Validasi Metode Analisis
Metode validasi adalah proses yang digunakan untuk mengkonfirmasi bahwa prosedur analitis digunakan untuk tes tertentu cocok untuk digunakan. Hasil dari validasi metode dapat digunakan untuk menilai kualitas, keandalan dan konsistensi dari hasil analisis, melainkan merupakan bagian integral dari setiap praktek analitis yang baik.
metode Analytical perlu divalidasi atau revalidated
a. sebelum diperkenalkan ke dalam penggunaan rutin;
b. setiap kali perubahan kondisi untuk metode mana yang telah divalidasi
(misalnya, instrumen dengan karakteristik yang berbeda atau sampel yang berbeda dengan matriks)
c. kapan metode ini berubah dan perubahan di luar lingkup asli metode ini.
Metode validasi telah menerima perhatian yang cukup besar dalam literatur dan dari komite industri dan badan pengatur.
a. The US FDA cGMP permintaan di bagian metode yang akan divalidasi: Ketelitian, sensitivitas, spesifisitas, dan reproduksibilitas metode pengujian yang digunakan oleh perusahaan harus dibuat dan didokumentasikan. validasi semacam itu dan dokumentasi dapat dicapai sesuai persyaratan-persyaratan termasuk pernyataan dari setiap metode yang digunakan dalam pengujian sampel yang tepat untuk memenuhi standar akurasi dan reliabilitas, sebagaimana yang diterapkan untuk produk diuji. FDA Amerika juga
mengusulkan bimbingan industri untuk Analitik Prosedur dan Metode Validasi.
b. ISO / IEC 17025 mencakup sebuah bab tentang validasi metode dengan daftar sembilan parameter validasi. The ICH telah mengembangkan teks validasi konsensus mengenai prosedur analitis. Dokumen tersebut mencakup definisi untuk delapan karakteristik validasi. ICH juga mengembangkan pedoman dengan metodologi rinci .
c. US EPA dipersiapkan untuk menjadi pedoman pembangunan dan validasi
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AOAC, EPA dan organisasi ilmiah lain memberikan metode yang disahkan melalui studi multi-laboratorium.
USP telah menerbitkan pedoman khusus untuk validasi metode untuk evaluasi senyawa. USP mendefinisikan delapan langkah untuk validasi:
a. Ketepatan
b. Kete litia n
c. Kek hu susa n
d. Batas deteksi
e. Batas kuantisasi
f. Linieritas dan jangkauan
g. Keka sar a n
h. Ke segar a n
FDA juga telah menerbitkan petunjuk untuk validasi metode bioanalytical. Bioekuivalensi dan
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Wegscheider telah menerbitkan prosedur untuk validasi metode dengan fokus
Pelajaran ini memberikan review dan strategi untuk validasi metode analisis
Validitas dari metode spesifik harus ditunjukkan dalam percobaan laboratorium
sebaiknya ditulis dalam instruksi langkah demi langkah
1. Mengembangkan protokol validasi, sebuah prosedur operasi atau rencana validasi master untuk validasi
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5. Tentukan parameter kinerja dan kriteria penerimaan 6. Tentukan validasi eksperimen
7. Verifikasi karakteristik kinerja peralatan yang relevan
8. Kualifikasi bahan, misalnya standar dan reagen untuk kemurnian, jumlah yang cukup akurat dan stabilitas
14. Menetapkan jenis dan frekuensi uji kesesuaian sistem dan / atau pengendalian mutu analitis (AQC) cek untuk rutin
15. Dokumen validasi eksperimen dan hasil dalam laporan validasi
Keberhasilan penerimaan parameter validasi dan kriteria kinerja, oleh semua
Ruang lingkup metode dan kriteria validasi yang harus didefinisikan pada awal proses. Ini termasuk pertanyaan-pertanyaan berikut:
a. Apa analit harus terdeteksi?
b. Apa tingkat konsentrasi yang diharapkan?
c. Apakah matriks sampel?
d. Apakah ada zat campuran yang diharapkan, dan jika demikian, zat campuran tersebut harus dideteksi dan dihitung?
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i. Apa yang presisi dan akurasi diharapkan?
j. Bagaimana metode yang sesuai?
k. Jenis peralatan yang harus digunakan? Apakah metode untuk satu instrumen
tertentu, atau seharusnya itu digunakan oleh semua instrumen dari jenis yang sama?
l. Apakah metode yang digunakan dalam satu laboratorium khusus atau
seharusnya itu bisa diterapkan di semua laboratorium di satu sisi atau seluruh dunia?
m. Keterampilan apa yang telah diantisipasi pengguna metode ini?
Karakteristik performa Metode harus didasarkan pada tujuan penggunaan
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kuantitatif kuantitatif kualitatif kualitatif
batas deteksi tidak Tidak Ya tidak
batas kuantisasi
tidak Ya Tidak ya
linieritas ya Ya Tidak ya
jarak ya Ya Tidak tidak
ketelitian ya Ya Tidak ya
ketepatan ya Ya Tidak ya
kekhususan ya Ya Ya ya
kekasaran ya Ya Tidak mungkin
Tabel 2. Validasi parameter untuk tugas-tugas tertentu
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1. tersedia dalam jumlah yang cukup,
Setiap bahan lainnya dan bahan habis pakai, misalnya, kromatografi kolom, harus baru dan memenuhi syarat untuk memenuhi kriteria performa kolom. Hal ini
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5. Selektivitas dengan sampel nyata
Percobaan yang memakan waktu lebih, seperti akurasi dan kekerasan, termasuk
Selama validasi metode, parameter batas penerimaan, dan frekuensi uji
Setelah metode ini telah dikembangkan dan divalidasi, laporan validasi yang harus disiapkan adalah sebagai berikut:
a. Tujuan dan ruang lingkup metode (diterapkan, tipe).
b. Ringkasan metodologi.
c. Jenis senyawa dan matriks.
d. Semua bahan kimia, reagen, standar acuan, QC sampel dengan kemurnian, kelas, sumber atau petunjuk rinci tentang persiapan mereka.
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k. Kondisi rinci tentang bagaimana percobaan dilakukan, termasuk persiapan
sampel. Laporan tersebut harus cukup rinci untuk memastikan bahwa itu bisa direproduksi oleh teknisi kompeten dengan peralatan yang sebanding.
l. Prosedur dan perhitungan statistik representatif.
m. Prosedur QC dalam analisis rutin, misalnya, sistem tes kesesuaian.
n. Perwakilan plot, misalnya, kromatogram, spektra dan kurva kalibrasi.
o. Metode batas kinerja penerimaan data.
p. Ketidakpastian yang diharapkan dari hasil pengukuran.
s. Referensi (jika ada).
t. Ringkasan dan kesimpulan.
u. Persetujuan dengan nama, judul, tanggal dan tanda tangan dari mereka yang
bertanggung jawab untuk meninjau dan persetujuan dari prosedur tes analitis. Verifikasi Metode Standar
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persyaratan bahwa tingkat validasi metode tertentu adalah cukup untuk tujuan yang
1. Metode standar's validasi data yang memadai dan cukup untuk memenuhi persyaratan's metode laboratorium.
2. laboratorium harus dapat mencocokkan data performa seperti yang dijelaskan dalam standar.
Hasil harus didokumentasikan dalam protokol validasi. Kedua dokumen akan menjadi sumber utama untuk laporan validasi.
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Setiap kali ada perubahan yang mungkin memerlukan revalidation sebagian
a. sampel baru dengan senyawa baru atau matriks baru,
b. analis baru dengan keahlian yang berbeda,
c. instrumen baru dengan karakteristik yang berbeda,
d. Lokasi baru dengan kondisi lingkungan yang berbeda,
e. baru bahan kimia dan / atau referensi standar dan
f. modifikasi parameter analitik.
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Istilah selektivitas dan spesifisitas sering digunakan secara bergantian. Sebuah
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Prinsip-array deteksi dioda dalam HPLC dan aplikasi mereka dan batasan yang
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Ketepatan metode (Tabel 4) adalah sejauh mana hasil uji individu suntikan
The ICH membutuhkan presisi dari setidaknya 6 ulangan yang akan diukur pada
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Cancel Anytime. 100 1 100% 1,3 10 10-1 10% 2,8 1 10-2 1% 2,7 0,1 10-3 0,1% 3,7 0,01 10-4 100 ppm 5,3 0,001 10-5 10 ppm 7,3 0,0001 10-6 1 ppm 11 0,00001 10-7 100 ppb 15 0.000001 10-8 10 ppb 21 0.0000001 10-9 1 ppb 30
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b. Operator dengan pengalaman yang berbeda dan ketelitian
c. Peralatan dengan karakteristik yang berbeda, misalnya volume penundaan sistem HPLC
d. Variasi material dan kondisi instrumen, misalnya dalam HPLC, mobile komposisi fase, pH, laju alir fasa gerak
e. Variasi rincian eksperimental tidak ditentukan oleh metode
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faktor pemulihan dihitung untuk konsentrasi yang berbeda dapat digunakan untuk memperbaiki hasil akhir.
Konsentrasi harus mencakup rentang perhatian dan harus mencakup konsentrasi
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sumbu y dan konsentrasi yang sesuai pada sumbu x-, pada skala log. Garis horizontal
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Jarak
Kisaran suatu metode analisis adalah interval antara bagian atas dan tingkat bawah (termasuk tingkat-tingkat) yang telah menunjukkan akan ditentukan dengan
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konsentrasi atau massa analit uji. Dalam istilah praktis, sensitivitas kemiringan kurva
Pada kromatografi, batas deteksi adalah jumlah injeksi yang menghasilkan puncak
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Kekasaran
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otomatis).Dalam keadaan ini, pengembangan metode harus menyelidiki stabilitas analit dan standar.
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Parameter yang Harus Ada dalam Validasi Metode
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Cancel Anytime. Tugas Analitik Assay Kategori Cat 2 kuantitatif Cat 3 kualitatif Assay Cate 3
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