To establish a method for identifying Fritillariae Cirrhosae Bulbus and its adulterant Fritillariae Ussuriensis Bulbus.

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.

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.

The method had good specificity and sensitivity, and could be used for the detection of adulterated Fritillariae Cirrhosae Bulbus with Fritillariae Ussuriensis Bulbus.

To establish a fingerprint of Gualou Guizhi decoction and a method for determination of multi-component to clarify the transfer rule of quantities and quality from decoction pieces and substance benchmarks.

Fifteen batches of Gualou Guizhi decoction substance benchmarks were prepared and analyzed by the method of HPLC. Traditional Chinese medicine (TCM) Chromatographic Fingerprint Similarity Evaluation Software (2012) and hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to evaluate the quality differences between batches of substance benchmarks, and screen the main chemical compositions that were led to quality discrepancy. The main differential components were determined by HPLC. And their contents, yields and transfer rates were analyzed for transfer rule of quantities and quality.

HPLC fingerprint of Gualou Guizhi decoction was established. A total of 30 common peaks in the fingerprint of Gualou Guizhi decoction substance benchmarks were assigned. Compared with the reference standards, 9 components were identified as gallic acid, albiflorin, paeoniflorin, liquiritin, ononin, liquiritigenin, cinnamic acid, isoliquiritigenin and glycyrrhizic acid. The similarities of 15 batches of Gualou Guizhi decoction substance benchmarks samples were all above 0.920. All of them were divided into two categories by three chemical recognition modes for 5 major differential components, including glycyrrhizic acid, liquiritigenin, paeoniflorin, iquiritin, and ononin. The HPLC was also applied to determine the contents of multi-components and the method validation results were good. The average transfer rates of five index components were 51.65%, 40.46%, 74.74%, 60.06%, and 34.54%, respectively and their average extraction rate was 14.20%.

The critical quality properties of Gualou Guizhi decoction can be stably transferred from decoction pieces to substance benchmarks. The method of fingerprint and content determination is accurate and reliable, which is able to provide technique for quality control of Gualou Guizhi decoction formulation.

To analyze the structure of the maximum impurity in halometasone/triclosan cream and discuss the rationality of the current registered standard for imported drugs.

The related substances in halometasone/triclosan were detected according to JX20080304, and dioxins were determined according to USP. Finally, the possible structures of the maximum impurity were characterized by LC-Q TOF/MS.

The results showed that the maximum unknown impurity(RRT 0.67), calculated according to halometasone was OOS(1.0%) at 2.5%. The peak was related to triclosan through comparing with the chromatography behavior of triclosan in reference substance solution. MS information indicted the process impurities of triclosan.

For compound preparations, to ensure the stability and safety of the product, the impurities relevant to different components should be controlled separately at rational limit based on safety data.

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.

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%.

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.

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.

To establish an HPLC-MS/MS method for the simultaneous determination of six characteristic components of Scutellariae Radix including baicalein, baicalin, wogonin, wogonoside, chrysin and oroxylin A in rat plasma, and to compare their pharmacokinetic profiles after oral administration of Scutellariae Radix and Niuhuang Jiedu tablets (NHJDT) to rats.

Rats were given 250 mg·kg^-1 of NHJDT suspension or the prescribed equivalent amount of Scutellariae Radix aqueous extract, and plasma samples were collected at different time intervals. Naringenin was used as internal standard. After precipitated with methanol, the plasma samples were separated on an Inertsil C₈-3 (150 mm×4.6 mm, 5 μm) column by linear gradient elution with 0.1% formic acid solution(A)-0.1% formic acid-methanol(B) as mobile phase. The flow rate was 1.0 mL·min^-1, and the column temperature was 35 ℃. The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source combined with multiple reaction monitoring(MRM) mode in positive ion mode. The pharmacokinetic parameters were calculated and statistically analyzed.

The six characteristic components of Scutellariae Radix were all in good linear relationships in the range of 5-500 ng·mL^-1. The RSDs for accuracy test were in the range of 88.43%-108.6%, and the RSDs for inter-batch and intra-batch precision tests were all below 15%. The matrix effect and plasma stability met the requirements of methodology validation in biological sample analysis. Baicalin and wogonoside were the major components detected in rat plasma after oral gavage of Scutellariae Radix aqueous extract and NHJDT suspension. The AUC_0-t of wogonoside was significantly increased in NHJDT group compared with Scutellariae Radix group. Furthermore, the C_max of wogonoside and baicalin were significantly increased while the T_max was decreased after NHJDT suspension administration.

This method is specific and sensitive for the determination of six characteristic components of Scutellariae Radix, and suitable for pharmacokinetic study of rat plasma. NHJDT with co-existing components enhances the absorption and influences the pharmacokinetic behaviors of active ingredients of Scutellariae Radix.

To establish an effective method for detecting the components of Lychas mucronatus and investigating instances of adulteration with whole scorpions in Chinese patent medicines.

Thirty Chinese patent medicines containing whole scorpion components, as documented in the Pharmacopoeia of the People’s Republic of China, were collected. DNA was extracted, and differences in the mitochondrial cytochrome C oxidase subunit Ⅰ(COⅠ) gene sequences were compared. Specific primers for the identification of Mesobuthus martensii and Lychas mucronatus DNA were designed. Annealing temperature, cycle number, Taq polymerase, and DNA concentration were optimized to establish the most suitable PCR identification system and conditions. The optimal PCR conditions for distinguishing the sharp-tailed wolf scorpion were finally determined to be an annealing temperature of 52 ℃, 36 cycles, 2×PCR Mix Taq enzyme, and 1 μL of DNA template. The developed method was applied to identify the presence of Centruroides gracilis components in 30 commercially available Chinese patent medicines.

Gel electrophoresis revealed the presence of authentic DNA bands specific to the Mesobuthus martensii in all 30 whole scorpion-containing Chinese patent medicines. Eighteen medicines (60%) showed a single specific DNA band between 100-250 bp, indicative of Lychas mucronatus, while the blank control exhibited no bands. Sequencing of the PCR products containing the identified Lychas mucronatus -specific bands, after TA cloning, revealed sequences with high similarity (92%-98%) to the reference sequences of Lychas mucronatus. Phylogenetic analysis indicated that the obtained sequences belonged to the same lineage as Lychas mucronatus and were distinct from the Mesobuthus martensii. Therefore, the presence of a single band around 120 bp indicates the identification of Lychas mucronatus.

The presence of Lychas mucronatus components mixed with authentic scorpion materials in Chinese patent medicines indicates a need for strengthened supervision. The method established in this study enables the rapid and accurate detection of Lychas mucronatus components in Chinese patent medicines. This is beneficial for improving the quality control of whole scorpion-containing Chinese patent medicines, providing assurance for their clinical applications.

To establish a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of nine effective components including phenolic acids and flavonoids (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutoside, isoquercitrin, kaemperol-3-O-rutinoside, 3，4-O-dicaffeoylquinic acid, 3，5-O-dicaffeoylquinic acid, 4，5-O-dicaffeoylquinic acid), in Solidaginis Herba.

Ultra-high performance liquid chromatography was developed on an ACQUITY UPLC^® HSS T3 (100 mm×2.1 mm, 1.8 μm) column with acetonitrile (A)-0.1% phosphoric acid solution (B) as mobile phase in gradient elution. The flow rate was 0.3 mL·min^-1, the detection wavelength was 327 nm, the column temperature was 30 ℃, and the injection volume was 1 μL. Chlorogenic acid and rutoside were chosen as the internal standards for phenolic acids and flavonoids respectively to establish the determine correction factors of neochlorogenic acid, cryptochlorogenic acid，3，4-O-dicaffeoylquinic acid, 3，5-O-dicaffeoylquinic acid, 4，5-O-dicaffeoylquinic acid isoquercitrin and kaemperol-3-O-rutinoside, respectively, by multi-point correction method and their contents were calculated. The results were compared with the results of external standard method to verify their accuracy.

Nine components showed good linear relationships within their own ranges(r≥0.999 5), whose average recoveries were 98.2%-101.7% with the RSDs of 1.0%-1.7%. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutoside, isoquercitrin, kaemperol-3-O-rutinoside, 3，4-O-dicaffeoylquinic acid, 3，5-O-dicaffeoylquinic acid and 4，5-O-dicaffeoylquinic acid, in nine batches of samples were 0.008%-0.014%, 0.055%-0.097%, 0.010%-0.019%, 0.191%-0.412%, 0.018%-0.035%, 0.116%-0.50%, 0.028%-0.048%, 0.111%-0.235% and 0.113%-0.182%. The results obtained by QAMS were consistent with those of external standard method(relative average deviations less than 2.8%).

The simple, accurate and reproducible method can be used for the quality control of Solidaginis Herba.

To explore the method of pharmacognosic identification of four medicinal seeds, such as Plantaginis Semen from two kinds of original plant，Allii Fistulosi Semen, Allii Tuberosi Semen, and to digitize the identification characteristics.

The seeds of the four medicinal plants were studied by means of type microscope and light microscope, including the appearance(positive view, bottom view, hilum view and surface feature), internal morphology(cross profile and longitudinal profile) and the characters of micromorphology(transverse section, powder). The characteristics of the four medicinal seeds were summarized, and the identification points were characterized by digital method.

The seeds of the four medicinal plants could be identified according to the appearance, internal morphology and the characters of micromorphology.

This study lays a foundation for the variety identification and provides reference for the standard formulation and quality control of the four medicinal seeds. It also provides ideas for digital characterization of seed medicinal materials.

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.

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.

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.

This method has a good specificity and high sensitivity to accurately determine the content of vitamin D₃ in vitamin D drops.

To establish high-performance liquid chromatography method using cyclodextrin as a mobile phase additive for the determination of ursolic acid and oleanolic acid in Corni Fructus, providing a basis for quality control of Corni Fructus.

Molecular design and response surface method were used to optimize the determination method of ursolic acid and oleanolic acid in Corni Fructus. Agilent Eclipse XDB C₁₈ column (150 mm×4.6 mm, 5 μm) was adopted as the stationary phase, the mobile phase was 3 mmol·L^-1 γ-cyclodextrin solution (contained 0.05% phosphoric acid)-acetonitrile (60∶40), the flow rate was 1.0 mL·min^-1, the column temperature was 25 ℃ and the detecting wavelength was 210 nm.

The resolution of chromatographic peaks of ursolic acid and oleanolic acid was 3.81. Linearities were good in the range of 400-4 000 ng for ursolic acid and in the range of 200-2 000 ng for oleanolic acid. The RSDs for precision, reproducibility, and stability were less than 1.4%, 1.8%, and 1.7%, respectively. The recovery rates were 98.5% (RSD=1.7%) and 99.1% (RSD=2.0%). The contents of ursolic acid in three batches of Corni Fructus were 2.66 mg·g^-1, 2.35 mg·g^-1 and 2.91 mg·g^-1, and those of oleanolic acid were 0.83 mg·g^-1, 0.78 mg·g^-1and 1.12 mg·g^-1, respectively.

This method is simple, accurate, reliable, and environmental friendly, and can be used as a quality control for Corni Fructus.