Long-acting injectable formulations are preferred over conventional formulations for the treatment of chronic diseases. The regulatory guidelines and pharmacopoeia have remained silent on dissolution methods for long-acting injectable formulations due to their diverse nature. The lack of compendial method for dissolution testing increases the duration of approval process for long-acting injectable formulations. This article reviews various dissolution methods used to study in vitro drug release profile of long-acting injectable formulations. Compendial as well as noncompendial methods, such as-flow-through cell method, sample and separate method, the dialysis method are used by researchers for drug release profile of long-acting injectable formulations. This review article also highlights the advantages and disadvantages of reported dissolution methods. The compiled work will help the researchers in designing the bio-relevant dissolution method and expedite the development of long-acting injectable formulations.

Objective: To establish an HPLC method of the content and related substances of compound amino acid injection(3AA). Methods: RP-HPLC was adopted to determine compound amino acid injection(3AA), combining with the use of two-dimensional column switching-LC/MSⁿ method was applied to separate and identify the impurities. The determination was performed on Capcell PAK AQ C₁₈(250 mm×4.6 mm, 3 μm) column with 0.2 mol·L^-1 sodium dihydrogen phosphate solution (adjusted pH to 2.8 with phosphoric acid) -acetonitrile (98∶2) as mobile phase at the flow rate of 1.0 mL·min^-1. The column temperature was 40 ℃, and the detection wavelength was 210 nm. And the injection volume was 20 μL. The LC/MSⁿ method was performed on a Thermo Accucore AQ C₁₈ (100 mm×4.6 mm, 2.6 μm) column with 0.1% formic acid solution as mobile phase A, 0.1% formic acid solution-acetonitrile as mobile phase B, at a flow rate of 0.4 mL·min^-1, and at a column temperature of 40 ℃. The mass spectrometry conditions were performed using an ESI ionisation source in the positive-ion scanning mode with a scan range of m/z 100-1 000, and the secondary mass spectrum was carried out by data-dependent scanning. Results: The related substances were completely separated from the main constituents in RP-HPLC. The standard curve of valine was linear over the range of 1.263-5.050 mg·mL^-1, with the average recovery of 99.0% (n=9). The standard curve of isoleucine was linear over the range of 1.350-5.402 mg·mL^-1, with the average recovery of 99.4%(n=9). The standard curve of leucine was linear over the range of 1.647-6.588 mg·mL^-1, with the average recovery of 99.5%(n=9). The main impurities in the three batches of samples were all process impurities introduced from the raw materials, with methionine content of 4.344 μg·mL^-1, 3.751 μg·mL^-1, 4.503 μg·mL^-1, respectively, phenylalanine content of 4.636 μg·mL^-1, 4.889 μg·mL^-1, 4.753 μg·mL^-1, respectively. The maximum single impurity contents were 0.01%, 0.02% and 0.01%, respectively. Conclusion: The method is proved by the methodology validation that it can be used for the quality control of compound amino acid injection(3AA).

This article focuses on the key experimental operations in the study about the applicability of microbial limit testing methods for drugs, summarizing literature and experience from the preparation of test solution, design of methods for microbial counting and control bacteria inspection, operation of microbial recovery test, bacterial liquid concentration counting, and operational methods for controlling bacterial & fungal purity. Emphasis was placed on the preparation methods of water-soluble test samples with strong antibacterial effects, non-oil and fat test samples that were not easily soluble and dispersed, and the test solution for oil and fat test samples. Detailed introduction of the sequential experimental plan for methods of aerobic bacterial count, mold & yeast count, and control bacterial inspection were introduced. Four operational methods for adding bacteria about membrane filtration were explained, and the impact of different ways of adding bacteria on the results of the method was analyzed. Summarized three counting methods for bacterial & fungal concentrations. Detailed sharing of operational experience on controlling the purity of bacterial solution. Summarized the current status about research on the applicability of microbial limit testing methods of drugs. Four suggestions for the future development of research on the applicability of microbial limit testing methods for drugs have been proposed:(1) Unify the operation about bacteria adding method in membrane filtration,scientific and reasonable;(2)Strengthen supervision of pharmaceutical production enterprises and review of microbial limit testing methods for drugs;(3)Unify the microbial limit testing methods for the drugs of the national sampling plan,and gradually collect and bind into a book;(4)Strengthen the research on the applicability of microbial limit testing methods for excipients.

Objective: To establish the substances benchmarks fingerprint of Daqinglong decoction and determine the content and transfer rate of 6 index components, and study the transfer rule of Daqinglong decoction substances benchmarks value, which laid the foundation for the study of Daqinglong decoction compound preparation and quality standard. Methods: Fifteen batches of substances benchmarks of Daqinglong decoction were prepared, and the paste rate was determined. To determine the contents of ephedrine, pseudoephedrine, amygdalin, and cinnamic acid, HPLC method was performed on a Dikma Platisil C₁₈ column (250 mm×4.6 mm, 5 μm). And 0.15% phosphoric acid water (A) and acetonitrile (B) were used as mobile phases with gradient elution at a flow rate of 1 mL·min^-1. The column temperature was 25 ℃, the detection wavelength was 207 nm, and the injection volume was 10 μL. To determine the contents of liquiritin and glycyrrhizic acid, 0.05% phosphoric acid water (A) and acetonitrile (B) were used as mobile phases with gradient elution. The detection wavelength was 237 nm. The transfer rate was calculated, and the transfer analysis from the decoction pieces to the substances benchmarks was carried out. Results: The fingerprint similarities of 15 batches of Daqinglong decoction substances were ≥0.921. Fifteen common peaks were identified, including 8 from Ephedrae Herba, 3 from Cinnamomi Ramulus, 2 from Glycyrrhizae Radix et Rhizoma(fried), and 2 from Armeniacae Semen Amarum(here). The contents of ephedrine, pseudoephedrine, amygdalin, liquiritin, cinnamic acid and glycyrrhizic acid were 0.94%-1.30%, 0.56%-1.02%, 0.70%-1.25%, 0.18%-0.32%, 0.05%-0.10% and 0.46%-1.23%, respectively. The transfer rates were 41.67%-59.47%, 42.66%-59.74%, 17.59%-36.34%, 21.48%-44.75%, 31.06%-48.89% and 12.95%-25.15%, respectively. The paste yields of substances benchmarks were 11.98%-13.38%. Conclusion: The fingerprint combined with the determination of paste rates and index component contents are used to study the quantitative value transfer of Daqinglong decoction substance benchmarks, and initially establish a scientific and stable substance benchmarks quality evaluation method, which can provide reference for the future research on compound preparation.

Objective: To screen the suitable monitoring method applicable to oligotrophs and review the bioburden and microbial species of oligotrophic environments in pharmaceutical water systems and cleanrooms. Methods: The culture conditions and detection methods suitable for microorganisms under oligotrophic conditions were optimized by comparing the counting results of microorganisms in laboratory pure water with R₂A and TSA media at different temperatures, media, and incubation times. In addition, monitoring of oligotrophs in pharmaceutical environments using both the traditional and oligotrophic assays were conducted, combining with the techniques of 16s rDNA sequencing and MALDI-TOF MS for multiphase microbial identification and analysis of the isolated microorganisms under oligotrophic conditions. Results: The colony counting methods, the type of culture media, and the incubation intervals significantly affected microbial enumeration. The isolation method using oligotrophic medium R₂A was more effective than the TSA medium in this study. The profiles of microorganisms isolated from the R₂A and TSA media were unique. The isolation rate of Gram-negative bacteria in the R₂A medium reached 37.0%, however, that of Gram-negative bacteria in the TSA medium was only 14.0%. In addition, several strains of potential pathogens initially isolated from the R₂A medium grew slowly in the TSA medium and were easily missed by the traditional monitoring method. Conclusion: With the development of the pharmaceutical industry, the contamination proportion of human-associated Gram-positive cocci might gradually decrease, the oligotrophic culture method as a powerful supplement to traditional monitoring techniques can help to detect potential objectionable microorganism contamination risks at an early stage.

Objective: To establish an HPLC fingerprint of Zuoyu’an San and conduct quality control by combining chemometrics and quantitative analysis of multi-components with a single-marker(QAMS). Methods: HPLC was used to establish the fingerprint spectra of 12 batches of Zuoyu’an San and perform similarity evaluation. Cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were combined to perform chemical pattern recognition analysis on different batches of Zuoyu’an San. At the same time, QAMS method was established using rhein as the internal standard reference. The relative correction factors between the internal reference and jatrorrhizine, palmatine, berberine, aloe-emodin, emodin, chrysophanol, and physcion were established. The external standard method (ESM) and QAMS method were used to determine jatrorrhizine, palmatine, berberine, aloe-emodin, rhein, emodin, chrysophanol, and physcion in Zuoyu’an San, respectively. To verify the reliability of QAMS metod results, the contents of 8 components of physcion were determined. Results: A total of 23 common peaks were calibrated in the fingerprint, and 8 components were identified by comparing with the mixed reference solution. The similarity between the samples and the control spectrum was between 0.952 and 0.994; CA divided 12 batches of Zuoyu’an San samples into 3 categories; PCA showed that there were certain differences in the chemical composition of different batches of Zuoyu’an San, and extracted 5 principal components that affected the quality evaluation of Zuoyu’an San; OPLS-DA screened out 13 potential marker components (VIP>1) among 23 common peaks that caused quality differences between different batches of Zuoyu’an San samples; There was no significant difference between the QAMS method calculated value and the ESM measured value. Conclusion: The established HPLC fingerprint and multi-index component determination method was accurate and simple, and combined with chemometric methods, could provide reference for the quality control and evaluation of Zuoyu’an San.

Objective: To establish the HPLC characteristic chromatogram for Gujing Maisiha tablets, and evaluate the quality using chemical pattern recognition. Methods: The Welch Xtimate C₁₈ column (250 mm × 4.6 mm, 5 μm) was used and the mobile phases were acetonitrile (A) and 0.1% formic acid (B) in gradient elution (0-10 min,5%A→15%A;10-20 min,15%A;20-40 min,15%A→40%A;40-65 min,40%A→70%A; 65-66 min,70%A→90%A;66-80 min,90%A;80-81 min,90%A→5%A;81-85 min,5%A). The detection wavelength was 254 nm. The column temperature was 35 ℃ and the flow rate was 1.0 mL·min^-1. Gujing Maisiha tablets were analyzed to construct characteristic chromatogram. The quality of Gujing Maisiha tablets was assessed through similarity evaluation, cluster analysis, principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA). Results: The characteristic chromatogram of Gujing Maisiha tablets was established, which contained the twenty-eight distinct peaks, and seven compounds identified as gallic acid, chlorogenic acid, picrocrocin, isoquercitrin, crocin-Ⅰ, crocin-Ⅱ, eugenol. The similarities for the ten batches of Gujing Maisiha tablets were more than 0.98, with no significant difference observed. Cluster analysis segregated the samples into two distinct groups according to the Euclidean distance of 10, which was consistent with the results of PCA analysis. PLS-DA analysis screened out five components from Flos Caryophylli, Rosae Rugosae Flos, Olibanum that may cause differences between samples. Conclusion: The method of HPLC characteristic chromatogram of Gujing Maisiha tablets is effective, feasible and reproducible, which can provide reference for the study of quality standard of this product.

Objective: To establish an innovative analytical method based on high resolution sampling two-dimensional chromatography (HiRes 2D-LC) for determination of the content of vitamin D₃ in vitamin D drops. Methods: Two-dimensional liquid chromatography was used. Thermo HYPERSIL Gold Silica (100 mm×2.1 mm, 1.9 μm) column was used in the first dimension with n-hexane-n-amyl alcohol (996∶4) as mobile phase. The flow rate was 0.2 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. In the second dimension liquid chromatography, ShimPack Velox Hilic (50 mm×2.1 mm, 2.7 μm) was used as the column with n-hexane-n-pentanol-isopropanol (98∶1∶1) as the mobile phase. The flow rate was 0.5 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. A six-position 14-way valve and was equipped with 2 multi-center cutting valves was equipped to make multiple consecutive cuts of the pre-vitamin D₃ peak and vitamin D₃ peak. Results: The calibration curves showed a good linearity at the range of 1.018 4-5.092 mg·mL^-1(r≥0.999 8). The precision test showed the RSDs of the peak area of pre-vitamin D₃ and vitamin D₃ were 0.95% and 0.40%, respectively. The repeatability test showed the RSD of vitamin D₃ content was 0.41%. The average recovery rate (n=9) was 101.4%. The test solution was stable at 4 ℃ and 10 ℃ for 12 h, and the RSDs were 0.58% and 0.66%, respectively. The contents of vitamin D₃ in the samples of vitamin D drops measured by this method were 100.4%, 101.6%, 100.9%, 101.6%, 102.7% and 101.6%, which was basically consistent with the results measured by the fourth method in General Chapter 0722 of ChP 2020 Vol Ⅳ. Conclusion: This method has a good specificity and high sensitivity to accurately determine the content of vitamin D₃ in vitamin D drops.

Metabolomics, as a branch of systems biology, utilizes high-throughput omics technology to investigate metabolite changes within organisms, which enable us to explore the relationship between such alterations and disease etiology or evolution, thereby providing novel research insights for identifying relevant biomarkers and screening of diseases. In recent years, metabolomics has been widely used in the field of cancer research. At the same time, the changes of metabolic pathways can be deeply discussed by using the network shared database. Given that colorectal cancer ranks as the second most prevalent malignant tumor in China, it is crucial to search for clinically valuable tumor markers. This article will highlight the progress of recent five-year metabolomics studies in different matrices to identify potential biomarkers associated with colorectal cancer(CRC), so as to provide references for early screening of CRC.

Objective: To establish the RP-UPLC-PDA method for simultaneous determination of triptolide, triptonide, triptophenolide, wilforine, wilforlide A and celastrol in Tripterygii Radix. Methods: Tripterygii Radix were extracted with ethyl acetate and the extracts were dissolved and separated by methanol. The six components were determined by RP-UPLC-PDA method. The chromatographic column was Acquity^TM UPLC BEH C₁₈ column (100 mm×2.1 mm, 1.7 μm), the column temperature was 30 ℃, the injection volumn was 2 μL, the flow rate was 0.4 mL·min^-1 and the mobile phase was acetonitrile (A)-0.1% formic acid (B). Results: The linear relationship of six components was good (0.999 2≤r≤0.999 7) in the concentration ranges. The average recoveries were 99.2%-103.1% and RSDs were 1.2%-2.9%. The contents in 10 batches of Tripterygii Radix from different habitat were determined. The results showed that the contents of Tripterygii Radix in prepared pieces from different producing areas were different. The highest content of triptolide was 140.2 μg·g^-1, and the lowest content was 103.2 μg·g^-1. The highest content of triptonide was 224.7 μg·g^-1 and the lowest content was 112.2 μg·g^-1. The highest content of triptophenolide was 306.7 μg·g^-1 and the lowest content was 189.6 μg·g^-1. The highest and lowest contents of wilforine were 283.2 μg·g^-1 and 211.2 μg·g^-1. The highest content of wilfhabitat was 31.2 μg·g^-1 and the lowest content was 16.8 μg·g^-1. The highest content of celastrol was 87.6 μg·g^-1, and the lowest content was 52.1 μg·g^-1. Conclusion: The RP-UPLC-PDA method can simultaneously determine six components in Tripterygii Radix. The method is reliable and stable, which is suitable for quantitative analysis and determination of Tripterygii Radix.

Objective: To explore the biological function methods of mesenchymal stem cells(MSCs) for quality analysis. Methods: The surface markers of MSCs were detected by flow cytometry. MSCs osteogenic differentiation was induced by ascorbic acid and β-glycerophosphate sodium, etc., followed by Alizarin Red S staining. MSCs adipogenic differentiation was induced by IBMX, Rosiglitazone, etc., followed by Oil Red O staining. MSCs could differentiate into chondrocytes with treatment of ITS and TGFβ3, etc., followed by Alcian Blue staining. Cell co-culture of THP-1-macrophage with MSCs and ELISA assay were applied to detect the effects of MSCs on macrophage polarization. The expression levels of IL-10 and TNF α in the cell co-culture supernatant were detected by ELISA. To observe the effects of MSCs on lymphocyte proliferation, MSCs cultured with PBMCs, which were labeled with CFSE and activated by CD3/CD28, followed by flow cytometry. Results: The expression of MSCs surface markers, CD105, CD73, and CD90, was more than 95% respectively, while the expression of CD45, CD34, CD14, CD19, and HLA-DR expression was less than 2%. MSCs osteogenic differentiation assay showed red calcium nodules. Red lipid vacuoles were observed in MSCs adipogenic induction differentiation. Furthermore, MSCs have the differentiation potential to chondrocyte spheroids, and typical cartilage pits were observed. Co-culture of MSCs with THP-1 macrophages, an increase in IL-10 expression and downregulate TNFα secretion were observed. MSCs played inhibitory effects on the proliferation of PBMC activated by CD3/CD28, with an inhibition rate of 76.4%. Conclusions: This study established some of biological activity detection methods for MSCs, including MSCs surface markers, differentiation abilities, promotion of macrophage polarization, and inhibitory effects on lymphocyte proliferation. It provides a potential application for MSCs products quality control.

Objective: To establish a simple and efficient method for detecting telomerase activity for evaluating tumorigenic risk of cell products. Methods: Specific primers and probes were designed for the conserved domain of telomerase catalytic subunit (TERT), and the primers and probes of TERT gene were optimized and screened, and the primers and probes of internal reference genes were set up. Multiple quantitative PCR reaction was performed using two-color fluorescent probes in a reaction system, and RT-qPCR probe method was established. The expression of TERT gene in human mesenchymal stem cells HMSC was used to determine whether telomerase activity existed in the cells indirectly. Results: TERT gene of 293T/17 positive control cells could be stably and specifically detected by this method, with a mean Ct value of 23.96 and a RSD of 1.5. The internal reference gene GAPDH could be detected successfully, the mean Ct value was 14.13, the RSD was 1.3%. The reference gene GAPDH in MRC-5 could be detected in negative control cells, the mean Ct value was 12.81, the RSD was 0.46%, the TERT gene was not detected, and the telomerase activity was negative. The reference gene GAPDH in HMSC could be detected, the Ct mean value was 13.01, and the RSD was 3.8%, whereas, the telomerase activity of HMSC was negative. Conclusion: The real-time fluorescent quantitative RT-qPCR probe method established in this study can accurately detect the expression of catalytic subunit mRNA in telomerase positive cells with good repeatability and high specificity. It can be used to analyze telomerase activity of stem cells and indirectly evaluate tumorigenic risk of cell products derived from stem cells.

Objective: To study the dynamic changes of puerarin and icariin in plasma and hippocampus of mice, to observe the changes of depression-related protein expression in hippocampus, and to explore the pharmacokinetic changes of Shenge Bushen capsules (SBC) and its correlation with the changes of hippocampus-related protein expression. Methods: Puerarin and icariin in plasma and hippocampus of normal mice were analyzed by UPLC-MS/MS-ABSCIEX QTRAP 5500 triple quadrupole series linear ion trap mass spectrometry method using Waters Acquity HSS T3 (50 mm×2.1 mm, 1.8 μm) column with 0.05% formic acid water-acetonitrile-methanol (1∶1) containing 0.05% formic acid as mobile phases gradient elution at a flow rate of 0.2 mL·min^-1 and electrospray ion source under negative ion model. The proteins in hippocampus were expressed by Western blot assay. Results: Puerarin and icariin were detected in plasma and hippocampus after the oral administration, including plasma puerarin t_1/2 2.45 h, icariin t_1/2 3.59 h, hippocampus puerarin t_1/2 4.37 h and icariin t_1/2 8.5 h. The plasma concentration of puerarin accounts for 27.3% and that of icariin accounts for 1.34% of the dosage taken. Puerarin in hippocampus accounts for 2.47% of puerarin absorbed into blood, while icariin in hippocampus accounts for 73.56% of icariin absorbed into blood. The expressions of restrictive silencing factor (NRSF), brain-derived neurotrophic factor (BDNF) and its downstream protein tyrosine kinase B (TrkB), solute carrier transporter 6a4 (SLC6A4), glucocorticoid receptor (GR) and μ opioid receptor (MOR) in hippocampus were all up-regulated after SBC administration, in which icariin was negatively correlated with NRSF expression, puerarin was positively correlated with BDNF-TrkB and MOR, and GR had no obvious correlation with the proteins. Conclusions: Puerarin and icariin can be absorbed into the blood and distributed in the hippocampus after oral administration of SBC in mice. Icariin was easier to pass through the blood-brain barrier and distribute in the hippocampus than puerarin. The expression of hippocampus-related proteins was related to the changes of puerarin and icariin concentrations, suggesting that the targets of the two components as well as SBC were related to these proteins. This study provides an important experimental basis for the pharmacokinetic study of SBC and also provides a material basis of its antidepressant effect.

Objective: To establish a quantitative model to in-line in situ determination of the water content of domperidone tablets particles in fluidized bed granulation drying process. Methods: Using micro near infrared spectrometer, in-line collected near infrared(NIR) spectrum of domperidone tablets particles in granulation drying process, took sample at different drying time to acquire sample in different level of water content. The pretreatment method of first derivative(FD), multiplicative scatter correction (MSC) and Savitzky-Golay smoothing were selected. The modeling band was 1 100-1 600 nm, model was established by partial least square(PLS) method, and validation was executed including accuracy, precision, specificity, linearity and range, robustness. Results: The root mean square error of cross validation(RMSECV) was 0.079 7,the coefficient of determination(R²) was 0. 994 5,the root mean square error of prediction(RMSEP) was 0.092 3, the R² was 0.986 0. All validation results met predefined criteria. Conclusion: It is feasible for using micro NIR spectrometer to in-line measure the water content of domperidone tablets particles which provides an experimental basis for the in-line application of process analysis technology in pharmaceutical industry.

Objective: To conduct odor analysis of the main effective components, namely volatile oils, contained in two varieties, Notopterygium incisum Ting ex H.T.Chang and Notopterygium franchetii H.de Boiss, to provide a feasible method for promptly and accurately distinguishing between the differences in volatile oils of these two varieties of Notopterygii Rhizoma et Radix. This enriches the traditional evaluation content and serves as a reference for assessing the quality of extracts predominantly governed by volatile oils. Methods: The flavors of two samples of Notopterygii Rhizoma et Radix volatile oil were analyzed using electronic nose technology and sensory evaluation. The electronic nose data obtained were subjected to analysis and identification through principal component analysis (PCA) and linear discriminant analysis (LDA). Additionally, two nondestructive testing models-Fisher discrimination and multilayer perceptron (MLP) neural network discrimination were established for sample differentiation. Results: Sensory evaluation results indicated that pine resin flavor, cool flavor and woody flavor were the primary odor characteristics of both Notopterygii Rhizoma et Radix volatile oils. Additionally, the key flavor attribute influencing acceptance and differentiation was identified as spoiled yuba flavor, with the Notopterygium franchetii H. de Boiss volatile oil exhibiting a stronger presence of this attribute than the Notopterygium incisum Ting ex H. T. Chang volatile oil. The electronic nose results revealed that the nitrogen oxides’ response values in Notopterygium franchetii H. de Boiss volatile oil were significantly higher than those in Notopterygium incisum Ting ex H. T. Chang volatile oil. Meanwhile, the response values of hydrides, alcohol ether aldehydes, and ketones were slightly lower in Notopterygium franchetii H. de Boiss volatile oil compared to Notopterygium incisum Ting ex H. T. Chang volatile oil. The Fisher discriminant model demonstrated overall discrimination rates of 93.8% for the training set and 87.5% for the prediction set of the two volatile oils. In contrast, the MLP model achieved discrimination rates of 89.3% for the training set and 91.7% for the prediction set. Notably, the MLP model proved effective for identifying volatile oils, while the Fisher model exhibited greater suitability for discriminating volatile oils with broad-leaved characteristics. Conclusion: The combination of artificial senses and intelligent senses can be characterized from both subjective and objective perspectives, elucidating the flavor differences between the two kinds of Notopterygii Rhizoma et Radix volatile oils. The established Fisher discriminant function and MLP discriminant models can rapidly and accurately distinguish between the two kinds of Notopterygii Rhizoma et Radix volatiles. This lays a preliminary foundation for quality control in Notopterygii Rhizoma et Radix volatiles and offers new ideas and directions.

Objective: To establish an HPLC-principal component self-compare with correction factor method for quantification of two related substances (N-deacetyllappaconitine and ranaconitine) in lappaconitine hydrobromide injection. Methods: The analysis was performed on a Kromasil 300-5-C₁₈ (250 mm×4.6 mm,5 μm) column with a mobile phase composed of 0.04 mol·L^-1 potassium dihydrogen phosphate solution, methanol and acetonitrile (68∶17∶15). The detection wavelength was 252 nm, the flow rate was 0.8 mL·min^-1, the column temperature was 37 ℃, and the injection volume was 10 μL. The slope of linear equation was used to determine the relative correction factor between the two impurities and lappaconitine hydrobromide. The relative retention time was used to determine the position of related substances. The contents of two impurities in 25 batches of lappaconitine hydrobromide injection produced by four pharmaceutical companies were determined and compared with the results of the external standard method. Results: Lappaconitine hydrobromide and the impurities were separated well by this method. The relative retention time of N-deacetyllappaconitine and ranaconitine were 1.20 and 1.39, and the correction factors were 1.23 and 0.94, respectively. Lappaconitine hydrobromide, N-deacetyllappaconitine and ranaconitine showed good linearity in the mass concentration ranges of 0.951 7-38.07 μg·mL^-1, 1.047-41.87 μg·mL^-1 and 1.001-40.02 μg·mL^-1 with r=1.000, respectively. The average recovery rates of lappaconitine hydrobromide, N-deacetyllappaconitine and ranaconitine were 100.2%, 100.5% and 100.5% respectively, with RSD less than 2.0%. The limits of detection (LOD) of lappaconitine hydrobromide, N-deacetyllappaconitine and ranaconitine were 0.095, 0.10, 0.10 μg·mL^-1 and the limits of quantitation (LOQ) were 0.32, 0.35 and 0.33 μg·mL^-1, respectively. The content of N-deacetyllappaconitine in 25 batches of lappaconitine hydrobromide injection was in the range of 0.31%-0.82% and the content of ranaconitine was in the range of 0%-0.09%. It was consistent with the determination result of the external standard method. Conclusion: The method is proved to be simple, rapid, and accurate for the determination of related substances in lappaconitine hydrobromide injection.

Objective: To analyze and optimize stir-baked with vinegar technology of Genkwa Flos decoction pieces by multi-index weighted scoring method based on orthogonal experiment, and to provide technical index on standardization of stir-baked with vinegar technology of Genkwa Flos decoction pieces. Methods: The orthogonal experiment was employed to analyze and evaluate the four key factors related to stir-baked with vinegar technology including the amount of vinegar, dampening time, temperature and stir-baked time by the six main indicators including the total contents of chlorogenic acid, tiliroside, luteolin, apigenin, hydroxygenkwanin and genkwanin, the content of alcohol extractive and appearance characters. Results: The amount of vinegar and stir-baked temperature had notable effects, while the dampening time and stir-baked time had no notable effects on the test results. Considering comprehensively the effects of four key factors, the optimized stir-baked with vinegar technology was as follows: add 0.3 kg of vinegar to 1 kg of Genkwa Flos, and fry at 160 ℃ for 10 min after dampening for 15 min. Conclusion: This study offers guidance and reference for standardizing the stir-baked with vinegar technology of Genkwa Flos decoction pieces.

Objective: To establish an HPLC method for simultaneous determination of loganic acid, gentiopicroside, hesperidin, paeonol, aloe-emodin, rhein, emodin, chrysophanol and physcion in Tongshu Koushuang tablets (Rhei Radix et Rhizoma, Aurantii Fructus Immaturus and Gentianae Macrophyllae Radix, etc.). Methods: The samples were extracted by refluxing with methanol and the separation was performed on a Thermo Acclaim-C₁₈(250 mm×4.6 mm, 5 μm) column at 30 ℃, with mobile phase consisting of 0.15% phosphoric acid solution and acetonitrile with a gradient elution at a flow rate of 1.0 mL·min^-1. The injection volume was 10 μL, and the detection wavelength was set at 236 nm (detecting loganic acid), 280 nm (detecting gentiopicroside, hesperidin and paeonol) and 254 nm (detecting aloe-emodin, rhein, emodin, chrysophanol and physcion). Results: Nine constituents (loganic acid, gentiopicroside, hesperidin, paeonol, aloe-emodin, rhein, emodin, chrysophanol and physcion) were in good linearity within their own ranges (r were 0.999 1-0.999 9), and the average recoveries were 99.6% (RSD=1.9%), 101.1% (RSD=2.0%), 103.0% (RSD=1.5%), 101.6% (RSD=0.51%), 99.8% (RSD=1.4%), 100.9% (RSD=1.4%), 100.8% (RSD=1.8%), 102.7% (RSD=1.2%) and 102.9% (RSD=0.86%), respectively. The content ranges of nine constituents were 1.871-3.517 mg·g^-1, 6.274-12.55 mg·g^-1, 2.140-3.946 mg·g^-1, 1.393-2.018 mg·g^-1, 0.394 8-0.807 2 mg·g^-1, 0.657 5-1.246 mg·g^-1, 0.484 7-0.915 8 mg·g^-1, 0.839 8-1.429 mg·g^-1 and 0.327 3-0.547 9 mg·g^-1, respectively. Conclusion: This simple and accurate method can be used for the quality control and evaluation of Tongshu Koushuang tablets.

Objective: To investigate the source and quality of commercially available Epimedium based on HPLC fingerprint and multi-index assay, so as to provide reference for its quality evaluation and resource development. Methods: Forty batches of commercially available epimedium were collected. Fingerprint analysis and determination of the total flavonoid glycosides were performed by HPLC method. The total flavonoid glycosides were determined by UV method. And the contents of their extracts were determined. Stoichiometric analysis was employed to perform the comprehensive quality evaluation of different medicinal materials of Epimedium. Results: There were 9 different plant sources of Epimedium in the market. The five authentic Epimedium were of good quality, and the local varieties were coarse Epimedium. The contents were high in Epimedium from Hunan, while the contents were low in Epimedium from Qianling and Sichuan. The comprehensive quality ranking of Epimedium indicated that Epimedium Fletchum and Epimedium Fletchum ranked first by the principal component analysis method. The ranking of the same variety varied greatly with different habitats, and the Qianling Epimedium and adulterated Epimedium ranked last. The results of cluster analysis and principal component analysis were basically consistent. The cluster analysis showed distinguishing significance in plant sources, habitats, processing methods and contents. Based on the common pattern of E. brevicornu, a total of 8 common peaks were identified for 9 species of E. Brevicornu, and their similarities ranged from 0.066 to 0.979. Differences were observed among different species. Conclusion: Both the fingerprint similarity and the chemical model of content determination can be used to evaluate the species and quality of Epimedium.

Nuclear magnetic resonance technology is not only an excellent qualitative technique, but also has its unique advantages in quantitative aspects. As a highly selective, fast, and economical quantitative analysis method, nuclear magnetic quantification technology has been widely applied in fields such as medicine, chemical industry, and food. The basic principle of nuclear magnetic resonance quantitative technology is that the strength of the detected magnetic resonance signal is proportional to the number of corresponding nuclear. Based on the properties of the sample, corresponding technical means and quantitative methods can be flexibly selected, which have wide application value. This article provides a systematic summary of the quantitative methods of nuclear magnetic quantification technology combined with relevant literature reports and research results, providing reference for its practical application and research.

Objective: To establish a laser diffraction method for the determination of particle size distribution of budesonide suspension for inhalation. Methods: The Master sizer 3000 laser size analyzer with the Hydro SV wet sampler was used. The sample solution should be immediately measured after ultrasonication. The measurement conditions were as follows:refractive index of budesonide was 1.592, absorbency of budesonide was 0.01, dispersing medium was saturated solution of budesonide containing 0.5% Tween 80 with refractive index of 1.33, stirring rate was 1 000 r·min^-1,and laser obscurations were 6%-12%. Results: RSD of D₅₀ was lower than 2%, RSDs of D₁₀ and D₉₀ were both lower than 5%.The particle size distribution D₉₀ values of 3 batches of budesonide suspension for inhalation were less than 5 μm. Conclusion: The method is simple, accurate and producible, which is suitable for the particle size detection of budesonide suspension for inhalation.

Objective: To investigate the feasibility of applying for bioequivalence exemption based on parallel artificial membrane permeability assay (PAMPA) data by evaluating the solubility and in vitro permeability of levofloxacin, comparing the prescription differences between the generic and reference formulations, assessing the impact of disparate excipients on the permeation behavior of the active pharmaceutical ingredient (API), and predicting the bioequivalence of the two formulations. Methods: The dissolution of the raw material at pH 1.0 to pH 6.8 was determined using high-performance liquid chromatography. The μFlux^TM system was employed to determine the permeability of the raw material and the mixture of raw material with sodium stearyl fumarate in various media. pH 5.0 fed-state intestinal fluid, pH 6.5 fasted-state intestinal fluid, and pH 7.4 phosphate buffer (16 mL) were precisely added to the donor chamber, while 16 mL of Accepter Sink Buffer was added to the receptor chamber. The rotor speed was set at 200 r·min^-1, and the collection time was 180 min. Determine the permeability of the raw material and the mixture of raw material with sodium stearyl fumarate in various media, as well as the permeability of the raw material in the reference and generic formulations powders. The impact of newly added excipients and other changed excipients on API was evaluated through a two-tailed t-test. The Macro Flux^TM system was used to measure the dissolution-permeation curves of the reference and generic formulations, intestinal simulation fluids at pH 5.0 and pH 6.5 (1 000 mL) were added to the dissolution cup as dissolution media, with a stirring speed of 75 r·min^-1 using a paddle method, 12 mL of Accepter Sink Buffer was added to the receptor chamber, and the stirring speed of the micro-stirring rod was set at 450 r·min^-1, one tablet of each reference or generic formulation was placed in the dissolution cup, and the dissolution-permeation curves of the formulations were measured. The similarity of dissolution curves was compared, and the permeability rate (J_Flux) and cumulative drug permeation amount (AMT) were calculated to predict the bioequivalence of the two formulations, ensuring that the 90% confidence interval for the geometric mean ratio of J_Flux and AMT of the two formulations fell within the range of 80% to 125%. Results: Levofloxacin solubility ranged from 16.4 mg·mL^-1 to 62.7 mg·mL^-1 across different mediums, its permeability in pH 5.0 fed-state simulated intestinal fluid, pH 6.5 fasted-state simulated intestinal fluid, and pH 7.4 phosphate-buffered saline was 2.92×10^-6 cm·s^-1, 1.01×10^-5 cm·s^-1, and 1.07×10^-5 cm·s^-1, respectively. The addition of sodium stearyl fumarate showed no significant difference in permeability compared to the original API (P＜0.05), and there were no significant differences between the powder API of the reference and generic formulations (P＜0.05). The dissolution curves of both formulations were similar, with the 90% confidence interval for JFlux and AMT within the predefined range. Conclusion: Levofloxacin tablets, classified as BCS Class Ⅰ, demonstrated that the altered excipients in the formulation did not impact on the API's permeability, confirming bioequivalence between the reference and generic formulations. The bioequivalence exemption study based on PAMPA can be utilized for permeability studies of raw materials, excipient screening and optimization, and prediction of formulation bioequivalence, effectively reducing drug development costs and time. This study provides reference data for pharmaceutical companies applying for bioequivalence exemptions.

Objective: To establish a multi-component content determination and characteristic chromatogram method for Jieyu Anshen granules, combined with chemometrics, to comprehensively evaluate the quality of Jieyu Anshen granules. Methods: The separation was performed on a CAPCELL PAK MG Ⅱ C₁₈(250 mm×4.6 mm, 5 μm) column using acetonitrile (A) and 0.1% phosphoric acid aqueous solution (B) as the mobile phases with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃ and the detection wavelengths were set at 237 nm and 335 nm. The analysis method was validated. Based on the multi-component content determination method, characteristic chromatogram was established. The samples were analyzed and evaluated through chemometrics to identify the main factors affecting their quality. Results: A multi-component content determination method was established for Jieyu Anshen granules. The five components had good linear relationships within their respective ranges, with recovery rates ranging from 95.6% to 100.0%. And 145 batches of samples were tested, and a total of 54 batches were found to be unqualified, with a total unqualified rate of 37.2%. Establishing a characteristic chromatogram of Anshen granules, 12 characteristic peaks were attributed to 6 medicinal herbs. Through chemometric analysis, it was found that there were certain differences in the quality of samples from 10 manufacturers. The quality of fried gardenia and stir-baked licorice was the key factor. Conclusion: The established multi-component content determination method and characteristic chromatogram method for Jieyu Anshen granules are convenient and reliable, and can be used for comprehensive evaluation of the quality of Jieyu Anshen granules.

Objective: To establish a method for identifying Fritillariae Cirrhosae Bulbus and its adulterant Fritillariae Ussuriensis Bulbus. Methods: Chemometrics techniques were used for the analysis of the sample data from determination with Q TOF MS, and characteristic ion pairs were selected as m/z 578.3→164.14 and m/z 578.3→398.31 with triple quadrupole mass spectrometry applied, respectively. Then a specific approach was developed which used Waters Acquisition UPLC CSH (75 mm×2.1 mm, 1.7 μm) as column, and with acetonitrile and 0.1% formic acid solution as the mobile phase, and a flow rate of 0.4 mL·min^-1. Two ion pairs: m/z 578.3→164.14 and m/z 578.3→398.31 were detected by triple quadrupole mass spectrometer with MRM mode. Results: The validation results through 108 batches of samples indicated that the two characteristic ion pairs, m/z 578.3→164.14 and m/z 578.3→398.31, could effectively distinguish between Fritillariae Cirrhosae Bulbus and its adulterant Fritillariae Ussuriensis Bulbus. Conclusion: The method had good specificity and sensitivity, and could be used for the detection of adulterated Fritillariae Cirrhosae Bulbus with Fritillariae Ussuriensis Bulbus.

Objective: To establish an UHPLC-MS/MS method for determining eight primary components (catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin) in the young branches and leaves of Xanthoceras sorbifolia Bunge, a medicinal plant from Mongolia, and to compare their contents in samples at different growth stages. Methods: A Waters CORTECS C₁₈ (100 mm×2.1 mm, 1.6 μm) chromatographic column was adopted using the mobile phase comprised of water containing 0.1% formic acid (A) and acetonitrile (B) with gradient elution(0-1 min, 5%B; 1-10 min, 5%B→28%B; 10-11 min, 28%B→95%B; 11-14 min, 95%B; 14-15 min, 95%B→5%B) at a flow rate of 0.3 mL·min^-1. The temperature of the column was set at 40 ℃. Injecting volume was 2 μL. Detection was conducted using electrospray ionization (ESI) in negative ion mode with multiple reaction monitoring (MRM). Results: The linearity of the eight chemical components was found to be excellent in the tested concentration ranges, with correlation coefficients above 0.997 6. Precision, repeatability and stability were satisfactory and the average recoveries were between 97.4% and 106.0% with RSDs≤5.0%. In six batches of leaves, contents of catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin were in the ranges of 0.090-0.904 mg·g^-1, 0.093-2.258 mg·g^-1, 0.001-0.005 mg·g^-1, 0.530-6.176 mg·g^-1, 0.158-1.561 mg·g^-1, 0.002-0.056 mg·g^-1, 4.008-10.218 mg·g^-1 and 1.049-16.990 mg·g^-1, respectively. In six batches of young branches, the contents ranged from 0.384-1.025 mg·g^-1, 0.911-2.427 mg·g^-1, 0.008-0.127 mg·g^-1, 0.870-2.295 mg·g^-1, 0.659-1.746 mg·g^-1, 0.125-1.079 mg·g^-1, 2.296-4.681 mg·g^-1 and 1.958-4.946 mg·g^-1, respectively. The contents of eight components varied a lot in samples from different parts. The contents of myricitrin, rutin and quercitrin in the leaves exhibited noticeable changes with the growth cycle, suggesting their potential as quality control markers for leaves of Xanthoceras sorbifolia. Conclusion: The method is accurate, sensitive, stable, repeatable, and suitable for simultaneous determination of eight components in Xanthoceras sorbifolia Bunge, offering reference for quality control of its leaves and branches.