Marketplace Analysis Demonstrates Quality Control .
Plant Foods Hum Nutr (2014) 69:161–167DOI 10.1007/s11130-014-0416-yORIGINAL PAPERMarketplace Analysis Demonstrates Quality Control StandardsNeeded for Black Raspberry Dietary SupplementsJungmin LeePublished online: 25 April 2014# The Author(s) 2014. This article is published with open access at Springerlink.comAbstract There is currently no standard for the minimumanthocyanin concentration a black raspberry dietary supplement must contain for legal sale in the US. All consumeravailable black raspberry products (n 19), packaged as dietary supplements or otherwise prepared (freeze-dried wholeand pre-ground powders), were purchased and analyzed fortheir anthocyanin composition and concentration. Seven ofthe 19 samples contained no anthocyanins from black raspberry fruit, while three of those seven (without black raspberryfruit) had no anthocyanins of any kind. There was a widerange of anthocyanin concentration within the remainingproducts (18.1–2,904.8 mg/100 g; n 12). When expressedas per capsule or per 1 teaspoon, concentration ranged from0.1 to 145.2 mg (average 28 mg; n 12). Until US dietarysupplement labeling comes under regulatory oversight similarto food guidelines, foods are a more dependable source fordietary phenolics than supplements.Keywords Authenticity . Adulteration . Pigment . Rubus .Rubus occidentalis L . Black capIntroductionThe USDA (United States Department of Agriculture) has puttogether several campaigns advocating a healthy eating dietby having one that is color diverse (USDA 5 A Day campaign,USDA ChooseMyPlate.gov, HealthierUS School ChallengeJ. Lee (*)United States Department of Agriculture, Agricultural ResearchService, Horticultural Crops Research Unit Worksite,29603 U of I Ln., Parma, ID 83660, USAe-mail: jungmin.lee@ars.usda.govJ. Leee-mail: jlee@uidaho.eduHUSSC, etc.). As consumers have become increasingly awareof the benefits from eating healthier, sales of black raspberrysupplements have also increased, and were further raised aftera popular US media program promoted black raspberry consumption for its cancer fighting potential. This popular television show made the assertion that an adult should consume600 mg of anthocyanins from black raspberry dietary supplements daily (300 mg twice a day), followed by the unreasonable claim that each 300 mg capsule (featured product)contained the equivalent content of four cups of fresh blackraspberries. Two samples linked with the group making thesestatements were examined in this study, and are discussedlater. Although the potential health benefits of black raspberryfruit and its specific mechanisms are still under investigation[1–3], unwitting consumer demand has increased the availability of products described as black raspberry supplementsin the dietary supplement marketplace.Black raspberry (Rubus occidentalis L.; native to EasternNorth America) fruit has been traditionally used as a food anda natural colorant, but renewed US consumer interest hasbrought an upsurge in the number of commercial black raspberry products available (from desserts to dietary supplements; [4]). This can partially be explained by increasedawareness of the potential health benefits high-pigmented fruitmight provide [1–3, 5, 6], but their distinct flavor, unlikeblackberries or red raspberries, may also help. Unfortunateside effects of intensified consumer demand have been occurrences of product adulteration, though some likely arose fromthe limited production of black raspberries, which due to theirunique growing requirements make Oregon the only US statewith notable acreage [4].Black raspberry (Rubus occidentalis L.) has fruit, plantphenology, plant morphology, and anthocyanin profile distinctly different from red raspberry, blackberry, or any othergenus Rubus berries [5, 7–9]. Since most people are unable todistinguish black raspberries, a simple fact sheet has been
162generated (www.black-raspberry.com) to help educateresearchers, health professionals, industry, and consumersthe differences among black raspberry, red raspberry, andblackberry. It is apparent that black raspberry dietarysupplement producers and sellers might not be able todifferentiate between black raspberry and blackberry, as theimages on the supplements’ packaging were incorrect in fourof the products examined in this study (summarized inTable 1). The inaccurate images used were that of eitherblackberries or an altered image of blackberries (white corewas colored in black). The confusion about black raspberryfruit’s correct morphology [8, 9], and the sudden boost inavailable black raspberry products (although the fruitproduction is limited; [10]), caused us to examine theanthocyanin concentration of marketplace black raspberrydietary supplements, and other forms that can be used assupplements (i.e., freeze-dried black raspberries, extracts).Materials and MethodsSamples, Reagents, Chemical, and Standards An effort topurchase all commercially available black raspberry supplements and dried fruit (powder and whole fruit forms) weremade (n 19) from May to July of 2013 (Amazon.com, Inc.,Seattle, WA, USA). No purchased products were past theirexpiration or best use by date. These samples representedproducts from 17 companies. Products A09, B01, and B02were from one company. The rest of the samples were fromdifferent companies. Sample information is summarized inTable 1. Dietary supplements in capsules or extract werecoded A01 to A15, with one sample (A09) in liquid form.The four available dried fruit products were purchased andcoded B01 to B04. One sample was freeze-dried whole fruit(B01); while the remaining dried samples were in powderform. Five capsules and their contents were weighed in triplicate to determine the weight of the powder within the capsules, and to convert our findings into per capsule. Capsulecontents of A11 were suspicious (see Table 1) and a secondexample was purchased to double check that the originalproduct was not random error.All chemicals, reagents, and standards used in this studywere analytical or HPLC grade from Sigma-Aldrich ChemicalCo. (St. Louis, MO, USA). Cyanidin-3-glucoside was purchased from Polyphenols Laboratories AS (Sandnes, Norway).Extraction and Sample Preparation All powder contents ofthe capsules were removed and stored at 75 C until extraction. The one example of freeze-dried whole fruit (B01) wasground (using a coffee bean grinder, model K2M2; BraunGmbH, Kronberg, Germany) prior to storage and subsequentextraction. Each sample group’s collected powders werepooled and kept frozen until the start of chemical extraction.Plant Foods Hum Nutr (2014) 69:161–167Samples were extracted and expressed as-is, since that represented the form of intended consumption. Powders (initially1.5 g) were extracted with high purity water (initially 15 mL;Millipore Simplicity UV, Millipore Corp., Billerica, MA,USA) by sonication for 15 min, centrifuged 10 min at4,000 rpm, then filtered (Millipore 0.45 μm Millex-FH syringe filter, Bedford, MA, USA) prior to injection onto theHPLC system [12, 13]. Solid to liquid extraction ratio had tobe altered for samples that indicated very low to zero levels ofanthocyanins (3.0 g powder extracted in 10 mL of water). A09was diluted (1.5 g:15 mL high purity water; by weight due toits high viscosity) and put through the same process as theother samples (sonication, centrifugation, and filtration) priorto HPLC analysis. ‘Munger’ fruit extract was obtained fromprevious study [10, 11, 14, 15].HPLC (High Performance Liquid Chromatography) Condition for Individual Anthocyanin Separation HPLC/DAD (diode array detector)/MS (mass spectrometry) was used foranthocyanin elution as described in detail in our previouswork [13], except for the use of a longer analytical column[8]. Briefly, an Agilent HPLC 1100 (Agilent TechnologiesInc., Palo Alto, CA, USA) was used for this investigation.Individual peaks were monitored at 520, 280, and 255 nm.Anthocyanins were expressed as cyanidin-3-glucoside (Polyphenols Laboratories AS, Sandnes, Norway). Anthocyaninpeaks were identified by retention time, UV–VIS spectra,external standards (when available), verified fruit with knownanthocyanin profiles, and prior published research [8, 10, 11,14, 16–18]. Analyses were conducted in duplicate. Resultswere expressed as mg of cyanidin-3-glucoside/100 g of powder, mg of cyanidin-3-glucoside/capsule for samples codedA01–A08, or 5 g ( 1 teaspoon) for samples coded A09 andB01–B03. Peaks 2 and 3 peak areas were split at each apex,where it was not co-eluting, and multiplied by two to obtaintotal peak area prior to calculations.Results and DiscussionRelevant information from packaging, prices, and observations are summarized in Table 1; including the labeled contents from the products that were found to contain no blackraspberry fruit. Photo examples of six dietary supplements canbe found in Fig. 1. Supplements ranged in visual color fromlight pink to dark red, except for A11 capsules (see Fig. 1.e)that contained no red hue. An example of no filler added in thecapsule contents (Fig. 1.a) compared to high amounts of fillerused in Fig. 1.b through Fig. 1.d. Red appearance cannot beused to indicate the presence of black raspberry fruit sincesome of the manufacturers used pink colored fillers (A10 andA12) and other fruit powder (A12, A14, A15, and B04).
Plant Foods Hum Nutr (2014) 69:161–167163Table 1 Sample codes, brief summary of label information (codes used in Table 2), and observations. All samples were labeled on the bottle or packagethat contained black raspberrySample Information provided on product packaging. Cost per capsulecodeor teaspoon. Entire package price.Observations and comments.A01500 mg of black raspberry fruit extract. 0.11/capsule. Total cost 6.30. Misspelled Rubus leucodermis on bottle. Latin name used on label maybe incorrect, as the majority of commercially grown black raspberriesare R. occidentalis not R. leucodermis [10]. Contained unlisted nonfruit ingredients. Capsule content weight was 0.49 g.A02One capsule contains 425 mg of R. occidentalis berry. 0.17/capsule.Total cost 19.95.550 mg Rubus occidentalis berry. 0.13/capsule. Total cost 11.99.A03Contained unlisted ingredients. Capsule content weight was 0.81 g.Label misspelled ‘berry’. Contained unlisted ingredients besides driedfruit powder. Label had correct black raspberry image. Capsulecontent weight was 0.48 g.A04100 % pure black raspberry dietary supplement. 400 mg of blackraspberry powder per capsule. Vegetable capsule (plant derivedcellulose). 0.31/capsule. Total cost 18.39.Capsule content did not appear to be pure black raspberry fruit powder.Capsule content weight was 0.46 g. Contained unlisted non-fruitingredients.A05One capsule contains 250 mg of black raspberry. 0.15/capsule. Totalcost 18.34.Blackberry image on label. Contained unlisted non-fruit ingredientsbesides dried fruit powder. Capsule content weight was 0.61 g.A06300 mg of black raspberry (as Rubus occidentalis berry). 0.18/capsule. Contained unlisted non-fruit ingredients besides dried fruit powder.Total cost 16.05.Capsule content weight was 0.30 g.A07300 mg of Rubus occidentalis fruit. Freeze-dried. 0.15/capsule. Totalcost 8.99.A08Each capsule contains 300 mg of black raspberry (Rubus occidentalis). Capsule content weight was 0.32 g. Capsule contents appeared to beFreeze-dried fruit. 0.14/capsule. Total cost 12.95.pure black raspberry fruit powder.A09Clinical strength black raspberry extract. Contains 6 % alcohol.Anthocyanin equivalent to 500 fresh black raspberries or 100 g offreeze-dried powder. Fruit grown in Oregon. 2.50/teaspoon. Totalcost 29.99.A10One 425 mg capsule has the micronutrient equivalent of over 4 cups of Blackberry image on label. No 520 nm absorbing compounds.fresh berries. 0.11/capsule. Total cost 6.50.Contained unlisted non-fruit ingredients besides dried fruit powder.May have included colored filler. Capsule content weight was 0.54 g.A11Fresh, raw, pure. 400 mg of seedless black raspberry powder. Vegetarian Modified blackberry image on label with the white core was blacked out.Dark olive-brown-black powder in capsule did not look like berrycapsules and absolutely nothing else. 0.79/capsule. Total costpowder and had a medicinal odor. No 520 nm absorbing compounds. 23.77.This sample contained no black raspberry fruit. Capsule contentweight was 0.41 g. See photo image in Fig. 1.e.A12425 mg of black raspberry fruit. 0.18/capsule. Total cost 10.95.Contained 520 nm absorbing compounds, but not black raspberryanthocyanins. Contained unlisted non-fruit ingredients. May haveincluded colored filler. Capsule content weight was 0.51 g.A13425 mg Rubus occidentalis fruit. 0.06/capsule. Total cost 3.13.No 520 nm absorbing compounds. Contained unlisted non-fruitingredients besides. Capsule content weight was 0.47 g.A14300 mg black raspberry. Made in USA. 0.18/capsule. Total cost 15.95.Contained 520 nm absorbing compounds, but not black raspberryanthocyanins. Only sample in opaque capsules. Contained unlistednon-fruit ingredients. Capsule content weight was 0.57 g.A15100 % vegetarian, black raspberry, pure Rubus occidentalis extract.500 mg of extract per capsule. 0.23/capsule. Total cost 23.14.Contained 520 nm absorbing compound (very early eluting compound),but not black raspberry anthocyanins. Contained unlisted non-fruitingredients. Capsule content weight was 0.65 g.B01Freeze-dried whole fruit was in a Millard food bag and vacuumedFreeze-dried whole fruit. Vacuumed packed. USDA grade A. Nopackaged. Label had correct black raspberry image.preservatives. Grown in Oregon. 100 % pure Oregon grown blackraspberry. 1.20/teaspoon. Total cost 23.99.Powder was in a Millard food bag and vacuumed packaged. Label hadFreeze-dried powder. USDA grade A. No preservatives. Grown incorrect black raspberry image.Oregon. 100 % pure Oregon grown black raspberry. 1.43/ teaspoon.Total cost 28.63.Black raspberry powder dietary supplement. Instructions forPowder was in a packaged in a can. Label had correct black raspberrysupplementing fruit in meals. 1.02/teaspoon. Total cost 29.20.image.B02B03B04Live organic black raspberry. Instructions on how to consume as adietary supplement. 2.38/ teaspoon. Total cost 49.99.Blackberry image on label. Capsule content weight was 0.32 g. Capsulecontents appeared to be pure black raspberry fruit powder.Only liquid (extract) form available. Company claims clinical strengthbased on Stoner et al. [1] work. Description indicated a juiceconcentrate with small amount of alcohol. Appeared as a highlyviscous liquid, like a juice concentrate. Label had correct blackraspberry image.Contained compounds that absorb at 520 nm, but not black raspberryanthocyanins (see Fig. 2.). One side of the bag was transparent. Thispowder had a different hue than samples B02 and B03 (appeared to benot as intense red-purple).
164Plant Foods Hum Nutr (2014) 69:161–167Fig. 1 Photos of six blackraspberry dietary supplementspurchased. In photo example bthrough d, contained highamounts of filler. In photoexample e (coded A11 in Table 1),contained no anthocyanin (noblack raspberry fruit)Seven (A10–A15 and B04) of the 19 samples had no blackraspberry anthocyanins. Out of those seven, A10, A11, andA13 contained no 520 nm absorbing compounds (chromatogram of A10 as an example in Fig. 2). Samples A12, A14,A15, and B04 contained 520 nm absorbing peaks, but theirprofiles did not match that of black raspberry (examples inFig. 2.a and b). One anthocyanin profile, B04 from fourfreeze-dried samples (which were the B coded samples), didFig. 2 Anthocyanin profiles of black raspberries (a- ‘Munger’ and bA09) and selected supplement samples (c- A10, d- A12, e- A14, and fB04) that contained questionable materials other than black raspberryfruit. The chromatograms were monitored at 520 nm (280 and 255 nmtraces not shown). Clearly, black raspberry anthocyanin profile is suitablefor authenticity work (comparing ‘Munger’ to sample A09, leaves nodoubt that A09 contains black raspberry). Corresponding peak identification for ‘Munger’ and A09 listed in Table 2. Additional black raspberryanthocyanin profiles can be found in our past work [7, 8, 10, 11]
Plant Foods Hum Nutr (2014) 69:161–167165not match black raspberry either (see Fig. 2). We suspect B04was blackberry (Rubus spp.; [5, 19]) freeze-dried powder andsold as black raspberry. Hydrochloric acid was used to confirm that A10, A11, and A13 contained no anthocyanins, as acolor shift (redness due to oxonium formation) after acidification would also indicate the presence of anthocyanin[20–22], but this visual change did not occur in these threeextracts. Some example chromatograms of these questionablematerials are shown in Fig. 2 (c through f). Our secondpurchased A11 sample had an identical appearance (olivebrown-black powder; see Fig. 1.e) and the same HPLC profile(data not shown) as the first A11 sample, again with nodetectable 520 nm absorbing peaks. Sample A03’s labelclaimed 550 mg of black raspberry in each capsule, thoughcapsule entire content’s weight was measured at 480 mg(Table 1).Cyanidin-3-rutinoside was the main anthocyanin amongthe twelve samples that did contain black raspberry fruit(Table 2). Six samples (A07, A08, A09, B01, B02, and B03)contained all seven anthocyanins routinely found in blackraspberry fruit [5, 11, 14, 15]. These seven black raspberryanthocyanins in the order of elution are cyanidin-3sambubioside, cyanidin-3-xylosylrutinoside, cyanidin-3glucoside, cyanidin-3-rutinoside, , and peonidin-3-rutinoside. Sixsamples (A01 through A06) contained five of the sevenanthocyanins, with only pelargonidin-3-glucoside andpeonidin-3-rutinoside not being detected. Representativeblack raspberry fruit anthocyanin chromatograms can befound in Fig. 2 (a and b).Black raspberry labeled products (n 19; Tables 1 and 2)contained zero to 2,904.8 mg/100 g of anthocyanin. Productsthat did contain black raspberry fruit anthocyanins ranged inconcentration from 18.1 to 2,904.8 mg/100 g (n 12; 160fold difference), and 0.1 to 145.2 mg/capsule or 1 teaspoon(n 12; 1,400 fold difference). The wide range in anthocyanins found in the dietary supplements are not surprising sinceour past work observed the fruit itself to vary from 3 to996 mg/100 mL of fresh fruit (n 1,000; [4, 10, 11, 14, 15]).But, a partial explanation for the wide ranges observed herewere due to non-fruit ingredients (fillers, binders, bulkingagents, carriers, etc.) such as rice powder, silica, magnesiumstearate, etc. The black raspberry fruit containing capsules(A01–A08) averaged 305.5 mg/100 g or 1.0 mg/capsule,while A09 (the only sample in extract form) contained2,904.8 mg/100 g or 145.2 mg/5 g ( 1 teaspoon). The blackraspberry fruit containing freeze-dried products (B01–B03)averaged 1,217.9 mg/100 g or 60.9 mg/5 g ( 1 teaspoon).All samples were lower in anthocyanin than what had beenpreviously reported for freeze-dried black raspberry powder:3,200 mg/100 g (cultivar Jewel; [1]) and 4,360 mg/100 g(unknown cultivar; [23]). Possible contributors to these discrepancies include differences in sample handling (freezedrying preparation, extraction, storage, etc.) and analysis conditions [5, 7].A07 and A08 appeared to be pure fruit powders as indicated by their uniform dark red-purple powders (whole blackraspberry seeds present) and high anthocyanin levels (A071052.4 and A08- 1,138.8 mg/100 g). Based on observing A07and A08, it would be possible to fill capsules without bulkingTable 2 Individual anthocyanin concentrations are listed in the order ofHPLC elution (peak 1: cyanidin-3-sambubioside, peak 2: cyanidin-3xylosylrutinoside, peak 3: cyanidin-3-glucoside, peak 4: cyanidin-3-rutinoside, peak 5: pelargonidin-3-glucoside, peak 6: pelargonidin-3rutinoside, and peak 7: peonidin-3-rutinoside)Sample codePeak 6Peak 1Peak 2Peak 3Peak 4Peak 5Peak 7Total (mg/100 8.1 (0.1)21.6 7.413.413.221.8ndndndnd3.13.67.32.62.76.525.9 (0.1)35.7 (0.2)36.8 (0.2)114.8 (0.3)1052.4 (3.4)1138.8 (3.6)2904.8 (38.5)828.5 (37.0)1082.8 (23.3)1742.2 (85.9)Totala (mg/capsule or 5 g)0.1 (0)0.2 (0.1)0.1 (0)0.2 (0)0.2 (0)0.3 (0)3.4 (0.1)3.6 (0.1)145.2 (1.9)41.4 (1.8)54.1 (1.2)87.1 (4.3)Total expressed as mg of cyanidin-3-glucoside/100 g and mg/capsule or 5 g ( 1 teaspoon). Values within parenthesis indicate standard errors. Sevensamples (A10, A11, A12, A13, A14, A15, and B04) either contain no detectable anthocyanins or no black raspberry anthocyanins (see Table 1)aA01–A08 were expressed as per capsule, A09 and B01–B03 as per 1 teaspoon (5 g)bnd, not detected
166agents and fillers as seen in the lower quality capsule samples.The highest anthocyanin concentration was from A09 (theliquid example) at 2,904.8 mg/100 g. If one consumed twocapsules from sample A08 or A09, they ingested only 7.0 mg of anthocyanins. Samples B01, B02, and B03 ifpackaged as capsules, would provide amounts similar toA07 and A08, since those two capsule products contained100 % freeze-dried berries.Samples A04 and A11 are linked to the unnamed groupmentioned in the introduction. A04 contained 0.2 mg/capsule,while A11 contained the mysterious olive-brown-black powder with a strange medicinal odor. A04 contained ingredientsother than fruit powder that were not listed on the label(Table 1). In fact, three samples (A04, A06, and A11) didnot indicate additional ingredients in capsules, but based onthe visual appearance and HPLC anthocyanin results it is clearthey did contain substances other than black raspberry fruitpowder.Using anthocyanin profile for identifying food and dietarysupplement adulteration is not a new concept and has beendemonstrated before [7, 8, 24, 25]. Misidentification of plantsource material is a known issue in the US dietary/herbalsupplement industry [26–29] and it is an obvious problemthat needs to be corrected. It is possible that due to supplydemand that the dietary supplement companies were fraudulent, or made honest mistakes from not testing constituentsprior to production. Some issues surrounding dietary supplements can be resolved by improved dietary supplement regulations, and by endorsing proposed rules [30–34] for the safetyof US consumers before another “ephedrine” scale incidentoccur [29].ConclusionWhile there are companies (four companies herein) that provide consumers with high black raspberry anthocyanin containing products (A07, A08, A09, B01, B02, and B03), themajority ( 70 %) of companies are selling low quality products, some containing unknown/unreported ingredients andvery little black raspberry fruit. Until US dietary supplementproducts are better regulated and quality control standards forsafety, purity, and dosage are defined and endorsed, the safersource for dietary phenolics as a consumer is from foodintake [6]. From past research [24, 35, 36] and findingsherein, there is a real need to create standards fordietary supplements made from plant sources. At themoment, a consumer who assumes the US dietary supplementmarketplace is free from risk is unfortunately naive. Fortypercent (seven out of 19) of the black raspberry supplementsand products purchased and evaluated here contained no blackraspberry fruit anthocyanin.Plant Foods Hum Nutr (2014) 69:161–167Acknowledgments We thank Dr. Michael Dossett of BC BlueberryCouncil (in partnership with Agriculture and Agri-Food Canada) and Dr.Chad E. Finn of USDA for ‘Munger’ fruit. This project was funded byUSDA-Agricultural Research Service (ARS) CRIS number 5358-21000047-00D and a Specialty Crop Research Initiative (SCRI) grant number2011-51181-30676 from USDA-National Institute of Food and Agriculture (NIFA). Mention of trade names or commercial products in thispublication is solely for the purpose of providing specific informationand does not imply recommendation or endorsement by the U.S. Department of Agriculture.Conflict of Interest NoneOpen Access This article is distributed under the terms of the CreativeCommons Attribution License which permits any use, distribution, andreproduction in any medium, provided the original author(s) and thesource are credited.References1. Stoner GD, Wang L, Zikri N, Chen T, Hecht SS, Huang C, Sardo C,Lechner JF (2008) Cancer prevention with freeze-dried berries andberry components. Semin Cancer Biol 17:403–4102. Stoner GD, Sardo C, Apseloff G, Mullet D, Wargo W, Pound V, SinghA, Sanders J, Aziz R, Casto B, Sun X (2005) Pharmacokinetics ofanthocyanins and ellagic acid in healthy volunteers fed freeze-dried blackraspberries daily for 7 days. J Clin Pharmacol 45:1153–15643. 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Capsule content weight was 0.30 g. A07 300 mg of Rubus occidentalis fruit. Freeze-dried. 0.15/capsule. Total cost 8.99. Blackberryimageonlabel.Capsulecontentweightwas0.32 g.Capsule contents appeared to be pure black raspberry fruit powder. A08 Each capsule contains 300 mg of black raspb
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If you're eligible for health coverage through Medicaid or the Children's Health Insurance Program (CHIP), you can't get help paying for health coverage through the Marketplace. You should do one of these: End your enrollment in Marketplace coverage Update your application to tell the Marketplace you're not enrolled in Medicaid or CHIP
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Private Student Loan Marketplace for private education loans that are made through the Marketplace process for the reasons below. We believe that the Marketplace promotes competition among the participating lenders and such competition may result in borrowers receiving better terms and conditions on private education loans.
