// THE 7X PROTOCOL

Seven checks. One traceable answer.

A fixed, finished-vial panel — no tiers, no add-ons. Every sample receives all seven checks. Every result identifies the sample, method, specification source, quality status, and technical reviewer.

LAYER
01

Homogeneity Profiling

RP-HPLC 214nm (USP <621>)
QUESTION
What proportion of detected chromatographic area is the primary component?

RP-HPLC at 214nm isolates the main peak from related substances at the peptide-bond absorbance wavelength.

METHOD STATUS: VALIDATED IN HOUSE
LAYER
02

Molecular Mass Fingerprinting

LC-MS/MS (ESI+)
QUESTION
Is the expected compound present?

LC-MS/MS confirms exact monoisotopic mass and fragmentation identity, ±1 Da. Identity consistent with expected mass.

METHOD STATUS: VALIDATED IN HOUSE
LAYER
03

Net Peptide Mass Quantitation

Quantitative HPLC vs reference std
QUESTION
How much target peptide is present per vial?

Quantitative HPLC against a qualified reference standard — stripping moisture, salts, and counter-ions like TFA. Report as mg per vial.

METHOD STATUS: VALIDATED IN HOUSE
LAYER
04

Bacterial Endotoxin

Kinetic chromogenic LAL
QUESTION
What endotoxin activity was measured?

Kinetic chromogenic LAL (USP <85>). Reported in EU/mL and EU/mg against a stated limit.

METHOD STATUS: VERIFIED COMPENDIAL
LAYER
05

Elemental Catalytic Sweep

ICP-MS (USP <232>/<233>)
QUESTION
Were specified elements detected, and at what levels?

ICP-MS (USP <232>/<233>) for Pb, As, Cd, Hg at sub-ppb. Each element: result, unit, LOQ, specification source.

METHOD STATUS: VERIFIED COMPENDIAL
LAYER
06

Microbial Quality

Rapid molecular microarray screen
QUESTION
Were viable microorganisms detected?

Rapid molecular microarray screen. NOT a USP <71> sterility test. Reported as CFU against a limit.

METHOD STATUS: RESEARCH SCREEN
LAYER
07

Post-Reconstitution Stability

Forced-degradation stability
QUESTION
Are separately prepared vials consistent, and is the reconstituted solution stable?

≥3 independent vials: individual results, mean, SD, CV. Post-reconstitution pH, clarity, and 24h degradation.

METHOD STATUS: RESEARCH SCREEN