To establish a quantitative analysis of multi-components by single marker (QAMS) method to simultaneously determine the contents of kalangin, diphenylheptane A, galangin, kaempferin and galangin-3-O-methyl ether.

Hypersil ODS2 C₁₈ (250 mm×4.6 mm, 5 μm) was used by high performance liquid chromatography (HPLC) and was eluted with 0.3% acetic acid in water (A) and acetonitrile (B) (0-2 min, 10%B→15%B；2-5 min, 15%B→20%B；5-20 min, 20%B→35%B；20-30 min, 35%B→40%B；30-55 min, 40%B；55-60 min, 40%B→100%B), the column temperature was 30 ℃, the volume flow rate was 0.8 mL·min^-1, and the detection wavelength was 254 nm. The relative correction factors of each chemical component were calculated with gingerin as the internal reference, and the content was determined, which was compared with the results of the external standard method, and the quality of galangal from different origins was evaluated by chemical pattern recognition.

Kaempferol, diphenylheptane A, galangin, kaempferin and galangin-3-O-methyl ether had a good linear relationship in the ranges of 0.005 5-0.110 0, 0.140 0-2.800 0, 0.149 6-2.992 0, 0.021 5-0.430 0 and 0.022 2-0.444 0 μg, respectively, and the average sample recoveries were 100.7%, 101.4%, 99.9%, 100.9% and 101.7%, respectively. The RSDs were 1.3%, 2.8%, 0.83%, 1.4% and 1.8%, respectively. Quantitative analysis of multicomponents by single marker method and external standard method was no significant difference in the results.

This method is rapid, accurate and specific, which can provide a reference for the quality control of galangal.

To determine the contents of 28 elements in Xianzhuli under processes of dry distillation and fire preparation by inductively coupled plasma mass spectrometry (ICP-MS), and evaluate the rationality of the current temperature used in the dry distillation process as well as whether the preparation process of commercial samples conforms to the traditional or modern one.

The samples of Xianzhuli were pre-treated with nitric acid before microwave digestion. The contents of 28 elements in 59 batches of Xianzhuli samples were determined by inductively coupled plasma mass spectrometry(ICP-MS), and the methodology was investigated.

The standard curve of 28 elements had a good linear relationship with r≥0.999 2. The detection limits were 0.007 1-1.249 5 ng·mL^-1, the RSDs for precision, repeatability and repeatability tests were 0.30%-3.5%, 0.69%-6.4% and 1.1%-3.3%, respectively and the recovery rates were 88.6%-105.5% with RSDs ranged from 1.0% to 3.2%. The contents of Na, Mg, K, Ca, Mn, Fe, Zn and Rb in 59 batches of Xianzhuli were high, and the contents of heavy metal elements did not exceed the limit requirements. The results of cluster analysis showed that the samples prepared by dry distillation and fire preparation were clustered into one class. By principal component analysis, 7 principal components were obtained, and the cumulative variance rate was 75.6%. Mg, K, Ca, Cr, Fe, Co, Cu, Rb, Cd, Ba, Tl and Pb were identified as the characteristic elements of Xianzhuli.

Based on the contents of beneficial and harmful elements, it is found that the contents of toxic and harmful elements in the sample processed with 160 ℃ dry distillation are lower than processed with fire preparation. And the contents of beneficial elements are similar in two kinds of samples. This temperature is consistent with the dry distillation process temperature approved by provincial drug regulatory authorities for currently certified enterprises. Most of the commercially available samples meet the requirements of traditional or modern processes, and a few may have quality differences due to different processes. The determination and analysis of heavy metals and other elements in Xianzhuli under different processes can provided scientific basis for safe production and rational use of Xianzhuli.

To establish a method for the simultaneous determination of six components (rutin, naringin, neohesperidin, quercetin, purpurin and mollugin) in Huazhi tablets and Huazhi capsules by HPLC.

The assay was performed on an Agilent ZORBAX SB-Aq (150 mm×4.6 mm, 5 μm) and the sample was eluted by mobile phase consisting of acetonitrile(A) -0.1% phosphoric acid(B) with a gradient at a flow rate of 1.0 mL·min^-1 and the column temperature was 30 ℃. The detection wavelengths were set at 250 nm for rutin during 0-20 min, 283 nm for naringin and neohesperidin during 20-37 min, and 250 nm for quercetin, purpurin and mollugin during 37-60 min.

Rutin, naringin, neohesperidin, quercetin, purpurin and mollugin exhibited good linearity(r>0.999 0) in the ranges of 37.26-1 863.20 μg·mL^-1, 11.27-563.70 μg·mL^-1, 9.58-479.04 μg·mL^-1, 1.92-95.90 μg·mL^-1, 0.52-25.83 μg·mL^-1 and 0.90-45.10 μg·mL^-1, respectively. The average recoveries of the above mentioned six components in Huazhi tablets (n=6) were 103.2%(RSD=1.4%), 99.5%(RSD=1.7%), 97.7%(RSD=1.2%), 95.2%(RSD=1.1%), 104.2% (RSD=1.2%) and 104.2%(RSD=0.80%), respectively, and the average recoveries of the above mentioned six components in Huazhi capsules (n=6) were 102.5% (RSD=1.3%), 96.9%(RSD=0.48%), 97.1%(RSD=1.1%), 96.9%(RSD=0.78%), 102.3%(RSD=1.4%) and 101.8%(RSD=1.2%), respectively. The content ranges (mean ± SD) of rutin, naringin, neohesperidin, quercetin, purpurin and mollugin in the 17 batches of Huazhi tablets and Huazhi capsules were 31.14-98.25(44.33±15.65) mg·g^-1, 5.12-20.85(12.96±5.85) mg·g^-1, 4.03-17.00(10.14±5.17) mg·g^-1, 0.63-7.17(1.97±1.49) mg·g^-1, 0.23-1.32(0.57±0.28) mg·g^-1 and 0.67-1.72(1.08±0.29) mg·g^-1, respectively.

The established method can simultaneously determine six components in Huazhi tablets and Huazhi capsules and can provide a reference for the quality control of Huazhi tablets and Huazhi capsules.

To investigate different batches of Runbi Tongqiao drops by fingerprint and multi-index component content determination method, and to provide basis for its quality evaluation.

Acclaim TM-C₁₈ chromatographic column(250 mm×4.6 mm, 5 μm) was used with methanol(A)-acetonitrile(B) -0.2% formic acid aqueous solution(C) as the mobile phase with gradient elution. The flow rate was 0.8-1.0 mL·min^-1. The injection volume was 10 μL and the detection wavelength was 327 nm. The HPLC fingerprint of Runbi Tongqiao drops was established, and the quality consistency of Runbi Tongqiao drops was compared by stoichiometric method. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, luteoloside and isochlorogenic acid C were determined by HPLC.

A total of 14 common peaks were calibrated in the fingerprints of 10 batches of samples, and the similarities were ≥0.933. Seven common peaks of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, luteoloside and isochlorogenic acid C were identified by reference substances. The contents of seven components in Runbi Tongqiao drops were determined simultaneously, which were 0.131 5-0.133 8 mg·mL^-1, 0.095 5-0.098 9 mg·mL^-1, 0.087 4-0.090 1 mg·mL^-1, 0.012 8-0.015 8 mg·mL^-1, 0.010 3-0.013 7 mg·mL^-1, 0.022 7-0.024 9 mg·mL^-1 and 0.006 8-0.008 6 mg·mL^-1, respectively.

The established fingerprint of Runbi Tongqiao drops is stable and reliable, and the simultaneous determination method of multi-component content is simple and fast. It can be used for the quality control of Runbi Tongqiao drops, which lays a solid foundation for the follow-up study of Runbi Tongqiao drops.

To develop a rapid, specific and sensitive UPLC-MS/MS method for the determination of vonoprazan in human plasma and its application in a bioequivalence study of two types of tablets.

A single dose, two-cycle, two products, and self-cross controlled trial design on bioequivalence was used. Plasma samples were collected from healthy human volunteers at different time points after oral administration with the test or reference product of 20 mg fumarate vonoprazan tablets under both fasting and fed conditions, respectively. The plasma samples were treated by acetonitrile protein precipitation and then analyzed by UPLC-MS/MS. Chromatographic separation of vonoprazan was achieved using a Waters ACQUITY UPLC® BEH C₁₈（50 mm × 2.1 mm, 1.7 μm）column at 40 ℃. The mobile phase consisted of water (containing 0.1% formic acid) for eluent A and acetonitrile (containing 0.1% formic acid) for eluent B under a gradient elution. An electrospray ionization (ESI) with multiple reaction monitoring (MRM) mode was used to monitor the precursor-product ion transitions of m/z 346.1→315.4 for vonoprazan and m/z 350.1→316.2 for vonoprazan-d₄.

The rang of linearity was 0.30-50.00 ng·mL^-1（r>0.998 9）, and the LLOQ was 0.30 ng·mL^-1. Intra- and inter-day precision values were within 5.7%, and intra- and inter-day accuracy values were ranged from -2.15% to 0.82%. Recovery, specificity, matrix effect and stability met the guiding principles. This method has been successfully applied to study the bioequivalence of vonoprazan fumarate tablets. The C_max of the test product in postprandial and fasting tests were (29.08±11.59) ng·mL^-1 and (26.87±8.14) ng·mL^-1, respectively, and the AUC_0-t was (258.90±87.71) h·ng·mL^-1 and (223.08±43.27) h·ng·mL^-1, respectively. The C_max of the reference product in postprandial and fasting tests were (28.73±10.25) ng·mL^-1 and (26.93±8.09) ng·mL^-1, respectively, and the AUC_0-t was (250.33±73.13) h·ng·mL^-1 and (227.56±46.26) h·ng·mL^-1, respectively. In the postprandial trial, the 90% CIs for the geometric mean ratios of C_max, AUC_0-t and AUC_0-∞ of the test and reference products were 88.64%-112.28%, 96.1%-108.2% and 96.6%-108.7%, respectively. And in the fasting trail, the 90% CIs for the geometric mean ratios of C_max, AUC_0-t and AUC_0-∞ of the test and reference products were 94.01%-106.23%, 94.71%-102.03% and 95.18%-102.47%, respectively.

This validated method has the advantages of simplicity, rapid, and high sensitivity. Test vonoprazan fumarate tablets are bioequivalent to the reference product.

To establish the HPLC fingerprint and multi-component quantitative analysis of Hanchuang Zupa granules, and evaluate the quality of multiple batches of Hanchuang Zupa granules by chemical pattern recognition technology.

The sample pretreatment conditions and chromatographic analysis conditions of Hanchuang Zupa granules were optimized, and the optimal HPLC fingerprint and multi-component quantitative analysis method were established as follows: stationary phase was YMC-Pack ODS-A column (250 mm×4.6 mm, 5 μm, 12 nm) was adopted, and the mobile phase was acetonitrile-water (containing 0.1% phosphoric acid) with gradient elution, the detection wavelength was 220 nm, the column temperature was 30 ℃, the flow rate was 1.0 mL·min^-1. Hierarchical cluster analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied to evaluate the quality of 17 batches of Hanchuang Zupa granules.

Methodological investigation of HPLC fingerprint and content determination were well verified and met the analysis requirements. A total of 25 common peaks were obtained by full peak matching, and eight of them were identified by comparing with the retention time of mixed reference substances. The similarities of 17 batches of samples were above 0.90, which showed good consistency and stability between the samples. Seventeen samples could be classified into three clusters. Three principal components from 21 common peaks were extracted by PCA. Six quality differential compounds were presented in the fingerprints by OPLS-DA, including rutoside, gallic acid, ammonium glycyrrhizinate, chlorogenic acid and so on. The resolution and linear relationship of eight components in quantitative analysis were good. The average recovery rates were 98.0%-99.1% with RSD≤2.0%.

In this study, the qualitative analysis of HPLC fingerprint and quantitative analysis of multiple index components is specific, simple and accurate, which can provide a reference for the quality control and quality evaluation of Hanchuang Zupa granules.

To analyze the fragmentation rule and pathway of pelargonidin, cyanidin, delphinidin, peonidin, petunidin and malvinidol under UHPLC-QTOF-MS positive mode electric spray, identify the characteristic product ions of six anthocyanins, and provide a theoretical basis for the establishment of mass spectrometry database and detection methods.

The chromatographic conditions were as follows: chromatographic column, Fusion-RPC₁₈ (50 mm×2.0 mm, 4 μm), mobile phase 0.1% formic acid aqueous solution (A) and methanol (B), gradient elution (0-1 min, 95%A; 1-5 min, 95%A→10%A; 5-6 min, 1%A; 6-7 min, 95%A), flow rate 0.3 mL·min^-1, column temperature 40 ℃, and injection volume 10 μL. The mass spectrometry conditions were as follows: TOF MS-IDA MS/MS, curtain gas, 0.20 MPa, collision gas CAD 7 MPa, IS voltage, 5 500 V/-4500 V, ion source temperature, 500 ℃, nebulizer gas, GAS1, 0.38 MPa, auxiliary, GAS2, 0.48 MPa, DP voltage, ±60 V, fragmentation voltage, (35±15) V, and time 0.2 s. Under the condition of positive mode of electrospray, the mass spectrometry data of six anthocyanins were measured. According to the pyrylium ions formed by the 2-phenylchromogenic structure of anthocyanins, and combined with the auxiliary analysis of the mass spectrometer database, the possible product ions were deduced.

The results showed that the six anthocyanins mainly undergo cleavage reactions on the pyranium ring, ultimately generating intermediate ions of pyrogallol and benzyl alcohol. On the other way, it occured α cracking, σ cracking causes the loss of functional groups on the ring, ultimately resulting in the formation of Chain hydrocarbons without functional groups.

The research results can provide support for the mass spectrometry characteristic ion data of six anthocyanins, which can be used to establish ion library data for anthocyanin products and also provide reference for the development and research of detection methods.

To develop a flow-through cell method for the dissolution test of omega-3-acid ethyl ester 90 soft capsules and compare the dissolution behaviors from different manufacturers.

The medium (surfactant and its concentration, pH, dosage of pepsin), flow rate and system mode (closed versus open) were investigated. The samples were collected at the specified time and determined by HPLC. The similarity of the dissolution curves between generic drugs and reference listed drug was evaluated by similarity factor (f₂).

A closed-loop mode of flow-through cell apparatus was employed, with 0.01 mol·L^-1 hydrochloric acid solution containing 4.0% Triton X-100 as the dissolution medium, and the flow rate was 2.0 mL·min^-1. The dissolution curves of the samples that have passed consistency evaluation are similar to those of the reference and the samples produced by enterprise in the declaration stage are partly similar. The method has effective distinguish ability for product quality and different prescriptions.

The newly established method can be used for the quality control of omega-3-acid ethyl ester 90 soft capsules, and can provide references for further consistency evaluation and the dissolution method development of lipid-filled soft gelatin capsule (SGC).

To establish an HPLC method for the determination of related substances in nifedipine.

HPLC was adopted on a PFP column (250 mm×4.6 mm, 5 μm) with a gradient elution system of 20 mmol·L^-1 potassium dihydrogen phosphate solution and methanol, the flow rate was 1.0 mL·min^-1, and the column temperature was maintained at 30 ℃. The detection wavelength was set at 265 nm.

The resolutions were good between the peaks of nifedipine and ten known impurities, including impurity D, 2-nitrobenzaldehyde, monoamide, hydroxy dehydro lactone, impurity C, dehydro-N-oxide, impurity A, impurity B, m-nifedipine, p-nifedi-pine. The resolutions between the known impurity peaks were not less than 1.5, the resolutions between the main peak of nifedipine and it’s front and back impurity peaks were not less than 2.0. The calibration curves of mass concentration of above known impurities were linear respectively in their concentration range of 0.000 2-0.015 mg·mL^-1(r>0.999, n=7). The correlation coefficients of above known impurities were 1.000, 1.000, 1.000, 1.000, 0.999 9, 0.999 9, 0.999 9, 1.000, 0.999 9, 0.999 9, respectively. The average recovery rates of above known impurities were 93.1%(RSD=2.3%), 110.6%(RSD=1.9%), 109.2%(RSD=2.0%), 111.0%(RSD=2.1%), 108.1%(RSD=1.9%), 112.4%(RSD=1.8%), 110.8%(RSD=1.9%), 91.5%(RSD=3.1%), 98.9%(RSD=2.7%), 110.1%(RSD=2.6%), respectively. The detection limit of above known impurities was 0.000 06 mg·mL^-1, the quantification limit of above known impurities was 0.000 2 mg·mL^-1. The impurity determination results of the three batches of nifedipine samples showed that the content of the known impurities and the maximum single unknown impurity were less than 0.1%, the total impurities contents were less than 0.5%.

The method has good sensitivity and specificity, and it is suitable for the quality control of nifedipine.

To establish an HPLC method for determination of related substances in apixaban API.

The analytical column was an ACE Excel3 C₁₈-PFP (150 mm×4.6 mm, 3 μm）. The mobile phase A was buffer(30 mmol·L^-1 ammonium acetate in water)-acetonitrile（90∶10） and the mobile phase B was buffer(30 mmol·L^-1 ammonium acetate in water)-acetonitrile（5∶95）. The whole run carried out by gradient elution at a flow rate of 1.2 mL·min^-1. The detection wavelength was set at 280 nm, the column temperature was 40 ℃ and the injection volume was 10 μL.

Apixaban was separated completely from the impurities and degradation products(the resolution>2.0). The test solution was stable for at least 48 h. The LOQs of apixaban, methyl ester product, ethyl ester product, chlorine impurity, dehydrogenation impurity, bihydrolytic impurity, ringopen methyl ester product, cyclate, impurity D, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity, were all 0.05%. The linear correlation coefficients of apixaban, methyl ester product, ethyl ester product, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity were all more than 0.99. The range were from the LOQ for impurity content to 150% of the target concentration. The average recoveries（RSD）(n=9) of methyl ester product, ethyl ester product, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity were 102.0%（2.7%）, 106.4%（2.2%）, 111.2%（4.0%）, 104.4%（2.9%）, 102.9%（2.7%）, 101.8%（2.9%）. The repeatability and intermediate precision completely met the requirements. The impurities contents in three batches of apixaban API 6 months accelerate stability test completely met the requirements, respectively.

This method is simple, rapid, sensitive and specific to be used for the determination of related substances in apixaban API.