), human oral cavity carcinoma (KB), and human colon carcinoma (HT-29) cell lines in that order of magnitude. CONCLUSION: Three identifiable flavonoids (rutin, isoquercitrin and myricetin) with high antioxidation activity and potent and dose-dependent inhibition of murine leukemia cell line and five other cancer cell lines were documented in the DRE. The extract's lack of cytotoxicity in 3 normal cell lines is a rare advantage not usually seen in current antineoplastic agents. Yet another challenge of the DRE was its low dissolution rate and long-term storage stability, issues to be resolved before a future product can be formulated.

Simple and fast fabrication of microfluidic paper-based analytical device by contact stamping for multiple-point standard addition assay: Application to direct analysis of urinary creatinine.

In this work, a microfluidic paper-based analytical device (µPAD) for simultaneous multiple-point standard addition assay was fabricated by rubber stamping the hydrophobic barrier pattern onto a laboratory filter paper. The µPAD has a central zone from which an applied sample flows into eight surrounding narrow channels to which had been added the standard solutions. Each channel is connected to a circular area loaded with the reagent. The opposite end of this reagent zone is connected by second narrow channel to the final circular detection zone. The µPAD was applied to the measurement of creatinine in human urine. After addition of a urine sample, the orange-colored product arising from the Jaffé reaction is formed at the eight detection zones. A digital image of the µPAD is then recorded and the ratio of the red/green (R/G) intensity obtained using the ImageJ™ program is used in the quantitation of creatinine. The normal standard addition calibration line is constructed using the intensity ratio against the added creatinine concentrations (50-1000 mg L-1). Good linearity was achieved (r2 ˃ 0.99). There were no significant differences between the creatinine content using an HPLC method (paired t-test at 95% confidence, tstat = 1.78, tcritical = 2.26, n = 10). The use of simultaneous multiple-point standard addition calibration allows rapid determination of creatinine in urine with elimination of matrix interference.