Add the following:
Echinacea purpureaAerial Parts
»Echinacea purpureaAerial Parts consists of the aerial parts of Echinacea purpurea(L.)Moench (Fam.Asteraceae).It is harvested during the flowering stage.It contains not less than 1.0percent of chicoric acid,and not less than 0.01percent of dodecatetraenoic acid isobutylamides (C16H25NO)on the dried basis.
Packaging and storage— Store in tight,light-resistant containers at controlled room temperature.
Labeling— The label states the Latin binomial and,following the official name,the parts of the plant contained in the article.
USP Reference standards á11ñ— USP Chlorogenic Acid RS.USP Powdered Echinacea purpurea Extract RS.USP2E,4E-Hexadienoic Acid Isobutylamide RS.
Botanical characteristics—
MACROSCOPIC— The herb is an erect,coarse,rough-hairy perennial,usually up to 90cm tall,rarely up to 180cm.The leaves are alternate and simple;the lowermost leaves are slender,long,and petioled,ovate to broadly lanceolate,mostly penta-nerved,acute or acuminated at the apex,abruptly narrowed or rarely cordate at the base,usually sharply dentate,and 7to 20cm long and 2.5to 7.5cm wide;the petioles are mostly winged at the summit.The upper leaves are narrower,often almost entirely sessile,lanceolate or ovate lanceolate,and usually with 3veins.
The flower heads are radiate,to 15cm across,solitary or few,and long-peduncled,with 12to 20rays,purple,crimson,or rarely pale;the bristle disks are often orange,3.5to 7.5cm long;the involucre is depressed-hemispheric;the bracts are lanceolate,spreading or appressed,imbricated in 2to 4series,and hairy on the outer surface with ciliate margins;the receptacle is conical,the scales of the receptacle stiff,spinescent,and conspicuously longer than the disc flowers;the chaff is carinate and cuspidate;the achenes are 3to 4mm in length,tetrasided,obypyramidal,and thick;the pappus has a short,dentate crown.
MICROSCOPIC—
Leaf— The leaf has a thickness of 200to 350µm,with an epidermis 9to 13µm thick,largely without chloroplasts;the stomata are 28to 35µm,abundant on the dorsal surface and fewer on the ventral surface;the mesophyll is clearly divided into palisade parenchyma and sponge parenchyma.The palisade parenchyma is one layer thick,with elongated cells 50to 65µm in length,oriented at right angles to the leaf surface,containing numerous chloroplasts.The sponge parenchyma is 150to 250µm thick,with cells of irregular shape,and has multiple cell layers,few chloroplasts,and large intercellular spaces.The phloem bundles of the lateral veins within the sponge parenchyma are bound by a one-layer sheath of small parenchymous cells,with vascular elements of the midrib surrounded by large-celled parenchyma.The uniseriate trichomes are few in the ventral surface,numerous on the dorsal surface,typically tricelled,occasionally tetra-or pentacelled,250to 500µm in length,each arising from an epidermal cell;the epidermal cell walls appear with moderate thickening;the vessels are various,scalariform,with variable reticulated width.
Petiole— The parenchyma appear without chloroplasts,in several layers adjacent to a layer of collenchyma;5to 7phloem bundles of small-to medium-sized vessels are weakly lignified and embedded in the parenchyma in the form of an arc;the wing ribs of the upper surface of the slightly hollowed petiole are marginal.
Inflorescence— The epidermal cells of the ray florets are square,50µm,with a transparent,beaded cell wall;various elements of the asteraceous exhibit inflorescence;numerous multicellular jointed trichomes of the involucral bracts are 500to 800µm in length;tangential sections of the paleae with numerous fiber bundles are 10to 15µm in diameter and 100to 150µm in length;cell walls are thin.The epidermis of ray florets is reddish to violet;the epidermal cells from the end of the corolla form rounded papillae;a stigma of papillary cells is present;asteraceous pollen grains are 20to 30µm and spherical with a warty exine.
Calcium oxalate is negative;crystals of inulin and starch granules are rare.
Identification—
A: Thin-Layer Chromatographic Identification Test á201ñ—
PRESENCE OF CHICORIC ACID AND ABSENCE OF ECHINACOSIDE—
Test solution— Add 5mLof diluted alcohol (7:3)to 0.5g of the powdered plant material,and shake for 1minute.Centrifuge,and use the supernatant.
Standard solution— Dissolve an accurately weighed quantity of USP Powdered Echinacea purpurea Extract RSin methanol to obtain a solution having a concentration of about 10mg per mL.
Developing solvent system,Spray reagent 1,and Spray reagent 2— Prepare as directed for Identification test Aunder Echinacea angustifolia.
Procedure— Proceed as directed in the chapter.Develop the chromatograms in Developing solvent systemuntil the solvent front has moved not less than 18cm,and dry the plate in a stream of air.Spray the plate with Spray reagent 1followed by Spray reagent 2,and examine the plate under UVlight at 365nm:the chromatogram obtained from the Test solutionshows a yellowish-green zone at an RFvalue of 0.75due to chicoric acid and another yellowish-green zone at an RFvalue of 0.45due to caftaric acid,both zones corresponding in color and RFvalue to zones in the chromatogram obtained from the Standard solution.The chromatogram obtained from the Test solutiondoes not show or shows only traces of a zone at an RFvalue of 0.1due to echinacoside (present in Echinacea angustifoliaand in Echinacea pallida).Other colored zones of varying intensities may be observed in the chromatogram obtained from the Test solution.
B: The retention times for the relevant peaks in the chromatogram of the Test solution,mainly due to caftaric acid and chicoric acid,correspond to those in the chromatogram of Standard solution 1,as obtained in the test for Content of chicoric acid and caftaric acid.Apeak for echinacoside is not detected or is very weak.
C: The retention times for the relevant peaks in the chromatogram of the Test solution,mainly due to dodecatetraenoic isobutyl amides,correspond to those in the chromatogram of Standard solution 1,as obtained in the test for Content of dodecatetraenoic isobutylamides.
Microbial enumeration á2021ñ— It meets the requirements of the tests for absence of Salmonella speciesand Escherichia coli.The total aerobic microbial count does not exceed 105cfu per g,the total combined molds and yeasts count does not exceed 1000cfu per g,and the enterobacterial count does not exceed 1000cfu.
Loss on drying á731ñ— Dry 1g of the powdered plant material:it loses not more than 12%of its weight.
Foreign organic matter á561ñ: not more than 3.0%.
Total ash á561ñ: not more than 10.0%,determined on 3g.
Acid-insoluble ash á561ñ: not more than 2.5%.
Pesticides residues á561ñ: meet the requirements.
Heavy metals,Method IIIá231ñ: not more than 10µg per g.
Content of chicoric acid and caftaric acid—
Solvent,Solution A,Solution B,Mobile phase,and Standard solution 2— Proceed as directed in the test for Content of total phenolsunder Echinacea angustifolia.
Standard solution 1— Dissolve an accurately weighed quantity of USP Powdered Echinacea purpureaExtract RSin Solvent,shaking for 1minute,and dilute with Solventto obtain a solution having a known concentration of about 5mg per mL.Pass through a membrane filter having a 0.45-µm or finer porosity.
Test solution— Proceed as directed for Content of phenols under Echinacea angustifolia,except to use finely powdered Echinacea purpurea Aerial Parts instead of Echinacea angustifolia.
Chromatographic system (see Chromatography á621ñChromatography á621ñ)— The liquid chromatograph is equipped with a 330-nm detector and a 4.6-mm ×25-cm column that contains 5-µm packing L1.The column temperature is maintained at 35.The flow rate is about 1.5mLper minute.The chromatograph is programmed as follows.
Time (minutes) Solution A
(%)		 Solution B(%) Elution
0–13 90®78 10®22 linear gradient
13–14 78®60 22®40 linear gradient
14–17.5 60 40 isocratic
17.5–18 60®90 40®10 linear gradient
18–30 90 10 equilibration
Chromatograph Standard solution 1,and record the peak responses as directed for Procedure:the chromatogram obtained is similar to the Reference Chromatogram for total phenols provided with USP Powdered Echinacea purpureaExtract RS.Chromatograph Standard solution 2,and record the responses as directed for Procedure:the relative standard deviation for replicate injections is not more than 2%.
Procedure— Proceed as directed in the test for Content of total phenolsunder Echinacea angustifolia.Separately calculate the percentages of caftaric acid (C13H12O9)and chicoric acid (C22H18O12)in the portion of Echinacea purpureaAerial Parts taken by the formula:
2500F(C/W)(rU/rS),
in which Fis the response factor and is equal to 0.695for chicoric acid,0.881for caftaric acid,and 1.000for chlorogenic acid;Cis the concentration,in mg per mL,of USP Chlorogenic Acid RSin Standard solution 2;Wis the weight,in mg,of Echinacea purpureaAerial Parts taken;and rUand rSare the peak responses for the relevant analyte obtained from the Test solutionand Standard solution 2,respectively.Calculate the percentage of total phenols in the portion of Echinacea purpureaAerial Parts taken by adding the individual percentages calculated.
Content of dodecatetraenoic acid isobutylamides—
Mobile phase and Standard solution 2— Proceed as directed in the test for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia.
Standard solution 1— Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia,except to use USP Powdered Echinacea purpureaExtract RSinstead of USP Powdered Echinacea angustifolia Extract RS.
Test solution— Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia,except to use Echinacea purpureaAerial Parts instead of Echinacea angustifolia.
Chromatographic system (see Chromatography á621ñ)— Proceed as directed for Content of dodecatetraenoic acid isobutylamides under Echinacea angustifolia,except to use the Reference Chromatogram for alkamides provided with USP Powdered Echinacea purpureaExtract RSinstead of the Reference Chromatogram provided with USP Powdered Echinacea angustifoliaExtract RS.
Procedure— Proceed as directed in the test for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia.Identify the peaks of the two isomers of dodecatetraenoic acid isobutylamides in the chromatogram obtained from the Test solutionby comparison with the chromatogram obtained from Standard solution 1.Calculate the percentage of dodecatetraenoic acid isobutylamides in the portion of Echinacea purpureaAerial Parts taken by the formula:
10(1.353)(C/W)(rU/rS),
in which 1.353is the response factor for 2E,4E-hexadienoic acid isobutylamide;Cis the concentration,in mg per mL,of USP2E,4E-Hexadienoic Acid Isobutylamide RSin Standard solution 2;Wis the weight,in g,of Echinacea purpureaAerial Parts taken;rUis the sum of the peak responses of the relevant analytes obtained from the Test solution;and rSis the peak response obtained from Standard solution 2.USP28
Auxiliary Information— Staff Liaison:Gabriel I.Giancaspro,Ph.D.,Senior Scientist and Latin American Specialist
Expert Committee:(DSB)Dietary Supplements:Botanicals
USP28–NF23Page 2080
Pharmacopeial Forum:Volume No.30(2)Page 557
Phone Number:1-301-816-8343