Component identification and analysis in vivo of Sanhan Huashi formula / 中国中药杂志

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

Study on fragmentation patterns of coumarins in Notopterygium inchum with ultrahigh performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry / 中国中药杂志

To demonstrate the fragmentation patterns of simple coumarins furanocourmarin(C_7-C_8), furanocourmarin(C_6-C_7) and dihydrofuran coumarin by mass spectrometry, with fraxin, scopoletin, isopsoralen, pimpinellin, isoimperatorin, notopterol and noda-kenin as study subjects, so as to provide a basis for rapid identification of compounds in different subtypes of coumarins. Ultrahigh performance liquid chromatography combined with quardrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was implemented in both positive and negative ion modes. Masslynx software was employed to provide the elemental constituents of each detected ion based on its accurate molecular weight. Chemdraw 2014 was used to cultivate mass number of each inferred structure. The fragment pattern of each compound was determined based on the structures inferred from all the relevant ions. And the patterns were drawn by Chemdraw 2014. The deviation between the calculated molecular weight of the inferred structure and the detected value of the ions was used to assess the correctness of the inferred structures in the fragmentation patterns. The results showed that with UPLC-Q-TOF, neutral loss of CO_2 and CO was reflected in lactone and furan skeletons from the courmarin structure. An even mass was attributed to the loss of an odd number of methyl radicals from compounds with a methoxy substituent. Furanocourmarin(C_7-C_8) produced a protonated molecular ion([M+H]~+), while the other courmarin subtypes produced either a sodium adduct of the molecular ion([M+Na]~+) or a sodium adduct of the molecular ion([M+Na]~+) with a protonated molecular ion([M+H]~+). The m/z 203.03 was a diagnostic ion for furanocourmarin(C_6-C_7), and the m/z 147.04 was supplementary evidence for furanocourmarin(C_6-C_7) identification. The characteristic ion of furanocourmarin(C_7-C_8) was m/z 131.05, while m/z 187.04 was the characteristic ion of dihydrofuran coumarin. The m/z 203.03 ion for furanocourmarin(C_7-C_8) was pretty weak. In negative ion mode, furanocourmarin(C_7-C_8) did not have any signals that were different from the other subtypes of courmarins. The fragmentation patterns in negative ion mode for the other subtypes of courmarins were similar to those in positive ion mode. Four types of fragmentation patterns were identified as forcourmarins from Notopterygium inchum. This study provides the basis for the rapid identification of courmarin subtypes by mass spectrometry.

RP-HPLC fingerprint evaluating different ginger juice as processing material / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method for comparing the differences between fresh and dried ginger juice.</p><p><b>METHOD</b>The RP-HPLC fingerprint method was performed on an Alltech C18 column (4.6 mm x 250 mm, 5 microm) with mobile phase in gradient elution composed of A-acetonitrie and B-water at a flow rate: 0.8 mL x min(-1). The detecting wavelength was 280 nm, and the column temperature 25 degrees C.</p><p><b>RESULT</b>There was no significant difference among the same breed ginger juice of different batches. But there was significant difference between crushed ginger juice and the boiled juice. Trytophan, 6-gingerol were common constituents of the three kinds of ginger juice, the fresh ginger and the dry ginger. Besides, 6-shogaol emerged in the boiled juice.</p><p><b>CONCLUSION</b>The RP-HPLC fingerprints spectrum can be used to distinguish different ginger juices. And the crushed juice of fresh ginger have the same chemical consititents with the fresh ginger.</p>

Study of liquorice processing fructus / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the processing method of fructus evodiae and its standard for quality control, toxicity aspects and pharmacodynamics were carried out at the same time.</p><p><b>METHOD</b>In the studies of processing techniques, the optimized technical parameters were determined by the contents of evodiamine and evodine. And the acute toxicity and pharmacodynamics were studied by rats.</p><p><b>RESULT</b>The process was that the liquorice-processed fructus evodiae was wetted by liquorice decoction by sixth of raw fructus evodiae (V/W) and fried below 230 degrees C. The method of detecting the contents of evodiamine and evodine was that Alltima ODS C18 (4.6 mm x 250 mm, 5 microm); mobile phase acetonitrile-water-tetrahydrofuran-phosphoric acid (51:48: 1: 0.05); column temperature 25 degrees C; mobile rate 0.8 mL x min(-1); wave length 225 nm. The toxicity experimentation show that rats didnt show any notable changes after affused the raw material and the processed fructus evodiae's decotion 40 g x kg(-1) b. w. at one time seven days constantly. The analgesic effect was observed after 0.6 g (material) x kg(-1) (weight) b. w.</p><p><b>CONCLUSION</b>The toxicity of the raw material and the processed one were low and the liquorice-processed fructus evodia analgesic effect was good.</p>

Optimal technique of wine-processed gentian / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize technique of wine-processed gentian (root of Gentiana manshurica, G. scabra, G. triflora and G. rigescens.</p><p><b>METHOD</b>Orthogonal design L9 (3(4)) was used to select the best processing technical parameters by yields of the water extracts and amounts of gentiopicroside in the processing products.</p><p><b>RESULT</b>The optimal procedure was suggested as follows: Gentiana Radix was cut into 5-10 millimetres' long, added one-fifth amounts of wine by gentian's weight, moistened for two hours, and then dried by slow fire.</p><p><b>CONCLUSION</b>The amounts of gentiopicroside in wine-processed gentian were closely related to the types and amounts of wine, moistened time and dried method.</p>

Isolation and structure identification of chemical constituents from the skin of Bufo bufo gargarizans / 药学学报

The skin of Bufo bufo gargarizans, originated from Bufo bufo gargarizans Cantor (Bufonidae), is widely used in traditional Chinese medicine for the treatment of hepatoma, lung cancer and etc. The preparation of the aqueous components has significant therapeutic effect against the digestive tract cancer. The water-soluble chemical constituents in the skin of Bufo bufo gargarizans were then investigated to make clear the active compounds. Six compounds were isolated and purified by recrystallization and column chromatography on silica gel and ODS, their structures were elucidated as 4-amido-3-hydroxymethyl-cyclooctylamidezotetra-alpha-furanone (I), bufogargarizanine C (II), bufothionine (III), dehydrobufotenine hydrobromide (IV), suberic acid (V) and succinic acid (VI) on the basis of physicochemical properties and spectral data (UV, IR, 1H NMR, 13C NMR and MS). Of the above compounds, compounds I and II are new compounds and named bufogargarizanine B and C, respectively.

Methodological studies on equivalent relationship between granule for clinical prescription and clinical decoction of fructus evodiae / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the equivalent relationship between granule for clinical prescription and clincal decoction by use of fructus evodiae as a demonstrated object.</p><p><b>METHOD</b>Compared the equivalent ratio relationship of granule for clincal prescription and clincal decoction by determination of evodiamine, rutaecarpine, evodine, total alkaloids and dried extract ratio as markers, ten batches reference decoctions were prepared according to clinical usage as evaluation standards, common-use processed fructus evodiae products, such as salt-processed fructus evodiae, liquorice-proccesed fructus evodiae, as researching objects, and finally validated by pharmacological trials.</p><p><b>RESULT</b>Equivalent ratios of granule for clical prescription to clincal decoction are about two in all processed products, and the pharmacological evaluation showed no siginificant difference in this ratio.</p><p><b>CONCLUSION</b>This equivalent ratio model could be referenced in the production. But, it must be noticed that different herbal medicines perhaps have different equivalent ratio, which should be studied further according to its techniques and production conditions, and finally need to be revalidated by clinical trial.</p>

RP-HPLC analysis of single, merging, and boiled together Lonicera japonica and Forsythia suspense / 中国中药杂志

<p><b>OBJECTIVE</b>Utilizing RP-HPLC analysis of single, merging and simultaneously boiled Lonicera japonica and Forsythia suspense to find out their identity and difference.</p><p><b>METHOD</b>Samples were separated by Alltech-C18 column (4. 6 mm x 250 mm, 5 microm) with a mixture solution of acetonitrile-water (containing 0.2% ethanoic acid) as mobile phase, 1.0 mL x min(-1) as flow-rate, 280 nm as detected wave-length, room temperature as temperature of column, 10 microL as injected volume.</p><p><b>RESULT</b>The linear ranges of chlorogenic acid and forsythoside A were 0.980-9.800 microg (r = 0.999 9) and 0.972-9. 720 microg (r = 0.999 9), respectively. The average recoveries of chlorogenic acid and forsythoside A were 99.85% (n = 6, RSD 2.0%) and 100.87% (n = 6, RSD 1.6%), respectively. Major peaks were confirmed to the two component herbs, contents of chlorogenic acid and forsythoside A were determinated, ingredients were represented by following peaks were compared qualitatively: peaks k, m, n: L. japonica > merging sample > boiled sample; peaks g, h: Forsythia suspense > merging sample > boiled sample; peaks d, e, i: boiled sample > L. japonica, boiled sample > merging sample.</p><p><b>CONCLUSION</b>Difference and identities among 4 experimental samples of RP-HPLC separations show that Yinqiao, L. japonica and F. suspense' substances are difference and identities, which offered direction for investigating further the topic-pharmacodynamic substance of Yinqiao, provided increasing experimental evidence for clinic medicine and pharmaceutics and supplied reference for the complex prescription of traditional Chinese medicine.</p>

Determination of matrine, sophoridine and oxymatrine in Compound Kushen Injection by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a high performance liquid chromatography (HPLC) method for determination of matrine, sophoridine and oxymatrine in Compound Kushen Injection.</p><p><b>METHOD</b>Alltima amino column was used with acetonitrile-3% phosphoric acid-ethyl alcohol (80:10:10) as the mobile phase and the detection wavelength was at 220 nm.</p><p><b>RESULT</b>The mean recovery of matrine, sophoridine and oxymatrine in Compound Kushen Injection was 99.51% (RSD 1. 58%), 99.24% (RSD 1.44%) and 100.22% (RSD 1.85%), respectively.</p><p><b>CONCLUSION</b>The method was simple, rapid, accurate and specific and suitable for quality control of the product.</p>

Determination of aristolochic acid A in Guanxinsuhe preparations by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a determination method of aristolochic acid A in Guanxisuhe preparations by RP-HPLC.</p><p><b>METHOD</b>The instrument used was Hewlett-Packard 1100 HPLC with a Alltech C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase was methanol-water-acetic acid (68: 32:1) and the flow rate was 1.0 mL x min(-1). The UV detection wavelength was 390 nm and the column temperature was at 35 degrees C. The extracted solvent for the preparations was methanol solution contained 10% formic acid.</p><p><b>RESULT</b>The calibration curve was linear (r = 0.999 9) within the range of 0.119-1.89 microg for aristolochic acid A. The average recovery 99.0%, RSD 0.63%.</p><p><b>CONCLUSION</b>The method with good linear relationship was convenient, quick, accurate, and suitable for the quality control of the aristolochic acid A in Guanxinsuhe and other traditional Chinese medicines containing aristolochic acid A.</p>

Multi-components quantitation by one marker new method for quality evaluation of Chinese herbal medicine / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a new quality evaluation method for Chinese herbal medicine, using one chemical reference substance to calculate multi-components simultaneously.</p><p><b>METHOD</b>Relative correction factors (RCF) of mutongsaponin B and C to saponin PJ1, a characteristic component as marker, were calcalated in The chromatographic conditions for determination of the three saponins in Caulis Akebiae akebiae as a research model. The contents of saponin PJ1 were determined by external standard method, and those of mutongsaponin B and C were calculated by saponin PJ1 and their RCF. The accuracy of the new method was evaluated by comparing the calculated contents with the determined contents.</p><p><b>RESULT</b>The analysis procedures were validated. There has been no significant difference between the calculated contents and the determined contents, according to the angle cosine value, and the calcalated RCF have confidence value.</p><p><b>CONCLUSION</b>The method of multi-components quantitation by one marker has been verified in caulis akebiae and it is to be a new quality evaluation pattern for Chinese herbal medicines.</p>

Determination of acetylharpagide in Ajuga decumbens by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method of the quantitative determination of acetylharpagide in Ajuga decumbens.</p><p><b>METHOD</b>The chromatographic conditions were as follows: a Phenomenex Luna C18 column was used, the mobile phase was composed of acetontrile-water (15:85), the flow rate was 1.0 mL x min(-1) and the UV absorbance detection was set at 197 nm.</p><p><b>RESULT</b>Linearity of acetylharpagide was in the range of 0.6-3.6 microg (r = 0.9993), and the average recovery and RSD were 99.13% and 2.48%, respectively.</p><p><b>CONCLUSION</b>The contents of acetylharpagide ranged 0.40%-6.39% in A. decumbens. The method was simple, accurate and sensitive.</p>

Methodological studies on selectively removing toxins in Aristolochiae manshuriensis by chinese processing techniques / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the possibility of selectively detoxifying aristolochic acids in Aristolochiae manshuriensis (Guanmutong) by the chemical properties of aristolochic acids and traditional Chinese processing experience.</p><p><b>METHOD</b>The technical parameters in processing technique of A. manshuriensis were optimized by orthogonal designed methods with aristolochic acid A.</p><p><b>RESULT</b>The processing technique was soaked in 0.1 mol x L(-1) baking soda for several times and then processing with vinegar. Temperature was important factor to detoxify aristolochic acids. aristolochic acid A were removed over 90% from A. manshuriensis in laboratory and over 80% in medium-scale production by 10 batches from two origins of botanical drugs with different contents, and decreased to 0.35-0.60 mg x g(-1) in processed products. aristolochic acid A existed mostly salt-forms in the botanical drug.</p><p><b>CONCLUSION</b>Most toxic components in Guanmutong could be removed by the new processing method.</p>

Studies of commonly used traditional medicine-ginger / 中国中药杂志

To review the chemical, pharmacological, studies on ginger (Zingiber officinale) in the last ten years, and also its processing history and clinical uses. Gingerols and related compounds in ginger have many pharmacological activities. Chemical studies should be given sufficiently emphasis, and advances of the chemical study will promote the other related researches to develop in depth.

HPLC determination of 6-gingerol in Rhizoma Zingiberis Recens / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the contents of 6-Gingerol in Rhizoma Zingiberis Recens.</p><p><b>METHOD</b>HPLC method was used, with Alltech C18 column, acetonitrile-methol-water (43:5:52) as mobile phase with a flow rate of 0.8 mL.min-1, detecting wavelength 280 nm, and column temperature 35 degrees C.</p><p><b>RESULT</b>Retained time of 6-gingerol was near 19 min, showing a good recovery (98.2%) and linear correlation (r = 0.9999). The contents of 6-gingerol were 1.35-2.87 mg.g-1, and the water contents were 70.4-85.5% mL.g-1 in Rhizoma Zingiberis Recens.</p><p><b>CONCLUSION</b>The method is appropriate for the determination of 6-gingerol in Rhizoma Zingiberis Recens. Gingerol can be used as a chemical marker of the quality control of Rhizoma Zingiberis Recens.</p>