In collaboration with universities and specialized laboratories, Promedivet evaluates the phytochemical profile and relevant bioactive markers of selected herbal raw materials and finished formulations. These analyses support scientifically guided ingredient selection, formulation optimization, product standardization, quality control, and batch-to-batch consistency.

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Why Measure at All

A herbal formulation is not a molecule with a fixed dose; it is a plant matrix whose composition moves with the species, the harvest, the season, the solvent and the process. That is the honest starting point, and it is also the reason this work exists. Without measurement there is no way to tell a good batch from a weak one, no basis for choosing one plant over another, and nothing to put on a specification. Measurement is what turns a traditional recipe into a product that can be made the same way twice.

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Four analytical questions run through everything below: what is in the plant (raw material screening), what survives into the extract (formulation), what the finished product must contain to be released (specification), and how much that varies between batches (consistency). Each has its own data set, and they are reported separately below rather than merged into a single claim.

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Analytical Methods

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Table 1. The analytical panel applied across raw materials, developmental extracts and finished products. Two independent antioxidant chemistries are used rather than one, because DPPH and ABTS respond differently to different compound classes; agreement between them is itself a quality signal.

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All spectrophotometric determinations are run in triplicate and reported as mean ± standard deviation, with ANOVA and significance at p < 0.05. The published work correlated polyphenol and flavonoid content against both DPPH and ABTS results, and found that phenolic content does track antioxidant activity across this material - which is what justifies using the cheap, fast total-phenolic assay as a routine release test once the relationship has been established by the more expensive instrumental work.

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1. Ingredient Selection: Screening the Raw Materials

Sixteen medicinal plants from the spontaneous flora were screened before they entered any formulation, each extracted three ways - ethanol 70°, methanol:water (1:1) and methanol - so that the solvent effect could be separated from the plant effect. Nine of them were carried forward into antioxidant testing. The spread is wide and it matters: total polyphenols ranged from 3.63 to 48.70 mg GAE/100 g dry weight and flavonoids from 0.25 to 38.5 mg QE/100 g depending on species and solvent.

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Table 2. Highest-yielding solvent per species, from the sixteen-plant screen (Varga et al., 2018, Table I). Values are per 100 g dry plant weight.

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The screen also produced the two findings that shape formulation. First, the richest plant is not always the most active one: the aqueous extract of Origani herba gave the single best DPPH result of the whole plant set (IC50 1.70 µg/mL) despite a modest flavonoid figure, while Urticae, Hyperici and Origani herba led on ABTS. Second, the strongest laboratory solvents are unusable in practice - ethanol 70°, methanol and methanol:water cannot be used in registered nutritional supplements for animals, so the commercial extracts are made with propylene glycol. The screening data therefore rank the plants, not the products; the products had to be measured separately.

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2. The Finished Formulations: Profile and Antioxidant Capacity

Fifteen finished propylene-glycol tinctures were measured directly, in liquid form, for total flavonoids, total polyphenols and antioxidant capacity by both methods. This is the reference profile of the range.

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Table 3. Phytochemical and antioxidant profile of the fifteen finished tinctures. Values from Varga et al. (2018), Tables I and II, where the extracts appear anonymised as "Extract number 1–15"; product names come from the corresponding internal table, which reproduces the same figures against named products. See the note on this pairing below.

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Read across the table, the practical point is the antioxidant column. Every one of the fifteen finished products scavenges radicals at IC50 values between 0.71 and 4.40 µg/mL by DPPH and 0.01 and 2.89 µg/mL by ABTS - a narrow band at the potent end of the scale, and a much tighter clustering than the raw plant extracts showed (DPPH 1.70 to 517.22 µg/mL). The formulation process is doing work here: it is not merely diluting the plants, it is delivering a consistent level of activity across products built from different botanical bases. The strongest performers on DPPH were Herba-Top Entero for poultry (0.71) and for mammals (0.79) and Rumdigestin (0.81); the weakest was Trichomonas for mammals (4.40), still well inside the potent range.

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3. Product Standardization: From Measurement to Specification

Analysis only becomes quality control when it produces a number a batch has to hit. The range is covered by a certificate-of-analysis format that sets acceptance ranges for each measured parameter and records the batch result against them. Organoleptic checks sit alongside the instrumental ones, and seven individual marker compounds are quantified by LC/MS-MS rather than only the two summed totals.

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‍Table 4. Certificate-of-analysis format, illustrated for Coccidioza Mamifere. Spectrophotometric parameters by Folin-Ciocalteu and AlCl₃ colorimetry; individual markers by LC/MS-MS.

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Two things are worth drawing out. The ranges are genuine windows, not rubber stamps - chlorogenic acid must land between 50 and 100 µg/mL, luteolin between 2 and 12 - and a batch outside them fails. And the recorded results sit comfortably mid-range rather than hugging a limit: polyphenols at 4.58 against a 2.0–10.0 window, chlorogenic acid at 75.9 against 50–100. The totals in this certificate are also identical to the values published for the same product in the peer-reviewed screen, which means the release test and the research method are measuring the same thing the same way.

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4. Batch-to-Batch Consistency: What Actually Varies

The most useful data set here is also the least flattering, and it should stay on the page for exactly that reason. Seven veterinary products were measured twice, on material from January and from July, and the difference is systematic:

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Table 5. Seasonal comparison of polyphenol and flavonoid content in seven veterinary products. Flavonoid standard deviations in the source are given in the order of 10⁻² and are omitted here for readability.

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Every one of the seven products lost polyphenol content between January and July - between 40% and 61%, with the anti-allergy formulation showing the largest drop and the anticoccidial the smallest. Flavonoid content, by contrast, stayed essentially flat, moving only in the second decimal and in both directions. The two markers are therefore not interchangeable: the phenolic fraction is where seasonal variability shows up, and the flavonoid fraction is comparatively stable.

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This is the argument for the whole analytical programme, stated in one table. A product made from wild-harvested plants will vary if nothing is done about it; measuring each batch and releasing against a defined range is what keeps the delivered product inside a known window. It is also a practical input for production planning - harvest timing and the raw-material-to-solvent ratio are the levers that close the gap.

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Research Partners and Sources

The analytical work behind this page was carried out with the Faculty of Pharmacy, University of Medicine and Pharmacy of Tîrgu Mureș - Department of Pharmacognosy and Phytotherapy, Department of Pharmaceutical Chemistry, and Department of Toxicology and Biopharmacy - with a contribution from the Department of Pharmacognosy, Semmelweis University, Budapest. Botanical identification and voucher specimen deposition were handled in Tîrgu Mureș.

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Study 1 - Phenolic content and antioxidant screening

Publication:  Varga, E., Schmidt, I., Szövérfi, B., Pop, M.D., Kelemen, H., Tóth, A. (2018). Phenolic content from medicinal plants and their products used in veterinary medicine. Acta Medica Marisiensis 64(2), 77–82. Peer-reviewed, open access.

Scope:  16 medicinal plants × 3 solvents, plus 15 finished propylene-glycol tinctures; TPC, TFC, DPPH and ABTS. Triplicate determinations, mean ± SD, ANOVA, p < 0.05.

Funding:  Internal research grant No. 17972/07.12.2016, UMF Tîrgu Mureș and SC Promedivet SRL, and Erasmus Student Mobility for Placements (2016–2017). Declared conflict of interest: none.

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Study 2 - Marker quantification by UHPLC-MS/MS

Publication:  Varga, E., Fülöp, I., Farczádi, L., Croitoru, M.D. (2020). Polyphenolic determination from medicinal plants used in veterinary medicine by an UHPLC-LC-MS/MS method. Farmacia 68(6), 1129–1135. Peer-reviewed.

Role:  Establishes the instrumental method behind the individual marker figures used in the certificate of analysis, and supplies the quantified profile cited on the Salmonellosis page for Herba Top Salmonella (apigenin, kaempferol, quercetin, luteolin, quercitrin, chlorogenic acid, caffeic acid).

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Internal quality documents

• Buletin de analiză (certificate of analysis) format, nr. x/2019 - specification ranges and result fields per product. Template; unsigned and undated in the supplied copy.
• Seasonal polyphenol and flavonoid determination across seven veterinary products (January vs July). Source document not identified in the supplied copy.

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References

1. Varga, E., Schmidt, I., Szövérfi, B., Pop, M.D., Kelemen, H., Tóth, A. (2018). Phenolic content from medicinal plants and their products used in veterinary medicine. Acta Medica Marisiensis, 64(2), 77–82. DOI: 10.2478/amma-2018-0013.
2. Varga, E., Fülöp, I., Farczádi, L., Croitoru, M.D. (2020). Polyphenolic determination from medicinal plants used in veterinary medicine by an UHPLC-LC-MS/MS method. Farmacia, 68(6), 1129–1135. Article page.
3. Singleton, V.L., Rossi, J.A. (1965). Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144–158. (Folin-Ciocalteu reference method.)
4. Farmacopeea Română, 10th edition. Bucharest, 1993, p. 335. (Official total flavonoid method.)
5. Brand-Williams, W., Cuvelier, M.E., Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. Lebensmittel-Wissenschaft und -Technologie, 28, 25–30. (DPPH reference method.)
6. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9–10), 1231–1237. (ABTS reference method.)
7. Buletin de analiză nr. x/2019 - Coccidioza Mamifere. SC Promedivet SRL certificate-of-analysis format. (Internal document; template, unsigned and undated in the supplied copy.)
8. Flavonoid és összpolifenol tartalom meghatározása állatgyógyászati termékekből - seasonal determination across seven veterinary products, January vs July. (Internal document; parent source not identified in the supplied copy.)

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Raw Material and Supplier Transparency

• Manufacturer: SC Promedivet SRL, Sovata, Romania.
• Botanical sourcing: sixteen species grown in the spontaneous flora at Sovata, Mureș County, harvested July–August 2015 for the screening study, with voucher specimens deposited at the Department of Pharmacognosy and Phytotherapy, University of Medicine and Pharmacy of Tîrgu Mureș.
• Species screened: Agrimonia eupatoria, Arctium lappa, Artemisia absinthium, Betula pendula, Calendula officinalis, Convolvulus arvensis, Equisetum arvense, Hypericum perforatum, Lythrum salicaria, Origanum vulgare, Plantago lanceolata, Polygonum bistorta, Potentilla anserina, Symphytum officinale, Thymus serpyllum, Urtica dioica.
• Extraction: propylene glycol diluted with water (70%), by direct pressing in a hydraulic press. Propylene glycol rather than ethanol 70° because nutritional supplements for animals cannot be registered with ethanol as the solvent.

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