Verification · Certificates issued
Certificate AN-2026-00017
Valid
Issued
This certificate was issued by ANALYTON and has not been withdrawn.
Verification confirms that a certificate with this number and key was issued by ANALYTON on the date shown, for the sample described, and states its current status.
| Report number | AN-2026-00017 |
| Issued | 2026-09-15 |
| Status | Valid |
| Sample | beta-Nicotinamide adenine dinucleotide (NAD+), free acid, powder, vial labelled 500 mg |
| Batch number | ATNAD-500-2608-01 |
| Matrix | Research compound, small molecule |
| Determinations performed | 22 |
| Issued by | ANALYTON |
| Certificate file | AN-2026-00017.pdf |
01Results 22
| Determination | Method | Result | Specification |
|---|---|---|---|
| Appearance | Visual | White to off-white crystalline powder, hygroscopic, free of visible particles | White to off-white powder Pass |
| Identity | LC-HRMS, ESI+ and ESI-, USP <621> principles; UV | [M+H]+ 664.1168, deviation 0.6 ppm; [M-H]- 662.1014, deviation 0.6 ppm; MS/MS 542.069 (loss of nicotinamide, 122.048 Da), 428.037 (ADP), 348.070 (AMP), 136.062 (adenine), 123.055 (nicotinamide); UV λmax 259 nm, no absorbance at 340 nm (oxidised form) | Theoretical monoisotopic 663.1091 Da from C21H27N7O14P2; [M+H]+ 664.1164, [M-H]- 662.1018; deviation <= 10 ppm Pass |
| Purity | HPLC, C18 polar-endcapped, 20 mM ammonium acetate pH 5.0 and methanol, 260 nm, area % | 99.12 ± 0.20 % (k = 2) | >= 99.0 % Pass |
| Related substances | HPLC at 260 nm and 340 nm; LOD 0.01 %, LOQ 0.03 %, reporting threshold 0.05 % | alpha-NAD+ 0.32 % at RRT 0.95 (± 0.03 %); NADH 0.14 % at RRT 1.06 (340 nm positive); total impurities 0.88 %, i.e. 4.4 mg per vial | alpha-anomer <= 1.0 %, NADH <= 0.5 %, any other <= 0.5 %, total <= 2.0 % Pass |
| Impurity profile | HPLC with LC-HRMS assignment | RRT 0.84, 0.10 %, m/z 348.070 (AMP); RRT 0.89, 0.14 %, m/z 560.079 (ADP-ribose, loss of nicotinamide); RRT 0.95, 0.32 %, same mass (alpha-NAD+, anomer, enzymatically inactive); RRT 1.06, 0.14 %, m/z 666.132 (NADH); RRT 1.18, 0.18 %, m/z 123.055 (free nicotinamide) | Peaks above 0.05 % reported |
| Assay, total nucleotides | UV at 260 nm, ε 17,800 l/(mol·cm) at pH 7.0, against USP beta-NAD RS | 95.60 ± 1.0 % w/w (k = 2) | Report only |
| Assay, beta-NAD+ | UV assay × chromatographic purity; confirmed enzymatically (alcohol dehydrogenase, A340, ε 6,220) | 94.80 ± 1.1 % w/w (enzymatic 94.90 ± 1.5 % w/w; the 0.1 % agreement confirms that alpha-anomer and NADH are small) | >= 93 % w/w Pass |
| Fill weight | Gravimetric, two vials, weighed quickly (hygroscopic) | 514.2 ± 0.1 mg (second vial 512.8 mg, difference 0.3 %) | Labelled 500 mg Report only |
| beta-NAD+ per vial | Calculated from fill weight and beta-NAD+ assay | 487 mg per vial labelled 500 mg, i.e. 97.5 % of label | 95 to 105 % of label Pass |
| Form | ICP-OES (Na, K, Li); ion chromatography (phosphate) | Free acid: sodium 0.03 % w/w (a mono-sodium salt would hold 3.4 %), potassium and lithium < 0.01 %; free phosphate 0.08 % (hydrolysis product of the pyrophosphate) | Free acid; phosphate <= 0.3 % Pass |
| Trifluoroacetate and anions | Ion chromatography, LOQ 20 ppm | Trifluoroacetate < 20 ppm (not used in fermentation-derived NAD+); chloride 35 ppm; sulfate and acetate < 20 ppm; free pyrophosphate < 0.05 % | Chloride <= 500 ppm Pass |
| Water content | Coulometric Karl Fischer, USP <921>, weighed in a dry box | 3.40 ± 0.17 % w/w (k = 2), i.e. 17.5 mg per vial | <= 8.0 % Pass |
| Mass balance | beta-NAD+ + organic impurities + water + phosphate + sodium | 94.80 + 0.85 + 3.40 + 0.08 + 0.03 = 99.16 % (± 1.2 %) | 98 to 102 % Pass |
| pH | Potentiometric, 10 mg/ml in water | 3.2 (free acid; above pH 7 NAD+ hydrolyses to ADP-ribose and nicotinamide, so the acid form favours storage) | 2.5 to 3.5 Pass |
| Solubility | Visual, 50 mg/ml and 10 mg/ml | >= 50 mg/ml in water and in bacteriostatic water; clear, colourless, free of particles within 60 s; clear at 100 mg/ml, pH 2.9 | Clear solution Pass |
| Bacterial endotoxins | Kinetic chromogenic LAL, USP <85>, after adjustment to pH 6.5 to 7.5 with Tris (an acidic sample would inhibit the lysate) | < 0.005 EU/mg (< 2.5 EU per vial; LOQ 0.001 EU/mg) | <= 0.02 EU/mg Pass |
| Sterility | Membrane filtration, USP <71>; fluid thioglycollate 30 to 35 °C and soya bean casein digest 20 to 25 °C | No growth after 14 days, 2 vials | No growth Pass |
| Microbial enumeration | Plate count, USP <61>, sample neutralised to pH 7; specified organisms, USP <62> | TAMC < 10 CFU/g, TYMC < 10 CFU/g; Escherichia coli, Salmonella, Staphylococcus aureus and Pseudomonas aeruginosa absent in 1 g | TAMC <= 1000 CFU/g, TYMC <= 100 CFU/g; specified organisms absent Pass |
| Residual solvents | Headspace GC, USP <467> | Ethanol 210 ppm (precipitation from the fermentation lysate); acetone 45 ppm; methanol, acetonitrile and isopropanol each < 10 ppm (LOQ 10 ppm) | ICH Q3C: ethanol <= 5000, acetone <= 5000, methanol <= 3000, acetonitrile <= 410 ppm Pass |
| Elemental impurities | ICP-MS after microwave digestion (HNO3/H2O2), USP <233> | Cd, Pb, As, Hg < 0.05 µg/g; Co, V < 0.10 µg/g; Ni 0.12 µg/g (quantified); every result at or below 0.3 % of its limit | ICH Q3D(R2) Class 1 and 2A, parenteral PDE per 500 mg daily: Cd 4, Pb 10, As 30, Hg 6, Co 10, V 20, Ni 40 µg/g Pass |
| Anomeric and redox purity | HPLC 260 nm (alpha-NAD+) and 340 nm (NADH) | alpha-NAD+ 0.32 % (enzymatically inactive; not distinguished by MS or UV, only by chromatography and enzyme); NADH 0.14 %; A260/A340 of the whole sample > 100 | alpha-NAD+ <= 1.0 %; NADH <= 0.5 % Pass |
| Compound screening | LC-HRMS full scan m/z 100 to 1000, both polarities, against reference standards | NMN, nicotinamide riboside, nicotinic acid, NADP+, ADP, ATP, adenosine, FAD and fermentation residues (glucose, amino acids): none above 0.05 % of area beyond the impurities listed above | Nothing above 0.05 % Pass |
02The sample and the run
| Blank | Solvent blank (mobile phase A) injected before the sequence; no peak above 0.02 % of the main peak within RRT 0.80 to 1.30 (maximum 0.002 %) |
| Reference standard | USP beta-Nicotinamide Adenine Dinucleotide RS (primary) injected at the start and end of the sequence; area difference 0.3 %, retention time within 0.1 min; control sample from the previous batch within 0.2 % of its value |
03Check your copy of the file
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A copy that has been re-saved, printed to a new file or passed through a converter will not match, because each of those produces a different file. Compare the certificate exactly as it was sent to you, or the copy downloadable from this page.
fb096f5121f80ba75cd5aece795d291b59b1bff4c7082461857a945bab8a20bb
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