Berberine Content Research Articles

Optimization of extraction of bioactive alkaloids and antioxidants from rhizome coptidis (Coptis Chinensis Franch) by using response surface methodology

The optimum conditions for the extraction of bioactive alkaloids and antioxidants from rhizome coptidis were determined by using response surface methodology (RSM). A small central composite design (SCD) was employed to investigate the effect of 3 process variables such as extraction time (3–7 h), ethanol concentration (0–100%), and solvent to sample ratio (25–45 mL/g) during extraction process. Analysis of variance showed a high significance of fitted model which indicated the model is reliable for prediction. The optimal conditions for alkaloids and antioxidant ability were obtained using ridge analysis, and under the optimum conditions the corresponding predicted response values for berberine content, palmatine content, and antioxidant ability were 747.2 μg berberine chloride equivalent/mL, 168.34 μg palmatine chloride equivalent/mL, and 2.99 mmol/L Trolox equivalent, respectively, which well agreed with predicted values, indicating the success of RSM in optimizing the extraction conditions.

과산화수소/자외선/산소 처리를 이용한 베르베린 염료 및 염직물의 퇴화거동 연구

Abstract This study examined the degradation behavior of SB(standard berberine) dye and SB dyed silk using HPLC-MS instrument after degradation in the hydrogen peroxide/ultraviolet ray radiation/oxygen system up to 9 days and 40 hours respectively. In the degraded samples, berberine was detected at 5.2 min in the SB dye and 5.3 min in the SB dyed silk with its molecular ion=336 and the UV spectra of quaternary alkaloid. Degradation product 3(m/z=102) newly appeared after 5 day degradation treatment with continued increase till the end of degradation treatment. The amount of berberine in the degraded dye decreased with degradation progression. In the silk dyeings, berberine was detected only up to 21 hour degradation sample. The amount of berberine decreased dramatically during the first 6 hours of degradation treatment. The CIELAB color measurement of the silk dyeings showed dramatic change in the b* value, near zero in the 40 hour degraded silk. CIELAB and Munsell color measurements were in agreement with the HPLC-MS results of the dyed silk in the change of berberine content that the degraded silk became white and lost yellow color.Keywords: berberine(베르베린), degradation(퇴화), Phellodendron bark(황백), HPLC-MS(고속 액체 크로마토그라피 질량분석), color measurement(측색)

Standardization of Berberis aristata extract through conventional and modern HPTLC techniques

ObjectiveBerberis aristata (Berberidaceae) is an important medicinal plant, found in the different region of the world. It has significant medicinal value in the traditional Indian and Chinese system of medicine. The aim of the present investigation includes qualitative and quantitative analysis of Berberis aristata extract. MethodsPresent study includes determination of phytochemical analysis, solubility test, heavy metal analysis, antimicrobial study and quantitative analysis by HPTLC method. ResultsPreliminary phytochemical analysis showed the presence of carbohydrate, glycoside, alkaloid, protein, amino acid, saponin, tannin and flavonoid. Solubility in water and alcohal were found to be 81.90% in water and 84.52% in 50% in alcohal. Loss on drying was found to be 5.32%. Total phenol and flavonoid content were found to be 0.11% and 2.8%. Level of lead, arsenic, mercury and cadmium complies the standard level. E. coli and salmonella was found to be absent whereas total bacterial count, yeast and moulds contents were found to be under the limit. Content of berberine was found to be 13.47% through HPTLC techniques. ConclusionsThe results obtained from the present studies could be used as source of valuable information which can play an important role for the food scientists, researchers and even the consumers for its standards.

Anti-diabetic effect of a traditional Chinese medicine formula

An anti-diabetic TCM formula consisting of Schizandra chinensis Baill. (SC), Coptis chinensis (CC), Psidium guajava L. leaves (PG) and Morus alba L. leaves (MA) was developed based on its α-glucosidase, DPP-4 and AGE inhibitory activities in vitro using response surface methodology (RSM). Then, the in vivo study was carried out to confirm the anti-diabetic function of the formula. RSM results showed that the optimum anti-diabetic TCM formula is the combination SC (3000 μg mL(-1)), CC (80 μg mL(-1)), PG (374.56 μg mL(-1)) and MA (480 μg mL(-1)). For the in vivo study, insulin resistant mice were induced by high-fat/high-sucrose (HF/HS) feeding for 6 weeks. Administration of the developed formula significantly decreased non-fasting blood glucose in the HF/HS diet mice. Moreover, the formula decreased blood glucose levels in the insulin tolerance test. These results indicated that the anti-diabetic mechanism of the formula might be due to decreased insulin resistance. The serum fructosamine level in the high dose group was significantly lower than the HF/HS and normal control groups, indicating that the formula could improve middle term glucose levels and reduce the risk of complications. The contents of berberine and 1-deoxynojirimycin in the formula were 4.7 ± 0.4 and 77.1 ± 1.1 μg mL(-1), respectively. These two compounds can be used as indicators for quality control during production.

Determination of coptis chinensis’ quality by FT-NIR spectroscopy

As a kind of traditional Chinese medicinal materials coptis chinensis has a lot of medicinal value and closely related to human health, Widely used in clinical or disease prevention and treatment. So the quality of medicinal coptis chinensis rapid determination is very important. Calibration model for determination of Coptis chinensis’ quality was built by partial least squares and near infrared spectroscopy, and the method of spectral pretreatment was discussed. Berberine and moisture content were used as evaluation index of Coptis chinensis’ quality according to traditional Chinese medicine standard in this experiment. The first derivative and vector normalization were adopted for the spectral preprocessing. The performance of the final model was evaluated according to root mean squares error of cross-validation (RMSECV), root mean square error of prediction (RMSEP) and correlation coefficient (R). According to the model of calibration set, the R values were 0.9298 and 0.8898, RMSECV were 0.239% and 0.302%. The R values of prediction set were 0.9347 and 0.9146, RMSEP were 0.104% and 0.330%, and prediction results were accurate and reliable. This method could be applied in rapid analysis of Coptis chinensis. The method was suitable for production fields and research fields, and had a wide application prospect.

Improvement of Benzylisoquinoline Alkaloid Productivity by Overexpression of 3′-Hydroxy-<i>N</i>-methylcoclaurine 4′-<i>O</i>-Methyltransferase in Transgenic <i>Coptis japonica</i> Plants

Coptis japonica (Cj) rhizomes are used as a crude drug for gastroenteritis, since they accumulate antimicrobial berberine. Berberine also shows various useful bioactivities, including cholesterol-lowering activity. Unfortunately, Cj is a slow-growing plant and more than 5 years are required to obtain a crude drug suitable for the Japanese Pharmacopoeia. To improve alkaloid productivity, we overexpressed the 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4'OMT) gene in Cj. We established the transgenic plant (named CjHE4') by introducing one copy of Cj4'OMT by Agrobacterium-mediated transformation. The successful overexpression of 4'OMT was confirmed in all tissues of CjHE4' by real-time polymerase chain reaction (PCR) analysis. HPLC analysis revealed that the berberine content of CjHE4' leaves and roots cultivated for 4 months was increased to 2.7- and 2.0-fold, respectively, compared with non-transgenic wild-type (CjWT), and these inductions of alkaloids were stable for at least 20 months. Furthermore, in CjHE4' cultivated for 20 months, the berberine content in medicinal parts, stems and rhizomes was significantly increased (1.6-fold). As a consequence, increased amounts of alkaloids in CjHE4' resulted in the improvement of berberine yields (1.5-fold), whereas CjHE4' showed slower growth than CjWT. These results indicated that 4'OMT is one of the key-step enzymes in berberine biosynthesis and is useful for metabolic engineering in Cj.

Effect of enhanced UV-B radiation on metabolism and berberine content of Coptis chinensis

To reveal the response of content berberine in root of Coptis chinensis to different intensity of UV-B radiation, and provide the theory basis for promoting the content of berberine. Four groups of UV-B radiation were set in the experiment which included: natural light control (0 W x m(-2)), UL (0.05 W x m(-2)), UM (0.10 W x m(-2)), UH (0.20 W x m(-2)). The special photosynthesis character, PPP pathway in the primary metabolism and lyrosinase activity, the changes of berberine in the root of C. chinensis were measured under different UV-B radiation. Photosynthetic pigment, qN, Fo, ETR, activity of glucose-6-phosphate dehydrogenase and the content of berberine in the root of C. chinensis, all of these parameters were lower than other groups under the UH radiation. However, under the UM radiation, C. chinensis protected itself from the light UV-B radiation by promoting the power of photosynthesis and PPP pathway in order to produce more NADPH and secondary metabolites. C. chinensis increases its photosynthetic ability and PPP pathway which can furnish more precursor of secondary metabolites and NADPH that are needed in the secondary metabolism. Furthermore, the content of berberine increases correspondingly. The research provide the example for increasing the content of berberine in C. chinensis cultivation.

황벽 추출염료와 염직물의 열적 퇴화 거동 연구

This research compares the change in berberine content in liquid dye and the color and dye uptake of amur cork tree dyed silk upon thermal degradation treatment. Thermal degradation of amur cork tree extract and liquid dye of standard berberine was carried out at room temperature, <TEX>$4^{\circ}C$</TEX> refrigeration, and <TEX>$100^{\circ}C$</TEX> oven conditions for 0-192 hours. Amur cork tree dyed silk was treated in a <TEX>$100^{\circ}C$</TEX> oven for 0-240 hours. The berberine content in liquid dye was measured by the relative abundance of the berberine peak in the HPLC-MS chromatograms. The color and dye uptake of dyed silk was measured using K/S value and colorimetric data. The berberine content in standard berberine dye was 2.4 times that of the amur cork tree extract. A similar result was observed between the K/S value of standard berberine dyed silk and that of amur cork tree dyed silk. The berberine dyed silk showed the highest dye uptake after 120 hours in a <TEX>$100^{\circ}C$</TEX> oven. This result was similar to the change in a berberine content in liquid dye in a <TEX>$100^{\circ}C$</TEX> oven treatment. The change of the K/S value of amur cork tree dyed silk and berberine content of amur cork tree extract was similar up to 24 hours. The result suggests that there is a direct relationship between the color change of amur cork tree dyed silk and the berberine content in amur cork tree dye.

Correlation analysis soil nutrition as well as yield and active compounds of Coptis teeta

The correlations between soil nutrition and yield as well as active compound contents of Coptis teeta were analyzed. The contents of 13 soil factors, rhizome biomass and the content of berberine, jatrorrhizine, and palmatine in rhizome of C. teeta were determined, and analyzed by factor analysis (FA) and correlation analysis (CA). FA showed that the first three factors accounted for 86.9% of the total variance. The contents of Ca, Mg, Fe, Mn, and Zn correlated with the first factor. The content of organic matter, N, available N, available P, and available K correlated with the second factor. CA showed that yield of C. teeta had a significant positive correlation with available P (r = 0.931) at 0.01 level, and available N (r = 0.703) at 0.05 level. The content of berberine of C. teeta had a significant positive correlation with available P (r = 0.680) at 0.05 level. The yield and berberine content of C. teeta were high in the soil with high contents of available N and available P.

Influence of Environmental Factors on Production of Berberine Content in Berberis asiatica Roxb. ex DC in Kumaun West Himalaya, India

Berberis asiatica samples collected from nine altitudes of Kumaun Himalaya, India were analyzed for berberine concentrations using high-performance liquid chromatography. Correlation between berberine concentration, size class, and season were established. The root and stem bark samples from lower altitude, having larger size class, contained more berberine. Berberine concentration was the highest in summer and lowest in the rainy season. Low level of soil moisture and higher level of soil potassium produced higher berberine concentrations.

Comprehensive Study in the Inhibitory Effect of Berberine on Gene Transcription, Including TATA Box

Berberine (BBR) is an established natural DNA intercalator with numerous pharmacological functions. However, currently there are neither detailed reports concerning the distribution of this alkaloid in living cells nor reports concerning the relationship between BBR's association with DNA and the function of DNA. Here we report that the distribution of BBR within the nucleus can be observed 30 minutes after drug administration, and that the content of berberine in the nucleus peaks at around 4 µmol, which is twelve hours after drug administration. The spatial conformation of DNA and chromatin was altered immediately after their association with BBR. Moreover, this association can effectively suppress the transcription of DNA in living cell systems and cell-free systems. Electrophoretic mobility shift assays (EMSA) demonstrated further that BBR can inhibit the association between the TATA binding protein (TBP) and the TATA box in the promoter, and this finding was also attained in living cells by chromatin immunoprecipitation (ChIP). Based on results from this study, we hypothesize that berberine can suppress the transcription of DNA in living cell systems, especially suppressing the association between TBP and the TATA box by binding with DNA and, thus, inhibiting TATA box-dependent gene expression in a non-specific way. This novel study has significantly expanded the sphere of knowledge concerning berberine's pharmacological effects, beginning at its paramount initial interaction with the TATA box.

Study on the Electrochemical Behavior of the Anticancer Herbal Drug Berberine and Its Analytical Application

AbstractThe electrochemical behavior of anticancer herbal drug berberine was investigated in 0.2 mol L‐1 HAc‐NaAc (pH 4.2) buffer solution at a glassy carbon electrode. The drug yields a single well‐defined reduction peak at the potential of ‐1.085 V at the scan rate of 0.05 V s‐1 and the electrode reaction is an irreversible adsorption‐controlled process as the result of cyclic voltammetry. All experimental conditions were studied in order to obtain the optimum conditions for the determination of berberine. A good linear relationship was observed between the reductive peak currents and the concentrations of berberine in the range from 1.0 × 10‐6 to 8.0 × 10‐5 mol L‐1. The detection limit of 8.0 × 10‐7 mol L‐1 was obtained in terms of the signal to noise ratio of 3:1 (S/N = 3). The proposed technique was applied to determine the content of berberine in tablets with good satisfactory. In addition, the electrode process dynamics parameters were calculated.

Quantitative analysis of berberine inBerberis aristatafruits and in a traditional anti-inflammatory unani formulation by use of a validated HPLC method

Summary A validated reversed-phase HPLC method has been developed for quantitative analysis of berberine in Berberis aristata fruits and in a polyherbal formulation. Separation of berberine was achieved on a C18 column with a mobile phase consisting of a 10–80% acetonitrile gradient in 0.05% aqueous orthophosphoric acid. The flow rate was 1 mL min−1. Detection was at 266 nm. A sharp, well defined peak was obtained at a retention time of 10.0 ± 0.4 min. The method was validated in accordance with ICH guidelines for accuracy, precision, robustness, and the limits of detection (LOD) and quantification (LOQ). Results from linear regression analysis were indicative of a good linear relationship (r2 = 0.998 ± 0.0011) in a wide concentration range (5–500 μ g mL−1). LOD and LOQ were 1.5 and 5.3 μg mL−1, respectively. Satisfactory recovery results (94.6–103.1%) were obtained by the method of standard addition. Intra-day, inter-day, and intersystem precision was satisfactory, with relative standard deviation in the range 0.7–1.8%. The berberine content of fruit of Berberis aristata and the herbal formulation were 0.033% and 0.0089% (w/w), respectively. This HPLC method for quantification of berberine can be used for quality control and standardization of several crude drugs and different herbal formulations in which berberine is present as a phyto constituent.

The alteration of components in the fermented Hwangryunhaedok-tang and its neuroprotective activity.

Background:Hwangryunhaedok-tang is a traditional herbal prescription that has sedative activity, hypotensive and anti-bacterial effects.Objective:In this study, we investigated the alteration of contents of components in Hwangryunhaedok-tang, antioxidant activity and neuroprotective activity by fermentation with Lactobacillus acidophilus KFRI 128.Materials and Methods:Contents of three marker compounds (geniposide, berberine and palmatine) and unknown compounds in the Hwangryunhaedok-tang (HR) and the fermented Hwangryunhaedok-tang (FHR) were measured and compared using the established high-performance liqued chromatograph coupled with a photodiode (HPLC-DAD) method. The antioxidant activity of HR and FHR were determined by DPPH free radical and hydrogen peroxide (H2O2) scavenging assay. Also, the neuroprotective activities of HR and FHR against glutamate-induced oxidative stress in a mouse hippocampal cell line (HT22) were evaluated by MTT assay.Results:The contents of geniposide and palmatine were decreased but the content of berberine was increased in the FHR. And the contents of unknown compounds (1), (2), (3), (4) and (5) in the HR were altered by fermentation. Electron donating activity (EDA, %) value of FHR was higher than HR for DPPH radical scavenging activity and H2O2 scavenging activity, respectively. In the MTT assay, FHR showed more potent neuroprotective activity than HR by 513.90%.Conclusion:The FHR using microorganism could convert compounds in HR and enhance the antioxidant and neuroprotective activity.

Analysis of Berberine Content using HPTLC Fingerprinting of Root and Bark of Three Himalayan Berberis Species

Analysis of Berberine Content using HPTLC Fingerprinting of Root and Bark of Three Himalayan Berberis Species

Preliminary Studies Toward Identification of Sources of Protoberberine Alkaloids used as Yellow Dyes in Asian Objects of Historical Interest

The protoberberines bright yellow alkaloids that have been used for centuries in Chinese and other traditional medicines and also as dyes for textiles and paper. The most frequently cited source for protoberberine dyes is the Amur cork tree (phellodendron amurense). However, many plants contain protoberberines, and, as shown here, the analysis of dyed objects often permits identification of plant types, which sheds light on the resources available to early dyers. In this study, representatives of five genera (Berberis, coptis, Fibraurea, Mahonia, phellodendron) of protoberberine-yielding plants were analyzed, all of which have been reported as having been used in Asia as sources of yellow colorants, using high-performance liquid chromatography (HPLC) with diode array and mass spectrometric detection. It is shown that their contents of berberine, palmatine, jatrorrhizine and other protoberberines are distinctive enough to allow them to be distinguished from one another. As a result, analysis of the dyes used in several historical objects allows conclusions to be drawn as to the plant source of the dye. Knowledge of the type of colorant used in an object is important for its conservation because the protoberberine alkaloids, as a class, are both very water-soluble and relatively unstable (among yellow dyes) to light.

High-Performance Thin Layer Chromatography for Quality Control of Multicomponent Herbal Drugs: Example of Cangzhu Xianglian San

Due to their complexity, multicomponent herbal drugs pose enormous analytical challenges for quality control (QC). Although they may have traditionally been used for hundreds of years, the information about their chemical composition is often still limited. Selecting suitable markers to monitor the identity and potency of the mixture is, therefore, difficult. There is also the possibility of natural variability for each plant. This paper illustrates a pragmatic and practical approach to QC of a multicomponent herbal drug by HPTLC. Cangzhu xianglian xan (CXS), composed of the herbal drugs Coptis rhizome, Aucklandia root, and Atractylodes rhizome (30 + 20 + 60, w/w/w), is used as an example. A characteristic fingerprint can be generated for CXS with toluene-ethyl acetate-methanol-isopropanol-water (60 + 30 + 20 + 15 + 3, v/v/v/v/v) mobile phase on HPTLC silica gel 60 conditioned with ammonia. While the corresponding monograph of the Chinese Veterinary Pharmacopoeia focuses only on the detection of berberine, one of the principal components of Coptis rhizome, the proposed method of identification determines the presence of all three components in the drug after derivatization with anisaldehyde reagent. The same method can also be used to quantitatively determine the content of berberine by scanning densitometry. This paper provides details about the validation of the qualitative and quantitative determinations.

Simultaneous determination of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex by RP-HPLC

To develop a HPLC method to determine the content of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex simultaneously. MEHTOD: The separations were carried out at 25 degrees C on a Phenomenex Gemini C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril and water containing 0.1% phosphoric acid in gradient mode. The flow rate was 1.0 mL x min(-1), detection wavelengthes were 345 nm for jatrorrhizine , palmatine, berberine and 210 nm for obacunone. The average recoveries of jatrorrhizine, palmatine, berberine, and obacunone were 98.94%, 101.17%, 96.22% and 98.90%, respectively. The method is simple, accurate and repeatable, and can be used in content determination of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex.

Anti-herpes simplex virus effects of berberine from Coptidis rhizoma, a major component of a Chinese herbal medicine, Ching-Wei-San

Berberine is an alkaloid extracted from Coptidis rhizome. Among the individual herbal components of a Chinese herb medicine, Ching-Wei-San, Coptidis Rhizoma has the most potent antimicrobial activity. By high-pressure liquid chromatography, the quantitative analysis of berberine from 6.25-mg/mL (w/v) Coptidis rhizome extract or 50.00-mg/mL (w/v) Ching-Wei-San was determined to be 0.26mg/mL. To explore the potential use of Ching-Wei-San against herpes simplex virus (HSV) infection, the cytotoxicity, anti-HSV-1 and anti-HSV-2 activity in Vero cells were assayed. The selectivity index of berberine was about 1.2-1.5 times higher than that of Coptidis rhizome extract and Ching-Wei-San. Moreover, the antiviral activities correspond to the content of berberine in the aqueous solution. Berberine may interfere with the viral replication cycle after virus penetration and no later than the viral DNA synthesis step, and its activities were not affected by the preparation processes. Berberine, the natural plants that contain this component, including Coptidis rhizome, and Ching-Wei-San have all shown anti-HSV effects.

Metabolism,transformation and distribution of Coptis chinensis total alkaloids in rat

To determine the pharmacokinetics, distribution and mutual transformation of the total alkaloids, jatrorrhizine, coptisine, berberine and palmatine from Coptis chinensis in rats. After the total alkaloids and berberine were fed into rats, their contents in plasma, tissues and gastrointestinal tract were determined by reversed-phase HPLC. The peak times of berberine in blood were 2.0 h (Cmax 3.7 mg x L(-1)) and 5.0 h Cmax 2.8 mg x L(-1)), respectively. Berberine in rat blood can be transformed into jatrorrhizine. After the rats were fed with the total alkaloids by gavage, the content of berberine was decreased monotonously, while coptisine, palmatine and jatrorrhizine contents were increased gradually in the stomach, it speculated that berberine may be transformed into jatrorrhizine in the stomach. Animal experiments showed that berberine and palmatine were mainly distributed in the lungs of animals, followed by the distribution in the liver, while jatrorrhizine and coptisine was mainly in the liver, then in the lungs. Berberine could transform into jatrorrhizine. The mechanism on the appearance of two maximum blood concentration of berberine in blood could be explained with the propulsion of the gastrointestinal tract partly.