Development and evaluation of a herbal formulation with antipathogenic activities and probiotics stimulatory effects

Searching alternative feed antibiotics is always a study hotspot in the field of animal production. In this study, the antipathogenic activities and probiotics stimulatory effects of 30 kinds of herbs were screened through Oxford cup method and Hungate roll method. 15 herbs showed significantly antibacterial activities（P〈0.05） against Escherichia coli, Salmonella enteritidis, Salmonella typhimurium and Staphylococcus aureus. Seven herbs showed greatly stimulatory promoting effects on Lactobacillus acidophilus and Bifidobacterium longum. Finally, five herbs were combined for the developed formulation with selective antibacterial properties and they were Mume Fructus（35%）, Isatidis Folium（25%）, Moslae Herba（20%）, Chrysanthemi Indici Flos（13%） and Bupleuri Radix（7%）. The herbal formulation showed significantly antibacterial abilities against four pathogens and stimulatory promoting abilities on two probiotics in vitro and the equivalent activities in broiler chickens in vivo against E. coli and L. acidophilus. The toxicity study showed it had no toxicity, which indicated that it would be a kind of preferred candidate for an alternative antibiotic in future animal production.

Estimation of boswellic acids in herbal formulations containing Boswellia serrata extract and comprehensive characterization of secondary metabolites using UPLC-Q-Tof-MS^e

Boswellia serrata is a widely used herb in Indian systems of medicine and is well known for its potential medicinal properties.A chromatographic method was developed for the analysis and quanti cation of six boswellic acid marker compounds,i.e.,keto boswellic acid(1),3-O-Acetyl 11-keto-boswellic acid(2),-b Boswellic acid(3),-Boswellic acid(4),3-O-Acetyl--boswellic acid(5)and 3-O-Acetyl--boswellic acid b(6)in commercial herbal products containing B.serrata as an ingredient.Combining UPLCbwith Q-Tof-MS/MS makes the better identi cation of secondary metabolites and adulterants in the herbal formulations containing B.serrata in rapid time using fragmentation approach than the traditional approaches.In this study quanti cation of boswellic acids with UPLC-PDA method was performed as per the pharmacopeia guidelines.Furthermore,minor phytochemical constituenBoswellia serrata is a widely used herb in Indian systems of medicine and is well known for its potential medicinal properties.A chromatographic method was developed for the analysis and quantification of six boswellic acid marker compounds,i.e.,keto boswellic acid(1),3-O-Acetyl 11-keto b-boswellic acid(2),ɑ-Boswellic acid(3),b-Boswellic acid(4),3-O-Acetyl-ɑ-boswellic acid(5)and 3-O-Acetyl-b-boswellic acid(6)in commercial herbal products containing B.serrata as an ingredient.Combining UPLC with Q-Tof-MS/MS makes the better identification of secondary metabolites and adulterants in the herbal formulations containing B.serrata in rapid time using fragmentation approach than the traditional approaches.In this study quantification of boswellic acids with UPLC-PDA method was performed as per the pharmacopeia guidelines.Furthermore,minor phytochemical constituents were identified and characterized with the help of LC-Q-Tof-MS/MS fragmentation data and various isoforms of boswellic acids and tirucallic acids in B.serrata oleo-gum-resin extract were identified.ts were identi ed and characterized with the help of LC-Q-Tof-MS/MS fragmentation data and various isoforms of boswellic acids and tirucallic acids in B.serrata oleo-gum-resin extract were identi ed.

Evaluation of Toxicological Profile of a Polyherbal Formulation

In current study toxicological profile of a commonly used herbal formulation was evaluated that is used extensively for gynecological disorders like menorrhagia, metrorrhagia, leucorrhea, irregular menstrual cycle, pre-menstrual syndrome and post-menopausal bleeding. It was also claimed to strengthen endometrium and ovaries. Since this herbal formulation was been used by a large number of population hence there was a need to assess acute and sub-chronic toxicity. Acute oral toxicity (LD50) was observed in albino mice using standard protocols whereas sub-chronic, hematological and histopathological studies were assessed on 24 albino rabbits after giving herbal formulation for 60 days in two doses (20 and 60 mg/kg) against control groups. The outcomes of present study showed that the drug is safe up to 5000 mg/kg following acute oral toxicity test and no mortality was observed during sub chronic toxicity studies. Results of sub-chronic toxicity did not show any significant changes in biochemical, hematological and histopathological parameters. However, some indicators such as urea, creatinine, hemoglobin, and RBC count were altered, but these changes do not correlate with the histopathological results and may be associated to intra individual variations. Despite the safety of the drug in few animals, clinical trials and more investigations on a large number of animals are essentially needed to establish safety and efficacy of the herbal formulation.

Inhibition of Metalloproteinase Activity by Chinese Formulations Used to Treat Inflammatory Diseases

Matrix metalloproteinases （MMPs） are known to be involved in a number of pathological processes including cancer, atherosclerosis, arthritis and neurodegenerative disease among others. The drive to develop MMP inhibitors as therapeutics has been put forward for years by pharmaceutical companies as well as academicians. In an attempt to screen for MMP inhibitors from Traditional Chinese Medicines （TCMs） , a number of Chinese formulations used to treat inflammatory diseases such as nephritis and hepatitis have been studied. Strong inhibitory effects of three Chinese formulations toward the activity of MMP-16 have been discovered. These results suggest that these anti-inflammatory medicines contain some unknown MMP inhibitory compound（s） and provide reasonable molecular mechanisms for their theraoeutic effects.

Quantitative estimation of (-)-hinokinin, a trypanosomicidal marker in Piper cubeba, and some of its commercial formulations using HPLC-PDA

The fruits of Piper cubeba have been used in Ayurvedic system of medicine for pain,tastelessness, painful urination and mouth diseases. Among its various chemical constituents,（-）-hinokinin, a trypanosomicidal dibenzylbutyrolactone lignan, is found in significant quantities.For quality evaluation of P. cubeba fruit and its commercial formulations, there is an urgent need to develop an analytical method based on（-）-hinokinin. For this purpose, an HPLC method was developed using photo diode array detector and Waters HR C18 column with gradient elution consisting of water and acetonitrile. The developed method was validated as per ICH-Q2 B guidelines and found to be accurate, precise and linear over a wide range of concentrations（5–300 mg/m L）.（-）-Hinokinin contents were found to be in the range of 0.005–0.109%（m/m） in various P. cubeba samples. The developed method was extended to LC–MS for further identification and characterization of（-）-hinokinin in samples. The developed method is simple,rapid and specific, and can be used as a tool for quality control of P. cubeba fruits and its commercial formulations.

Chinese herbal medicine and COVID-19: quality evaluation of clinical guidelines and expert consensus and analysis of key recommendations

Objective:To systematically review the clinical practice guidelines(CPGs)for the treatment of patients with coronavirus disease 2019(COVID-19)using Chinese herbal medicine(CHM),assess the methodological quality as well as clinical credibility and implementability of specific recommendations,and summarize key recommendations.Methods:As of April 2022,we conducted a comprehensive search on major electronic databases,guideline websites,academic society websites,and government websites to assess the methodological quality and clinical applicability of the included CPGs using the Appraisal of Guidelines for Research and Evaluation(AGREE)II tool and Evaluation-Recommendations EXcellence(AGREE-REX)instructions,respectively.Results:The search yielded 61 CPGs,which were mostly published in 2020;moreover,98.4%of the CPGs were published in China.Only five CPGs achieved a high-quality AGREE II rating;further,six CPGs could be directly recommended,with most of the CPGs still showing much room for improvement.CPGs had a low overall score in the AGREE-REX evaluation,with the domains of clinical applicability,values and preferences,and implementability being standardized in 21.80%±12.56%,16.00%±11.81%,and 31.33%±14.55%of the CPGs,respectively.Five high-quality CPGs mentioned 56 Chinese herbal formulas.Half of the recommendations had moderate or strong evidence level in the GRADE evaluation.The most frequently recommended herbal medicines were Lianhua Qingwen granule/capsule and Jinhua Qinggan granule;however,the strength of recommendation for each prescription varied across CPGs and populations.Conclusions:The overall quality of current CPGs for COVID-19 for CHM still needs to be improved;moreover,the strength of the evidence remains to be standardized across CPGs.

A Systemic Review on Topical Marketed Formulations, Natural Products, and Oral Supplements to Prevent Androgenic Alopecia: A Review

Androgens have an intense consequence on the human scalp and body hair.Scalp hair sprouts fundamentally in awol of androgens whereas the body hair hike is vulnerable to the activity of androgens.Androgenetic alopecia(AGA)invoked as males emulate Alopecia due to the cause of the dynamic reduction of scalp hair.Androgens are medium of terminus growth of hair although the body.Local and system androgens convert the extensive terminal follicles into lesser vellus like structure.The out start of this type of alopecia is intensely irregular and the reason behind this existence of enough circulating steroidal hormones androgens and due to genetic predisposition.Effective treatments are available in the market as well as under clinical and preclinical testing.Many herbal formulations are also available but not FDA approved.Different conventional and NDDS formulations are already available in the market.To avoid various systemic side effects of both Finasteride and Minoxidil,topical formulations and natural products(nutrients,minerals,vitamins)now a days are being widely used to treat Androgenic alopecia.CAM(complementary and alternative medicine)provides the option to elect favorable,low-risk,adjuvant and alternative therapies.Herein,we offer a widespread review of topical marketed formulations,natural products,and CAM treatment options for AGA.

Quantification of aflatoxins and health risk assessment through intake of herbal medicine formulations

Aflatoxins(AFs)are frequent contaminants of herbs,which pose a health risk to regular consumers.This study quantified AFs(AFB1,AFB2,AFG1,AFG2,and Total AFs)in 100 of frequently used herbal medicine formulations in Northern Jordan,and assessed the health risk associated with its intake.About half of the samples had levels lower than the detection limit,while the concentrations of AFB1 and Total AFs were higher than the European limits in 10%and 23%of the collected samples,respectively.The mean average daily dose(ADD)of AFB1 through the consumption of the formulations was 3.32×10^(-6)and ranged between 1.21×10^(-9)and 0.00045 ng/kg bwt/day,while the mean ADD of Total AFs was 6.80×10^(-5)and ranged between 3.21×10^(-9)and 0.0015 ng/kg bwt/day.The margin of exposure(MOE)for each medicine herein was far greater than 10,000 as recommended by European Food Safety Authority(EFSA)for carcinogenic and genotoxic substances.Estimated cancer risk due to life time exposure to AFB1 in herbal medicines was 1.03×10^(-5)(ranged between 4.51×10^(-4)to 1.19×10^(-9)cancers/10^(5) individuals/75 year).The frequent occurrence of AFs in the analyzed samples raised questions on their potential harmful consequences on heavy consumers,and suggested that further official measures could minimize contamination and improve management of the associated cancer risk.

Clean-DM1, a Korean Polyherbal Formula, Improves High Fat Diet-Induced Diabetic Symptoms in Mice by Regulating IRS/PI3K/AKT and AMPK Expressionsin Pancreas and Liver Tissues

Objective:To investigate the effects of Clean-DM1(C-DM1),a polyherbal formulation of Radix Scrophulariae,Radix Astragali,Rhizoma Atractylodis,and Radix Salviae Miltiorrhizae,on high-fat diet(HFD)-induced diabetes mice.Methods:The information about active components of C-DM1 extract and molecular mechanism was obtained from network pharmacology analysis.Main compounds of C-DM1 extract by high performance liquid chromatography-mass spectrometry(HPLC-MS)analysis were conducted for quality control.For in vivo study,mice were induced diabetes by HFD for 12 weeks.The mice in the normal group(Nor)were maintained with a regular diet and treated with saline by gavage.The HFD model mice were randomly divided into 3 groups,including a HFD diabetic model group,a C-DM1 extract-administered group(C-DM1,500 mg/kg),and metformin-administered groups(Met,500 mg/kg),8 mice in each group.Food intake,body weight(BW),and fasting blood glucose(FBG)levels were recorded weekly for 4 weeks.After 4 weeks of treatment,alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood glucose,low-density lipoprotein cholesterol(LDL-C)were determined using an automated clinical chemistry analyzer,and homeostatic model for assessing insulin resistance(HOMA-IR)levels and oral glucose tolerance test(OGTT)were detected.The histopathological changes of liver and pancreatic tissues were observed by hematoxylin-eosin staining.Insulin receptor substrate(IRS)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)and adenosine 5'-monophosphate-activated protein kinase(AMPK)expressions in liver and pancreas tissues were detected by Western blot analysis.Results:HPLC-MS identified dihydroisotanshinone,dihydroisotanshinone I,cryptotanshinone,harpagoside,and atractyloside A in C-DM1 extract.The administration of C-DM1 extract significantly decreased body weight,calorie intake,and the levels of blood glucose and insulin in the diabetic mice(P<0.05 or P<0.01).The C-DM1 extract administration improved the impaired glucose tolerance and insulin resistance in the diabetic mice and significantly decreased the levels of LDL-C,ALT and AST(P<0.01).The C-DM1 extract inhibited the histopathological changes of fatty liver and hyperplasia of pancreatic islets in the diabetic mice.The C-DM1 extract significantly increased the phosphorylation of IRS,AKT,and AMPK and the expression of PI3K in pancreas and liver tissues(P<0.05 or P<0.01),which was consistent with the analysis results of network pharmacology.Conclusion:C-DM1 extract improved diabetes symptoms in longterm HFD-induced mice by regulation of IRS/PI3K/AKT and AMPK expressions in pancreas and liver tissues,suggesting that C-DM1 formulation may help prevent the progression of T2DM.

Ethnomedicinal Investigation of Medicinal Plants of Chakrata Region(Uttarakhand)Used in the Traditional Medicine for Diabetes by Jaunsari Tribe

The Himalayan region is the treasure house of natural wealth,particularly of medicinal and aromatic plants.These plants are used by the Indian traditional healers for the past many centuries to treat various ailments such as skin disorders,asthma,diabetes,snake bite,fever,pain,eye diseases,diarrhoea,indigestion,jaundice,burn,wound,liver disorder,CNS disorders and urinary tract infection.The indigenous traditional knowledge of medicinal plants and therapies of various local communities has been lost due to changes in traditional culture and the introduction of modern technologies.Therefore,it is essential to explore the traditional knowledge of the indigenous medicinal plants mainly in such areas where there is a severe threat to natural vegetation owing to human inhabitation.The present study aimed to explore the medicinal plants of Chakrata region(Jaunsar-Bawar Hills),Uttarakhand,India used in the folk medicine for the management of diabetes by Jaunsari Tribe.In a comprehensive feld survey,the information about the medicinal plants have been mainly collected from the traditional healers and other elderly people belong to the tribal community.All the information about the medicinal plants of the study area was documented in a feld book.Various tools have been used to collect the samples for identifcation purpose and the authentication of the plants was done with the help of taxonomists.The literature on these plants was also searched from online(PubMed and Scopus)as well as from some textbooks and Ayurvedic classical texts.The present survey-based work described a total of 54 plants belonging to 47 genera and 30 families used in the traditional medicine for the management of diabetes in Chakrata region.The information gathered from the local community revealed that the plants are efective in diabetes and one can use most of them without consulting a practitioner or traditional healer.The literature revealed that most of the surveyed plants are already used in the preparation of various antidiabetic formulations such as Chandraprabha vati,Nishamalaki chunra,Amritamehari churna and Nisakathakadi kashayam along with various patent drugs which are frequently prescribed by the Ayurvedic practitioners in India.The present study explored the traditional as well as scientifc knowledge on the antidiabetic plants used by the tribal community.The documented information on these plants can be further used by the scientifc community to develop new drugs/formulations with the help of modern techniques.

Standardization and Quality control parameters of Kayam Churna

Kayam Churna is proprietary ayurvedic formulation which is blend of seven ingredients which are Senna leaves(Cassia angustifolia),Black Salt,Mulethi(Glycyrrhiza glabra),Nishoth(Operculina turpethum),Ajwain(Trachyspermum ammi),Haritaki(Terminalia chebula)and Svarjikshara(Sodium bicarbonate)with Senna as a major ingredient.Standardization and stability studies play a vital role for each herbal formulation,so the research was carried out to prepare the formulation in the laboratory and to standardize the Churna,by means of its physicochemical properties,phytochemical analysis and microbiology study.The organoleptic and powdered characteristics were studied and compared with monographs.Various physicochemical analysis was carried out like bulk density,tapped density,percent compressibility,loss on drying,ash value and heavy metal determination.Phytochemical analysis was carried out to find the presence of tannins,alkaloids,glycosides,flavonoids and steroids.Microbiological analysis was evaluated using the Total Viable Count method.HPTLC Chromatographic technique was used for fingerprinting.Microscopic analysis of Formulation showed the presence of Vessels,Fibre,Calcium oxalate crystals,etc.Tannins,Alkaloids,flavonoids were found to be present in the formulation.Microbial count and all physicochemical parameters were found to be within the limits as per the WHO guidelines.Further studies can be carried out for finding out its efficacy and potency.

Potential use of Clerodendrum capitatum extracts and its formulation for control of three major stored product beetles

Objective:Insecticidal activity of Clerodendrum capitatum extracts against the stored product insect pest,Sitophilus oryzae(L.),Rhyzopertha dominica(F.),and Tribolium castaneum(Herbst)were investigated.Materials and Methods:Hundred grams of C.capitatum powder was sequentially extracted with a series of solvents of increasing polarity in a Soxhlet apparatus and the extracts were concentrated on rotary evaporator to make it solvent free and the residue dissolved in a known volume of methanol and the solution was assayed for insecticidal activity by the fumigant toxicity.Hexane extract which showed maximum activity was selected for the preparation of herbal formulation and for its insecticidal properties and phytotoxicity.The herbal formulation obtained from leaves of C.capitatum was investigated by GCMS and GC/FID.Results:Clerodendrum capitatum ethyl extract had a very strong repellent activity to adults of all three major stored grain insects and was significantly repellent at a 1.58 mg/cm^(2).The hexane extract(oil)had high fumigant activity with LC_(50) values in the range between 25 and 32μg/L.A total of 14 components representing 97.8%of the formulation were identified and the main compounds in the formulation were found to be 2-Heptanone,3-methyl(33.9%)and Hexahydrofarnesyl acetone(25.3%)followed by 3-Dodecen-1-al(10.2%)and 8-Methyl-1-undecene(4.9%).Further,herbal formulation was found to be highly toxic to mixed age cultures of all three major stored grain insect pests which recorded 80%-90%mortality at dose of 200μg/L in 24 h exposure,where as 100%mortality achieved at dose of 200μg/L in 72 h exposure,respectively.Conclusion:The results indicate that extracts and formulation of C.capitatum has the potential to be developed in to a natural insecticide or repellent for controlling of stored product beetles.